KR20120139222A - Composition for skin external application comprising tanshinone iia as the active ingredient - Google Patents

Abstract

The present invention provides a topical skin composition containing tanshinone IIA, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient. The composition of the present invention has the effect of skin whitening, skin moisturizing, skin color improvement and skin transparency improvement.

Description

Skin external preparation composition containing tanshinone ⅡA as an active ingredient {COMPOSITION FOR SKIN EXTERNAL APPLICATION COMPRISING TANSHINONE IIA AS THE ACTIVE INGREDIENT}

The present invention relates to an external preparation composition for skin containing tanshinone IIA as an active ingredient.

Human skin is changed by a number of internal and external factors as it ages. That is, internally, the secretion of various hormones that regulate metabolism decreases, and the function of immune cells and the activity of cells decreases, thereby reducing the biosynthesis of immune proteins and constituent proteins necessary for living organisms. Due to the increase in the amount of ultraviolet rays reaching the surface of the sun's rays and further increase environmental pollution, free radicals and free radicals, such as free radicals, skin thickness, wrinkles, elasticity is reduced only In addition, skin problems frequently occur, causing a variety of changes, including increasing blemishes, freckles, and blotch. In addition, as the skin ages, capillaries are reduced or deformed, resulting in poor skin circulation, resulting in dull skin color, and rough skin.

Many factors are involved in determining the skin color of humans, including the activity of melanocytes that make melanin, the distribution of blood vessels, the thickness of skin and the presence or absence of pigments in the body, such as carotenoids and bilirubin. It is important. Especially, the most important factor is a black pigment called melanin which is produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of melanin pigment affects genetic factors, hormonal secretion, physiological factors such as stress, and environmental factors such as ultraviolet irradiation. The melanin pigment produced in melanocytes of body skin is a phenolic polymer substance having a complex form of black pigment and protein. It protects the subcutaneous skin organs by blocking ultraviolet rays irradiated from the sun, and at the same time, free radicals And protects proteins and genes in the skin. As described above, melanin produced by stress stimulation inside and outside the skin is a stable substance that does not disappear until it is discharged to the outside through skin keratinization even if the stress disappears. However, if more melanin is produced than necessary, it causes hyperpigmentation such as blemishes, freckles, and spots, resulting in cosmetically bad results. Today, Asian women prefer white and clean skin like white jade, and this is an important standard of beauty, increasing the demand for treatment and cosmetic problems for hyperpigmentation. In response to this demand, ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione or derivatives thereof, or substances having tyrosinase inhibitory activity have been used in combination with cosmetics or pharmaceuticals, but they are insufficient. Its use has been limited due to its whitening effect, safety issues on the skin, formulation and stability problems when formulated in cosmetics, and the like.

The most important function of the outermost epidermis of the skin is to protect the skin from various external stimuli (chemicals, air pollutants, dry environment, physical and chemical stimulating factors such as ultraviolet rays). It is a protective function that prevents excessive divergence of moisture through the body, and this protective function can be maintained only when the stratum corneum composed of keratinocytes is normally formed. The outermost stratum corneum (horney layer) of the epidermis is formed from keratinocytes and is composed of the differentiated keratinocytes and the surrounding lipid layer. Keratinocytes are characteristic cells whose basal cells, which continuously proliferate in the epidermal layer, undergo morphological and functional changes in stages and rise to the surface of the skin. Exfoliated and new keratinocytes take over their functions. This repetitive sequence of changes is called "epidermal differentiation" or "keratinization." During the keratinization process, keratinocytes form the stratum corneum by generating natural moisturizing factor (NMF) and intercellular fat (ceramide, cholesterol, fatty acid), making the stratum corneum firm and flexible. It has a function as).

However, these stratum corneum can easily lose its function due to excessive face washing, lifestyle factors such as bathing, environmental factors such as dry air and pollutants, and endogenous diseases such as atopic or aged skin. In fact, due to a number of factors that have increased in modern times, more and more people are complaining of dry skin symptoms and the resulting disability. Therefore, many studies have been conducted to supply moisture from the outside or to prevent moisture loss from the body in order to maintain proper skin moisture, and various types of moisturizers having moisture retention capabilities have been developed and mainly cosmetics. It is widely used in the field. However, as the risk factors for humans increase and the aged population rapidly increases in the living environment, the turnover rate of the stratum corneum is slowed, the lipid synthesis ability of the keratinocytes is degraded, or the epidermis is normal in the epidermis. Due to poor cell division, maturation and differentiation, the amount of moisturizing factors and lipids in the stratum corneum is reduced, so that humans with skin with a condition that is unable to maintain normal stratum corneum function, that is, the skin barrier function is degraded There is a growing trend. Due to the abnormal division and differentiation of epidermal cells, the skin causes various skin diseases such as dry skin, atopy and psoriasis, and these diseases can alleviate the symptoms slightly by using a conventional moisturizer having only water retention function. However, it is difficult to expect radical healing.

The present invention aims to provide a skin external preparation composition having a skin whitening effect, a skin moisturizing effect, a skin color improving effect, and a skin clarifying effect while being able to safely use without side effects on the skin to solve the above problems.

In order to achieve the above object, an embodiment of the present invention is an external preparation composition for skin containing tanshinone IIA represented by the following formula (1), an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient. To provide.

In one embodiment of the present invention, the content of the tanshinone ⅡA may be from 0.0001 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the composition may be an external skin composition for skin whitening having an effect of inhibiting melanin production.

In one embodiment of the present invention, the composition may be a skin external preparation composition for moisturizing the skin having an effect of promoting differentiation of keratinocytes.

In one embodiment of the present invention, the composition may be a skin external preparation composition for moisturizing the skin having the effect of increasing the expression of transglutaminase (transglutaminase) of the skin cell line.

In one embodiment of the present invention, the composition may be an external skin composition for improving skin color and improving skin transparency having an effect of increasing NO production in vascular endothelial cells.

In one embodiment of the present invention, the composition may be a skin external preparation composition for improving skin color and improving skin clarity having an effect of improving peripheral blood circulation.

In one embodiment of the invention the composition may be a pharmaceutical composition.

In one embodiment of the present invention, the pharmaceutical composition may be a formulation selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions and emulsions.

In one embodiment of the present invention, the composition may be a cosmetic composition.

In one embodiment of the present invention, the cosmetic composition is a softening lotion, nourishing lotion, lotion, body lotion, nutrition cream, massage cream, moisturizing cream, hand cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, It may be a formulation selected from the group consisting of gels, patches, lipsticks, makeup bases and foundations.

The composition of the present invention may have a skin whitening effect, a skin moisturizing effect, a skin color improving effect, and a skin transparency improving effect.

Hereinafter, the present invention will be described in detail.

One embodiment of the present invention provides a topical skin composition containing tanshinone IIA, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient.

Said "isomers" include in particular optical isomers, form isomers, positional isomers (particularly tautomers) or geometric isomers.

The precursor refers to a chemical change of a compound to control physical and chemical properties. The precursor itself does not exhibit physiological activity, but may be converted to the original compound by the action of chemical or enzymes in the body after administration, thereby exerting an effect. .

The salts refer to pharmaceutically acceptable salts, wherein the "pharmaceutically acceptable" is generally useful for preparing safe, nontoxic, biologically or otherwise desirable pharmaceutical compositions, and is useful for humans as well as for veterinary use. It is also useful for pharmaceutical use.

The term "pharmaceutically acceptable salts" refers to salts that are pharmaceutically acceptable and have the desired pharmacological activity as defined above. Such salts include (1) inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, stone Sinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2- Formed by en-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphric acid, salicylic acid, stearic acid, muconic acid and the like Acid addition salts; Or (2) salts formed when the acidic protons present in the parent compound are replaced.

The term “hydrate” means “pharmaceutically acceptable hydrate”. The "hydrate" is present when the compound of the present invention contains water. Hydrates may contain one or more water molecules per molecule of the compounds of the invention. Exemplary non-limiting examples include monohydrates, dihydrates, trihydrates, and tetrahydrates. Hydrates may contain one or more compound molecules of the invention per one water molecule. Exemplary non-limiting examples include semihydrates. In one embodiment, water may be retained in the crystal in a variety of ways such that water molecules may occupy lattice positions within the crystal, or may form bonds with salts of the compounds described above. A luggage must be "acceptable" in the sense that it is not harmful to its reception.

The term "solvate" means "pharmaceutically acceptable solvate" and "solvate" means that the compound of the present invention contains one or more pharmaceutically acceptable solvents. The solvate may contain one or more solvent molecules for one molecule of the compound of the present invention, or may contain one molecule of the compound of the present invention for one molecule of the solvent. In one embodiment, the solvent may be retained in the crystal in various ways such that water molecules may occupy a lattice position in the crystal, or may form bonds with salts of the compounds described above.

Tanshinone IIA may be represented by the following [Formula 1].

 [Formula 1]

The content of the tanshinone ⅡA may be 0.0001 to 10% by weight based on the total weight of the composition. When the content is less than 0.0001% by weight, the efficacy is weak, and when it exceeds 10% by weight, there is no apparent increase in efficacy due to the increase in content, which is not economical.

The tanshinone ⅡA can also be produced by organic synthesis, or can be obtained from the extract of Salvia.

Dansam is a perennial herbaceous plant of Lamiaceae (Sun-Hyun, Labiatae), which has tinnitus such as red ginseng, powdered horseweed, and sheep oil. Root stocks are short and coarse and usually have stem marks at their ends. Roots are long cylindrical, divided into several, the outer side is reddish brown or dark reddish brown with rough and vertical wrinkles. Salvia mimics blood circulation, removes blood, and relieves limb pain. Pharmacological actions include increased cardiovascular blood flow, decreased serum lipids, improved microcirculation, antithrombotic action, sedative action, antimicrobial action, immune enhancement, anticancer action, and hypoglycemic effect. For example, the Salvia Militija bungee ( Salvia) miltiorrhiza Bunge ), and Salvia extract can be obtained from the root, for example.

The extract as defined in the present invention is characterized in that the crude extract, polar solvent soluble extract or non-polar solvent soluble extract. The crude extract includes water containing purified water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably a water and ethanol mixed solvent, and more preferably 75 to 100% ethanol extract.

The polar solvent soluble extract is an extract selected from water, methanol, butanol or a mixed solvent thereof, preferably n-butanol, and the non-polar solvent soluble extract is hexane, chloroform, methylene chloride, ethyl acetate, glycerin, Extracts soluble in propylene glycol, butylene glycol or ether, preferably methylene chloride or ethyl acetate.

The extract is more preferably, after primary extraction using any one or more of a polar solvent containing anhydrous and hydrous lower alcohols having 1 to 4 carbon atoms, acetone and butylene glycol, and then sequentially ethyl acetate, diethyl acetate Extraction may be performed through secondary extraction using any one or more of a low polar solvent including diethyl ether, benzene, chloroform and hexane. More specifically, as an extraction solvent, water, anhydrous and hydrous lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, normal propanol, normal butanol), a mixture of water and lower alcohols, acetone, 1,3-butylene glycol, isopropanol 5 to 20 times the volume of the polar solvent and their mixed solvent to be added, based on the dry weight of the pulverized product of the red ginseng root, immersion extraction and filtration, and the filtrate is concentrated under reduced pressure. Water is added to the resulting concentrate to disperse the mixture, followed by shaking with one or more of a low polar organic solvent containing the same amount of ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane. Thereafter, the organic layer is separated and concentrated under reduced pressure to obtain a root extract of Salvia.

Extraction method is carried out for about 12 to 96 hours in the case of cold needle, or left for 3 to 20 days at room temperature of 4 to 25 ℃ using an anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, ethyl acetate or diethyl ether as a solvent The active ingredient can be extracted by aging. In the case of warm needle, it is different depending on the type and temperature of the extraction solvent, but it is preferably performed at a temperature close to the reflux temperature of the solvent for about 5 to 24 hours.

By the extraction method to extract sequentially according to the polarity change as described above, it is possible to obtain the extract of maximizing the active ingredient and removing impurities to improve stability and at the same time maximize the efficacy.

In addition, the extraction is subjected to filtration, in the present invention, the filtration is a process of removing the suspended solid particles from the extract, by filtering the particles using cotton, nylon, or the like, ultrafiltration, cryofiltration, centrifugation, etc. This is not restrictive.

It may also further comprise a separation process by various chromatography (chromatography according to size, charge, hydrophobicity or affinity).

Next, the step of drying the filtrate includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, infrared drying and the like. If desired, a process of grinding the final dried extract may be added.

The external preparation composition for skin according to the present invention generally is a concept encompassing the overall composition used for external application of the skin, and may be, for example, a pharmaceutical composition or a cosmetic composition.

In one embodiment of the present invention, the pharmaceutical composition may be formulated as a parenteral dosage form in solid, semi-solid or liquid form by adding a commercially available inorganic or organic carrier, excipient and diluent with tanshinone IIA as an active ingredient. For parenteral administration, the preparation may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions and emulsions, but is not limited thereto.

Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the external composition for skin according to each formulation, other ingredients other than the composition of the present invention described above may be appropriately selected and blended by those skilled in the art according to the formulation or use purpose of the external skin preparation, and in this case, simultaneously applied with other raw materials. If you do, synergistic effects may occur.

In addition, when the composition according to the present invention is used as a pharmaceutical composition, it may further contain pharmaceutical supplements and other therapeutically useful substances such as preservatives, stabilizers, hydrating or emulsifiers, salts or buffers for controlling osmotic pressure. And may be formulated in parenteral dosage forms according to conventional methods.

It is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the severity of the symptom, the route of administration chosen, the age, sex, weight and health of the subject.

In general, the dosage of the active ingredient ranges from 0.001 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 0.5 mg / kg / day to 2.5 mg / kg / day. Topical administration may be administered once a day or may be divided several times.

In one embodiment of the present invention, the cosmetic composition may include, for example, a basic cosmetic, makeup cosmetics, cosmetics for the body, the formulation is not particularly limited and may be appropriately selected as desired.

For example, the cosmetic composition is formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, sprays, and the like. It may be, but is not limited thereto. More specifically, softening cream, nourishing cream, lotion, body lotion, nourishing cream, massage cream, moisturizing cream, hand cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, gel, patch, oil in water It may be formulated into a cosmetic composition such as (O / W) type, basic cosmetics such as water-in-oil (O / W) type, color cosmetics such as lipstick, makeup base or foundation, shampoo, rinse, body cleanser and the like.

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

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

In the case where 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 a paste, cream or gel, an animal oil, vegetable oil, 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.

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, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

In one embodiment of the present invention, the cosmetic composition may further contain a thickener. The thickener included in the cosmetic composition of the present invention is methyl cellulose, carboxy methyl cellulose, carboxy methyl hydroxy guanine, hydroxy methyl cellulose, hydroxyethyl cellulose, carboxy vinyl polymer, polyquaternium, cetearyl alcohol, stearic acid, Carrageenan and the like can be used, and preferably, at least one of carboxy methyl cellulose, carboxy vinyl polymer and polyquaternium can be used, and most preferably, carboxy vinyl polymer.

In one embodiment of the present invention, the cosmetic composition may contain a variety of suitable bases and additives as necessary, the type and amount of these components can be easily selected by the inventors. If necessary, it may contain an acceptable additive, for example, may further include components such as preservatives, pigments, additives and the like conventional in the art.

The preservative may specifically be phenoxyethanol or 1,2-hexanediol, and the perfume may be an artificial perfume or the like.

In addition, in one embodiment of the present invention, the cosmetic composition may include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts. Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, purified water and the like.

In addition, the compounding component which may be added other than this is not limited to this, Moreover, any of the above components can be mix | blended within the range which does not impair the objective and effect of this invention.

Hereinafter, the present invention will be described in detail with reference to Examples, Comparative Examples and Test Examples of the present invention. These are only presented by way of example only to more specifically describe the present invention, it is to those skilled in the art that the scope of the present invention is not limited by these Examples, Comparative Examples and Test Examples. Will be self explanatory.

Example 1 Preparation of Salvia Root Extract

Salvia roots were washed with purified water, dried and triturated. 200 g of granulated salvia root powder was added to 1 liter of an aqueous 70% ethanol solution, and extracted by boiling in an extractor equipped with a cooling condenser for 12 hours, followed by filtration with a 300 mesh filter cloth. The filtrate was left to mature at 4-15 ° C. for 7 days and then filtered through Whatman No. 2 filter paper. Thereafter, the extract was added to a 3-liter separatory funnel, and 1 liter of ethyl acetate was added thereto, shaken well, the mixture was thoroughly separated, and the upper layer (ethyl acetate layer) was taken. The lower layer (water layer) was again extracted twice with a separatory funnel. The combined upper layers were combined, concentrated under reduced pressure at 50 ° C. using a distillation apparatus equipped with a cooling condenser, and dried to obtain a dry weight 45.5 g of Dansam root extract.

Example 2 Isolation of Tanshinone IIA from Salvia Radix Extract

20 g of Salvia Militiorrhiza root extract obtained using the method of Example 1 was placed on a silica gel column (70-230 mesh, Merck 7734), and hexane (Hexane)-> hexane-methyl chloride (Hexane-CH ₂ Cl ₂ (1: 1) Tanninone IIA was isolated by gradient elution with)), which was then recrystallized with EtOAc to give 43.2 mg of a yellowish red needle. Table 1 below shows NMR data of tanshinone IIA.

Position ¹ H-NMR (500 MHz, CDCl ₃ ) ¹³ C-NMR (500 MHz, CDCl ₃ ) One 3.19 ( t , 6.5) 29.89 ( t ) 2 1.79 ( m ) 19.13 ( t ) 3 1.66 ( m ) 37.87 ( t ) 4 - 34.67 ( s ) 5 - 150.14 ( s ) 6 7.63 ( d , 8.0) 133.47 ( d ) 7 7.55 ( d , 8.0) 119.91 ( d ) 8 X 127.48 ( s ) 9 X 126.52 ( s ) 10 X 144.48 ( s ) 11 - 183.66 ( s ) 12 - 175.99 ( s ) 13 - 121.16 ( s ) 14 - 161.73 ( s ) 15 7.22 ( d , 1.0) 141.29 ( d ) 16 - 120.25 ( s ) 17 2.26 ( s ) 8.80 ( q ) 18 1.31 ( s ) 31.80 ( q ) 19 1.31 ( s ) 31.80 ( q ) 20 - -

Test Example 1 Skin Whitening Efficacy Test

1) Determination of melanin production inhibitory effect using pigment cells in rats

To determine the melanin production inhibitory effect of tanshinone ⅡA extract was measured using the pigment cells of the mouse.

First, the mouse Mel-Ab cells derived from C57BL / 6 mice (Dooley, TP et al, Skin pharmacol, 7, pp 188-200) were placed in DMEM (Dulbeccos modified Eagles media) in 10% fetal placental serum, 100 nM 2-O-tetradecanoylphorbol-13-ate and 1 nM cholera toxin were incubated at 37 ° C. and 5% CO 2. The cultured Mel-Ab cells were separated with 0.25% trypsin-EDTA, the cells were incubated at a concentration of 105 cells / well in a 24-well plate, and 1 ppm, 10 ppm of tanshinone IIA for 3 consecutive days from the second day. The extract was added and incubated. At this time, hydroquinone was used as a control. Then, the culture medium was removed, washed with PBS, the cells were dissolved with 1N sodium hydroxide, the absorbance was measured at 400 nm, and the melanin production inhibition rate was calculated according to Equation 1 below and the results are shown in Table 2 (Dooley). Method).

Test substance Melanin production inhibition rate (%) Tancinone IIA Extract 0.0001% by weight 10.4 Tanshinone IIA Extract 0.001% by weight 32.9 Hydroquinone (control) 40.5

As shown in Table 2, the tanshinone ⅡA extract of the present invention showed an excellent melanin production inhibition rate, and showed a melanin production inhibition rate similar to hydroquinone. Through this it was found that the tanshinone ⅡA extract of the present invention has an excellent whitening effect.

2) Whitening effect test on human skin

In order to investigate the whitening effect of tanshinone ⅡA extract on the human skin, the following experiment was performed.

First, an opaque tape with a hole of 1.5 cm in diameter was attached to the upper arm of 10 healthy men, and then 1.5 ~ 2 times of ultraviolet light (UVB) of the minimum erythema dose of each subject. Was irradiated to induce skin blackening.

After the ultraviolet irradiation, tanshinone ⅡA extract and hydroquinone were applied as test materials, respectively, and one state was observed for 10 weeks without applying anything. Skin color was measured on a weekly basis with a colorimeter CR2002 (Minolta, Japan).

Then, the difference in skin color (ΔL *) between the start point of application and the end point of application of each test substance was calculated according to the following equation, which is shown in Table 3 below. On the other hand, the whitening effect is determined by comparing ΔL * between the sample application site and the control site. When ΔL * value is about 2, the whitening effect of the deposited pigment is clear, and when it is about 1.5 or more, the whitening effect is determined. Can be.

Test substance Skin color brightness degree (△ L *) Tanshinone ⅡA Extract 1.70 ± 0.20 Hydroquinone (positive control) 2.00 ± 0.16 Vehicle (negative control) 0.45 ± 0.20

As shown in Table 3, it was confirmed that the tanshinone ⅡA extract of the present invention showed a similar degree of skin color brightness as hydroquinone. This is because the tanshinone ⅡA extract brightens the skin color by improving pigmentation produced by ultraviolet light.

[Test Example 3] skin moisturizing and atopic symptoms improvement test

1) Differentiation induction test of human keratinocytes

The amount of Cornified Envelop (CE) produced during the differentiation of keratinocytes and human skin cell lines (HaCaT) was measured and tested by absorbance to see the effect of promoting the differentiation of tanshinone IIA extract.

In the test using absorbance, keratinocytes of primary culture were put in a culture flask and attached to the bottom, and then the test substances of Table 4 were added to the culture solution at the concentrations shown in Table 4, and the cells were 70% of the floor area. Incubated for 5 days until the growth of ~ 80%. The cells were harvested and washed with PBS (phosphate buffered saline), followed by 10 mM Tris-HCl (Tris-HCl) containing 2% Sodium Dodecyl Sulfate (SDS) and 20 mM Dithiothreitol (DTT). , pH 7.4) 1 ml was added, and the precipitates sonicated, boiled, and centrifuged were again suspended in 1 ml of PBS to measure absorbance at 340 nm. Separately, a portion of the solution after the sonication was taken to measure the protein content, which was used as a reference when evaluating the degree of cell differentiation. Low calcium (0.03mM) treated group and high calcium (1.2 mM) treated group as a negative / positive control, respectively, the test results performed by adding the test substance to the low calcium concentration is shown in Table 4 below.

Test substance Differentiation capacity in keratinocytes (%) Control group Low Ca ^{2 +} (0.03 mM) 100 Hign Ca ^{2 +} (1.2 mM) 198 Tanshinone ⅡA Extract 0.0001% by weight 110 0.001 wt% 122 0.01 wt% 125

As shown in Table 4, the tanshinone IIA extract according to the present invention can be seen to promote differentiation in keratinocytes. Therefore, tanshinone ⅡA extract has a skin moisturizing effect.

2) Expression effect of transglutaminase of human skin cell line

Transglutaminase is a kind of skin differentiation marker, which helps to form the formed corneal envelope (CE) produced by the differentiation of human skin cell line (HaCaT) and cell differentiation.

Human skin cell lines were placed in each well of 96-well cell culture plates and ⁴ × 10 ⁴ cells were attached for 24 hours. After treatment with the test substance to the attached skin cell line, after 2 days, the medium was removed, and stored in a -20 ℃ refrigerator. After freeze-thawing was repeated twice, the cells treated with material were destroyed, and then treated with acetone: ethanol (1: 1, v / v) stored at -20 ° C and left at 4 ° C for 30 minutes. Fix the cells. Thereafter, the organic solvent was allowed to evaporate at room temperature, blocking (1% bovine serum albumin), transglutaminase antibody, HRP anti-mouse antibody (secondary) were performed, and color development was OPD (o). -phennyldiamine) was added. Expression was measured by absorbance at 490nm, calibration was performed by measuring the background (background) at 630nm. Low calcium (0.03mM) treated group and high calcium (1.2 mM) treated group as a negative / positive control, respectively, and the test results performed by adding the test substance to the low calcium concentration is shown in Table 5 below.

Test substance Transglutaminase Expression (%) Control group Low Ca ^{2 +} (0.03 mM) 100 Hign Ca ^{2 +} (1.2 mM) 135 Tanshinone ⅡA Extract 0.0001% by weight 130 0.001 wt% 172 0.01 wt% 208

As shown in Table 5, the tanshinone ⅡA extract according to the present invention showed a two-fold increase compared to the control, it can be seen that the amount is expressed more than the control. Therefore, it was found that the tanshinone ⅡA extract according to the present invention improves the expression of transglutaminase.

3) Measurement of skin moisturizing increase in human body

40 male and female adults in their 50's and 60's showing skin dryness were divided into two groups, and Example 1 (containing tanshinone IIA extract) and Comparative Example 1 were applied to the face twice daily for 4 weeks in each group.

Table 6 shows the compositions of Example 1 and Comparative Example 1.

ingredient Example 1 Comparative Example 1 Content (% by weight) Content (% by weight) Tanshinone IIA Extract 1.0 - glycerin 5.0 5.0 Oleyl alcohol 1.5 1.5 ethanol 5.5 5.5 Polysorbate 80 3.2 3.2 Carboxyl Vinyl Polymer 0.1 0.1 Butylene Glycol 3.0 3.0 Propylene glycol 3.0 3.0 Preservative, fragrance Suitable amount Suitable amount Purified water Balance Balance system 100 100

Koniometer at constant temperature and humidity conditions (24 ° C, 40% relative humidity) before start of application, after 1 week, 2 weeks, 4 weeks after application, and after 2 weeks (total 6 weeks) after application was stopped. Skin moisture content was measured by. The results are shown in Table 7 below. The test result is a percentage of the increase of the measured value after a certain period of treatment based on the Koniometer value measured immediately before the test was started.

Test group Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Example 1
(Contains Tanshinone IIA Extract) 32 40 42 32 Comparative Example 1
Control group 30 32 33 18

The Koniometer is a skin moisture meter that measures the amount of moisture present on the skin by measuring the conductivity of the epidermis. As shown in Table 7, in the group to which the material of Example 1 containing tanshinone IIA was applied, the amount of skin moisture was further increased compared to the control to which Comparative Example 1 was applied. Also, two weeks after the application of the test substance was stopped, the value after 6 weeks after the measurement of the skin moisture was similar to that after 1 to 2 weeks. It can be seen that the skin moisture is maintained by.

Test Example 3 Skin Color Improvement Test

1) NO-producing ability of tanshinone ⅡA extract in human umbilical vein endothelial cells (HUVEC)

Endothelial nitric oxide synthase (eNOS) is present in human vascular endothelial cells, and its activity is increased to produce nitric oxide (NO) to expand blood vessels and promote blood circulation. After culturing human vascular endothelial cells, the amount of NO produced by treating tanninone IIA extract with the medium was compared. Table 8 below shows NO production by tanshinone ⅡA extract.

Test substance NO (nitric oxide) production performance (%) Control group Low Ca ^{2 +} (0.03 mM) 100 Tanshinone ⅡA Extract 0.005% by weight 320 0.01 wt% 880 0.02 wt% 1350

As can be seen in the results of Table 8, the tanshinone ⅡA extract showed excellent NO production in vascular endothelial cells. The superior nitric oxide (NO) production of tanshinone ⅡA extract in the end results in the expansion of capillaries and accelerated blood circulation, providing nutrients to the skin smoothly, inhibiting skin aging, and improving skin color. Will contribute. It also contributes to improving skin clarity.

2) Test to improve the color of human skin (face)

In order to evaluate the blood circulation promoting effect on the skin of the composition containing the tanshinone ⅡA extract of the present invention, the degree of blood circulation in the skin was measured using a Laser Doppler Perfusion Imager (LDPI). A well-known and still used device for measuring blood circulation, it is a very sensitive device that can measure not only the velocity and amount of blood in the capillaries of the skin but also the flow in the small arteries and the venous veins.

After washing the face with soap in a constant temperature and humidity room for 30 minutes, the initial values were measured using an IRPI (Infrad) camera, an LDPI and skin temperature measuring instrument. The participants were 20 women with cold hands and feet. LDPI was used to measure the initial blood flow in the lower part of the forehead, and the IR camera was used to measure the initial skin temperature of the forehead, eyes, and cheeks.

For use in the test, Example 1 having the composition shown in Table 6 above was prepared and the blood flow and skin temperature after using Example 1 for one week were compared with the initial measurements. LDPI results before and after the use of Example 1 are shown in Table 9 below, and IR results before and after use are shown in Table 10.

Face Before use After 1 week medium 0.952 1.169

forehead Under the eyes cheek Before use 32.26 32.02 30.44 After 1 week 33.87 33.69 33.57

As a result of using Example 1 containing the tanshinone ⅡA extract as described above, it was shown that the blood color is improved as a peripheral blood circulation improvement effect of the face, which ultimately effectively delivers the skin's nutrients and contributes to inhibiting and delaying skin aging. Done.

Hereinafter, the formulation examples of the cosmetic composition and the pharmaceutical composition according to the present invention will be described, but can be applied in various formulations in addition to the following examples, which are intended to explain in detail only, not intended to limit the present invention.

Formulation Example 1 Flexible Lotion (Skin Lotion)

To a flexible lotion was prepared in a conventional manner according to the composition shown in Table 11.

ingredient Content (% by weight) Tanshinone ⅡA Extract 1.0 glycerin 3.5 Oleyl alcohol 1.5 ethanol 5.5 Polysorbate 80 3.2 Carboxyl Vinyl Polymer 0.1 Butylene Glycol 2.0 Propylene glycol 2.0 Preservative, fragrance Suitable amount Purified water Balance system 100

Formulation Example 2 Nutritional Lotion (Milk Lotion)

To the nutritional lotion was prepared in a conventional manner according to the composition shown in Table 12.

ingredient content
(weight%) Tanshinone ⅡA Extract 1.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.2 Wax 4.0 Polysorbate 60 1.5 Caprylic / Capric Reglycerides 5.0 Squalane 5.0 Sorbitassquioleate 1.5 Cetearyl Alcohol 1.0 Triethanolamine 0.2 Preservative, fragrance Suitable amount Purified water Balance system 100

Formulation Example 3 Nutrition Cream

To the nutrition cream was prepared in a conventional manner according to the composition described in Table 13.

ingredient content
(weight%) Tanshinone ⅡA Extract 1.0 glycerin 3.5 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / Capric Reglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Triethanolamine 0.1 Preservative, fragrance Suitable amount Purified water Balance system 100

Formulation Example 4 Massage Cream

To prepare a massage cream in a conventional manner according to the composition described in Table 14.

ingredient Content (% by weight) Tanshinone ⅡA Extract 1.0 glycerin 8.0 Butylene glycol 3.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / Capric Triglycerides 3.0 Wax 4.0 Cetearyl Glucoside 1.5 Sesqui oleic acid sorbitan 0.9 paraffin 1.5 Preservative, coloring, fragrance Suitable amount Purified water Balance system 100

[Formulation Example 5] Pack

To prepare a pack in a conventional manner according to the composition described in Table 15 below.

ingredient Content (% by weight) Tanshinone ⅡA Extract 1.0 glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Nonyl Phenyl Ether 0.4 Polysorbate 60 1.2 Ethanol preservative Suitable amount Preservative, fragrance Suitable amount Purified water Balance system 100

Formulation Example 6 Pharmaceutical for Topical Patch Patch Administration

Patches were prepared by conventional methods according to the compositions set forth in Table 16 below.

ingredient Content (% by weight) Tanshinone ⅡA Extract 1.0 Beta-1,3-glucan 3.0 Diethylamine 0.7 Sodium sulphate 0.1 Polyoxyethylene lauryl ether (E.O = 9) 1.0 Polyhydroxyethylenecetylstearyl ether (Cetomacrogol 1000) 1.0 Viscous Paraffin Oil 2.5 Caprylic Acid Ester / Capric Acid Ester (Cetiol LC) 2.5 Polyethylene glycol 400 3.0 Polyacrylic Acid (Carbopol 934P) 1.0 Purified water Balance system 100

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

An external preparation composition for skin containing tanshinone IIA, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient. The method of claim 1,
The tanshinone ⅡA content is a skin external composition, characterized in that 0.0001 to 10% by weight based on the total weight of the composition. The method of claim 1,
The composition is a skin external composition for skin whitening having an effect of inhibiting the production of melanin. The method of claim 1,
The composition is a skin external composition for moisturizing the skin having an effect of promoting the differentiation of keratinocytes. The method of claim 1,
The composition is an external skin composition for skin moisturizing having the effect of increasing the expression of transglutaminase of the skin cell line. The method of claim 1,
The composition is an external composition for external skin, characterized in that for improving skin color and improving skin transparency having an effect of increasing NO production in vascular endothelial cells. The method of claim 1,
The composition is an external composition for external skin, characterized in that for improving skin color and improving skin clarity having an effect of improving peripheral blood circulation.