KR101481208B1 - Composition for skin external application comprising cryptotanshinone as the active ingredient - Google Patents

Abstract

The present invention provides a composition for external application for skin comprising, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof, or a solvate thereof. The composition of the present invention has effects of inhibiting skin aging, improving skin wrinkles, skin whitening, skin moisturizing, atopic improvement, skin color improvement and skin transparency improvement.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for external application for skin, which contains cryptoxanthinone as an active ingredient,

The present invention relates to a composition for external application for skin containing cryptotansinone as an active ingredient.

The human skin undergoes changes due to various internal and external factors as it gets older. In other words, internally, secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction As the amount of ultraviolet rays reaching the surface of the sunlight increases and the environmental pollution is further intensified, free radicals and active noxious oxygen are increased, thereby decreasing the thickness of the skin, increasing the wrinkles and decreasing the elasticity However, skin troubles are frequent, and spots, freckles, and black spots also increase. In addition, the capillary blood vessels are reduced or deformed due to aging of the skin, and the skin blood circulation becomes poor, and the skin color becomes dull and the skin becomes rough.

As the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid, and glycoprotein, which constitute the skin, change or decrease, and oxidative stress due to free radicals and active noxious oxygen occurs. Cox-2 (cyclooxygenase), an enzyme that produces a proinflammatory cytokine, known to cause inflammation in most cells constituting the skin by aging or ultraviolet rays, And the biosynthesis of Matrix metalloproteinase (MMP), an enzyme that degrades skin tissue, is increased by these inflammatory factors, and nitric oxide (NO) production by iNOS (inducible nitric oxide synthase) Respectively. In other words, decrease of cellular activity due to natural endogenous aging and decrease of biosynthesis of substrate material by microinflammation, increase of stress due to various harmful environment, increase of active oxygen species by sunlight, As the degradation and denaturation are accelerated, the skin substrate is destroyed and thinned, and various symptoms of skin aging appear. Therefore, it is a reality that many researches have been conducted on active ingredients that can prevent and improve such aging phenomena.

Several factors are involved in determining the skin color of a person. Among them, factors such as the activity of the melanocyte making the melanin pigment, the distribution of blood vessels, the thickness of the skin, and the presence or absence of pigment in and out of the body such as carotenoids and bilirubin It is important. Especially, the most important factor is melanin, a melanin pigment 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. Melanin, which is produced by stress stimuli inside and outside of the skin, is a stable substance that does not disappear until the skin is excreted through skin keratinization even if the stress disappears. However, when melanin is produced more than necessary, it induces hypercholesterolemia such as spots, freckles, and dots, resulting in poor cosmetic results. Today, oriental women prefer white, clean skin like white whites, and this is an important criterion of beauty, so there has been a growing demand for treatment and cosmetic problems for hyperpigmentation. In response to such demands, substances having ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione, derivatives thereof, or tyrosinase inhibiting activity have been conventionally used in cosmetics or pharmaceuticals, The use thereof is limited due to a whitening effect, a safety problem to the skin, a problem of formulation and stability in combination with cosmetics, and the like.

The most important function of the epidermis in the outermost skin is to protect against external stimuli (chemical, air pollutants, dry environment, physical and chemical stimulants such as ultraviolet rays) This protective function can maintain its function only when the stratum corneum composed of keratinocytes is normally formed. The outermost stratum corneum (horney layer) is formed from keratinocytes and consists of keratinocytes with complete differentiation and a surrounding lipid layer. The keratinocyte is a characteristic cell in which basal cells continuously proliferating in the lowest epidermis undergo a stepwise change in morphology and function and are elevated to the surface of the skin. After a certain period of time, And the new keratinocyte replaces its function. This repetitive sequence of changes is called "epidermal cell differentiation" or "keratinization". During keratinization, keratinocytes form the stratum corneum while producing natural moisturizing factors (NMF) and intercellular lipids (ceramides, cholesterol, fatty acids), which make the stratum corneum firm and flexible, As shown in Fig.

However, such a stratum corneum can be easily lost due to excessive harshness, environmental factors such as living habits such as bath, dry atmosphere, pollutants, and endogenous diseases such as atopic skin and senile skin, In fact, due to the increasing number of factors in modern times, the number of people who are suffering from dry skin symptoms and disorders has increased recently. Therefore, in order to maintain proper skin moisture, studies have been conducted to supply moisture from the outside or to prevent water loss from the body, and a variety of moisturizers having moisture retention ability have been developed, It is widely used in the area. However, as the risk factors for humans increase in the living environment and the aging population rapidly increases, the turnover rate of the stratum corneum is delayed, the lipid synthesis ability of the keratinocytes decreases, The cells are not smoothly divided, maturated and differentiated, and the amount of moisturizing factors and lipids in the stratum corneum is decreased, so that the normal stratum corneum does not maintain its function. Namely, . Due to the division and differentiation of abnormal abnormal epidermal cells, the skin causes various skin diseases such as dry skin, atopy and psoriasis. These diseases can alleviate the symptoms only by a conventional moisturizing agent having only moisture holding function , But it is difficult to expect fundamental healing.

The present invention provides a composition for external application for skin, which can be safely used without side effects on the skin and has skin aging inhibition, skin whitening effect, skin moisturizing effect, atopic improvement effect, skin color improvement effect and skin transparency effect .

In order to achieve the above object, an embodiment of the present invention provides a composition for external application for skin comprising, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof.

In one embodiment of the present invention, the content of cryptoxanthin may be 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 a composition for inhibiting skin aging and improving skin wrinkles, which has an effect of inhibiting the expression of Matrix metalloproteinase-1 (MMP-1).

In one embodiment of the present invention, the composition may be a composition for inhibiting skin aging and improving skin wrinkles, which has an effect of increasing the production of Type I procollagen (Type 1 Procollagen).

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

In one embodiment of the present invention, the composition may be an external composition for skin moisturizing and atopic dermatitis, which has an effect of promoting differentiation of keratinocytes.

In one embodiment of the present invention, the composition may be an external skin composition for skin moisturizing and atopic dermatitis, which has an effect of increasing the expression of transglutaminase of a skin cell line.

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

In one embodiment of the present invention, the composition may be a composition for improving skin color and improving skin transparency, which has an effect of improving peripheral blood circulation.

In one embodiment of the present 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 may be selected from the group consisting of softening agents, nutritional lotions, lotions, body lotions, nutritional creams, massage creams, moisturizers, hand creams, essences, eye creams, cleansing creams, cleansing foams, cleansing waters, Gels, patches, lipsticks, makeup bases, and foundations.

The composition of the present invention can have skin aging inhibition and skin wrinkle improving effect, skin whitening effect, skin moisturizing effect and atopy improvement effect. In addition, the composition of the present invention may have an effect of improving skin color and improving skin transparency.

Hereinafter, the present invention will be described in detail.

An embodiment of the present invention provides a composition for external application for skin comprising, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof.

Said "isomer" particularly includes optical isomers, morphoisomers, positional isomers (particularly tautomers) or geometric isomers.

The term "precursor" means that a chemical compound is chemically modified to control physical and chemical properties. Although the compound itself does not exhibit physiological activity, it is converted into an original compound by chemical or enzymatic action in the body after administration. .

The term " pharmaceutically acceptable "as used herein refers to salts which are generally safe, non-toxic, and which are useful for preparing biologically or otherwise preferred pharmaceutical compositions, But also for pharmacological use in humans.

The term "pharmaceutically acceptable salt" as used herein means a salt having pharmaceutically acceptable and desired pharmacological activity as defined above. Such salts include, but are not limited to, (1) salts formed with 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, But are not limited to, benzoic acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, Benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] Which is formed by a carboxylic acid, an ene-1-carboxylic acid, a glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tributyl acetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, Acid addition salts; Or (2) salts formed when acidic protons present in the parent compound are replaced.

The "hydrate" means " pharmaceutically acceptable hydrate ". The "hydrate" is present when the compound of the present invention contains water. The hydrate may contain one or more water molecules per molecule of the compound of the present invention. Exemplary, non-limiting examples include monohydrates, dihydrates, trihydrates, and tetrahydrates. The hydrate may contain one or more compounds of the present invention for one water molecule. Exemplary non-limiting examples include semi-hydrates. In one embodiment, water can be held in the crystal in a variety of ways, such that the water molecule can occupy the lattice position in the crystal or can form a bond with a salt of the compound described above. The hydrate should be "acceptable" in the sense that it is not harmful to its receptacle.

Means a pharmaceutically acceptable solvate, wherein the "solvate" means that the compound of the present invention contains at least one pharmaceutically acceptable solvent. Solvates may contain one or more solvent molecules per molecule of the compound of the present invention or may contain one molecule of the compound of the present invention per molecule of solvent. In one embodiment, the solvent can be maintained in the crystal in a variety of ways, whereby the water molecule can occupy the lattice position within the crystal or can form a bond with a salt of the compound described above.

The above cryptotanshinone can be represented by the following formula (1).

[Chemical Formula 1]

The content of cryptoxanthin may be 0.0001 to 10% by weight based on the total weight of the composition. If the content is less than 0.0001 wt%, the efficacy is insufficient. If the content is more than 10 wt%, the efficacy does not increase remarkably due to an increase in the content, which is not economical.

The cryptoxanthinone may be prepared by organic synthesis or may be obtained from a ginseng extract.

Dansam is a perennial herbaceous plant of the family Lamiaceae (Pinus densiflora, Labiatae), which has tinnitus such as red ginseng, ginseng, and sheep oil. Rhizomes are short and rough, with stalk marks on the ends. The roots are long cylindrical and are divided into several parts. The outer surface is reddish brown or dark reddish brown, rough and has vertical wrinkles. Dansam helps blood circulation, eliminates eosinophilia, and relieves pain in the extremities. Pharmacological actions include increased cardiovascular blood flow, decreased serum lipids, improved microcirculation, antithrombotic action, sedation, antimicrobial action, immune enhancement, anticancer activity, and hypoglycemia. Said ginseng may be, for example, Salvia < RTI ID = 0.0 > miltiorrhiza Bunge ), and the ginseng extract can be obtained, for example, from roots.

The extract as defined in the present invention is characterized by being a crude extract, a polar solvent-soluble extract or a 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 water and ethanol mixed solvent, more preferably 75 to 100% ethanol.

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

More preferably, the extract is firstly extracted with at least one of anhydrous and lower alcohol having 1 to 4 carbon atoms, a polar solvent containing acetone and butylene glycol, and then sequentially extracted with ethyl acetate, diethyl acetate , And a low-polarity solvent including diethyl ether, benzene, chloroform and hexane. More specifically, as an extraction solvent, water, anhydrous and lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, n-propanol, n-butanol), a mixture of water and lower alcohol, acetone, 1,3-butylene glycol and isopropanol And the mixed solvent thereof is subjected to precipitation extraction in an amount of 5 to 20 times by volume based on the dry weight of the ground powder of the root, and the mixture is filtered, and the filtrate is concentrated under reduced pressure. Water is added to the obtained concentrate, and the dispersion is added with one or more of the same amount of low-polarity organic solvent containing ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane and shaken. Thereafter, the organic layer is separated and concentrated under reduced pressure to obtain a root extract.

The extraction may be carried out for about 12 to 96 hours in the case of cold beating, or in an anhydrous or hydrous lower alcohol of 1 to 4 carbon atoms, ethyl acetate, diethyl ether or the like as a solvent at 4 to 25 DEG C for 3 to 20 days The active ingredient can be extracted. In the case of warming, it is preferable that the heating is carried out at a temperature close to the reflux temperature of the solvent for about 5 to 24 hours although it depends on the kind and temperature of the extraction solvent.

By the extraction method which sequentially extracts according to the polarity change as described above, the content of the active ingredient is maximized and the impurity is removed to improve the stability, and the extract having the maximized effect can be obtained.

Filtration is performed after the extraction. In the present invention, filtration is a process of removing suspended solid particles from an extract, and particles, such as cotton, nylon, etc., may be filtered or ultrafiltration, freeze filtration, centrifugation or the like may be used , But is not limited thereto.

In addition, it may further comprise a separation process by various chromatographies (size, charge, hydrophobicity or affinity chromatographic).

Next, the step of drying the filtrate includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying and the like. In some cases, a step of pulverizing the final dried extract may be added.

The composition for external application for skin according to the present invention is generally a concept covering the entire composition used for external 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 into a parenteral dosage form in the form of solid, semi-solid or liquid by adding a compatible inorganic or organic carrier, excipient and diluent with cryptotansinone as an active ingredient. The preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions, and emulsions.

Examples of carriers, excipients and diluents that can be contained in the composition 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 composition for external application for skin according to each formulation, components other than the composition of the present invention may be mixed and selected without difficulty by those skilled in the art depending on the formulation or use purpose of other external preparation for skin, etc. In this case, A synergistic effect can occur.

In addition, when the composition according to the present invention is used as a pharmaceutical composition, it may further contain a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a pharmaceutical adjuvant such as a salt or buffer for controlling osmotic pressure and other therapeutically useful substances And can be formulated into parenteral administration forms according to conventional methods.

It should be understood that the actual dosage of the active ingredient should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration selected, the age, sex, weight and health status of the subject.

Generally, the dosage of active ingredient is in the range of 0.001 mg / kg / day to about 2000 mg / kg / day. A more preferred dosage is 0.5 mg / kg / day to 2.5 mg / kg / day. External administration may be administered once a day or divided into several doses.

In one embodiment of the present invention, the cosmetic composition may be, for example, a basic cosmetic composition, a makeup cosmetic composition, a cosmetic composition for a body and the like, and the formulation is not particularly limited and may be appropriately selected according to the purpose.

For example, the cosmetic composition can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, But is not limited thereto. More specifically, the present invention relates to a cosmetic composition, which comprises at least one of a softening agent, a nutritional lotion, a lotion, a body lotion, a nutritional cream, a massage cream, a moisturizing cream, a hand cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, Cosmetics such as cosmetics such as ointments, ointments, ointments, ointments, lipsticks, makeup bases or foundations, shampoos, rinses, body cleansers, 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 .

When 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. In the case of a spray, in particular, / 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 an enhancer. The thickening agent included in the cosmetic composition of the present invention may be selected from the group consisting of methylcellulose, carboxymethylcellulose, carboxymethylhydroxyguanine, hydroxymethylcellulose, hydroxyethylcellulose, carboxyvinyl polymer, polyquaternium, cetearyl alcohol, Carrageenan, and the like. Among them, at least one of carboxymethyl cellulose, carboxyvinyl polymer, and polyquaternium can be used, and most preferably, it can be a carboxyvinyl polymer.

In one embodiment of the present invention, the cosmetic composition may contain various bases and additives as appropriate, and the kind and amount of these components can be easily selected by the inventors. The composition may contain an additive that is acceptable according to need, and may further include components such as preservatives, pigments, and additives customary in the art.

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

And, in one embodiment of the present invention, the cosmetic composition may comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts. 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.

In addition, the components to be added in addition to the above components are not limited thereto, and any of the above components may be compounded within a range that does not impair the objects and effects of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples, Comparative Examples and Test Examples of the present invention. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

[Example 1] Preparation of Dansamp roots extract

Dansam root was washed with purified water, dried and then hemihydrated. 200 g of the seedling root powder was added to 1 liter of a 70% aqueous ethanol solution, which was boiled for 12 hours in an extractor equipped with a condenser and filtered through a 300-mesh filter cloth. The filtrate was allowed to stand at 4-15 ° C for 7 days and then aged. Then, the filtrate was filtered through Wattman filter paper No. 2. Then, the extract solution was added to a 3 liter separatory funnel, 1 liter of ethyl acetate was added thereto, and the mixture was shaken and mixed thoroughly. After separating completely into two layers, an upper layer (ethyl acetate layer) was taken. The lower layer (water layer) was further extracted twice with a separatory funnel. The separated upper layers were combined, concentrated under reduced pressure to 50 ° C using a distillation apparatus equipped with a cooling condenser, and dried to obtain a dansam root extract having a dry weight of 45.5 g.

[Example 2] Isolation of cryptoxane from dansam root extract

20 g of Dansamp root extract obtained by the method of Example 1 was suspended in a silica gel column (230-400 mesh, ASTM, Art. 9385) and eluted with hexane-methyl chloride (Hexane-CH ₂ Cl ₂ (1: 1) A small fraction was obtained by gradient elution of chloroform (CHCl ₃ ), chloroform-acetone (CHCl ₃ -Me ₂ CO (9: 1)), acetone (Me ₂ CO) and methanol (MeOH) to obtain chloroform- CHCl ₃ -Me ₂ CO (9: 1) eluate was subjected to recycling preperative-HPLC (column, JAIGEL-1H (GPC); flow rate, 3.5 ml / min .; solvent CHCl ₃ , 254 nm) 60.9 mg of yellow cryptotansinone was obtained. Table 1 below shows NMR data for cryptotanse.

Position ^{1 H-NMR (500MHz, CDCl} 3) ¹³ C-NMR (500 MHz, CDCl ₃ ) One 3.22 ( t , 5.1) 29.64 ( t) 2 7.56 ( dd , 7.0, 2.0) 19.03 ( t) 3 7.36 ( d , 7.0) 37.76 ( t) 4 - 34.82 ( s) 5 - 143.67 ( s) 6 8.32 ( d , 9.0) 132.55 ( d ) 7 7.83 ( d , 9.0) 122.48 ( d) 8 X 128.36 ( s) 9 X 126.22 ( s) 10 X 152.35 ( s) 11 - 184.00 ( s) 12 - 175.67 ( s) 13 - 118.26 ( s) 14 - 170.77 ( s) 15 7.31 ( d , 0.5) 81.43 ( d) 16 - 34.56 ( t) 17 2.30 ( d , 1.0) 18.81 ( q) 18 2.70 ( s ) 31.85 ( q) 19 - 31.89 ( q) 20 - -

[Test Example 1] Skin aging inhibition and wrinkle improvement test

1) MMP-1 expression inhibition assay

In the normal human fibroblast monolayer culture system, the cryptoxanthin extract significantly inhibited the expression of MMP-1 induced by UVB 40 mJ / cm ² . Thus, the extract of cryptoxanthin inhibits the biosynthesis of MMP-1, a skin tissue degrading enzyme caused by internal aging or external environmental factors, thereby suppressing skin aging and improving wrinkles. Table 2 below shows the inhibition rate of MMP-1 expression upon treatment with the test substance.

Test substance MMP-1 expression inhibition (%) Control group No treatment 100 Control group UVB 40 mJ / cm ² 152 Cryptoxanthin extract 0.0001 wt% 103 0.001 wt% 71

2) Analysis of Type I Procollagen (Type 1 Procollagen)

In the normal human fibroblast monolayer culture system, the extract of cryptoxanthin showed a distinct stimulatory effect on the production of type Ⅰ procollagen as compared with the control. That is, the extract of cryptoxanthin can inhibit the decrease of collagen production caused by aging of human skin, and exhibits an effect of improving wrinkles. Table 3 below shows the generation of type I < RTI ID = 0.0 > procollagen < / RTI >

Test substance Type I procollagen Generation (%) Control group No treatment 100 Cryptoxanthin extract 0.0001 wt% 110 0.001 wt% 138

[Test Example 2] Skin whitening efficacy test

1) Measurement of inhibition of melanin formation by mouse pigment cells

In order to investigate the inhibitory effect of cryptoxanthin extract on melanogenesis, rat pigment cells were used.

First, the mouse melanoma cells (Dooley, TP et al, Skin pharmacol, 7, pp 188-200) derived from C57BL / 6 mice were inoculated into DMEM (Dulbeccos modified Eagles media) The cells were cultured in a medium supplemented with 100 nM 2-O-tetradecanoyphorbol-13-acetate and 1 nM cholera toxin at 37 ° C and 5% CO 2. The cultured Mel-Ab cells were removed with 0.25% trypsin-EDTA, and the cells were cultured at a concentration of 105 cells / well in a 24-well plate. From the second day, 1 ppm and 10 ppm cryptotans The rice extract was added and cultured. At this time, hydroquinone was used as a control group. After the culture medium was removed, the cells were washed with PBS, and the cells were dissolved with 1 N sodium hydroxide. The absorbance at 400 nm was measured, and the rate of inhibition of melanin production was calculated according to the following equation (1) Method).

Test substance Melanin production inhibition rate (%) Cryptotansinone extract 0.0001 wt% 13.0 Cryptotansinone extract 0.001 wt% 34.0 Hydroquinone (control group) 42.9

As shown in Table 4, the cryptotansinone extract of the present invention showed an excellent inhibition of melanin production and a suppression rate of melanin formation similar to that of hydroquinone. Thus, it was found that the cryptotansinone extract of the present invention had an excellent whitening effect.

2) Test for whitening effect on human skin

The following experiment was conducted to examine the whitening effect of the extract of cryptoxanthinone on the human skin.

At first, 10 healthy men were attached to opaque tape with a hole of 1.5 cm in diameter at the upper abdomen of the subject. Ultraviolet rays (UVB) of about 1.5 to 2 times of the minimum erythema dose of each subject, To induce blackening of the skin.

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

Then, the difference (ΔL *) between the starting time of application of the respective test substances and the completion time of the application of the test materials was calculated according to the following equation, and is shown in Table 5 below. On the other hand, the whitening effect is judged by comparison of DELTA L * between the sample application site and the control site. When the DELTA L * value is 2, the whitening of the deposited pigment is noticeable. .

Test substance The degree of skin color brightness (DELTA L *) Cryptoxanthin extract 1.70 ± 0.20 Hydroquinone (positive control) 1.87 + - 0.11 Vehicle (negative control) 0.40 + 0.14

As shown in Table 5, it was confirmed that the cryptotansinone extract of the present invention showed a degree of skin color brightness similar to that of hydroquinone. This is because the cryptotansinone extract improves pigmentation caused by ultraviolet rays to brighten skin color.

[Test Example 3] Skin moisturizing and atopic symptom improvement test

1) Differentiation induction test of human keratinocyte

The amount of CE (Cornified Envelope) produced during differentiation of keratinocyte and human skin cell line (HaCaT) was measured and the absorbance was used to examine the effect of cryptanthocyanin extract on cell differentiation.

The primary cultured human keratinocytes were placed in a culture flask and adhered to the bottom. The test substances of the following Table 6 were added to the culture medium at the concentrations shown in Table 6, ≪ / RTI > to about < RTI ID = 0.0 > 80% < / RTI > The cells were harvested and washed with PBS (phosphate buffered saline). The cells were washed with 10 mM Tris-HCl buffer (Tris-HCl buffer, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiothreitol) , pH 7.4), sonication, boiling, and centrifugation were suspended again in 1 ml of PBS, and the absorbance at 340 nm was measured. Separately, a portion of the solution after the sonication was taken to measure the protein content and used as a criterion when evaluating the degree of cell differentiation. The test results are shown in Table 6 below, in which the low calcium (0.03 mM) and high calcium (1.2 mM) treated groups were used as negative / positive control groups and the test substance was added to the low calcium concentration.

Test substance The ability to differentiate in keratinocytes (%) Control group Low Ca < ^{2 +} & gt ^; (0.03 mM) 100 Hign Ca < ^{2 +} & gt ^; (1.2 mM) 210 Cryptoxanthin extract 0.0001 wt% 113 0.001 wt% 122 0.01 wt% 128

As shown in Table 6, the cryptotansinone extract according to the present invention promotes differentiation in keratinocytes. Therefore, the extract of cryptoxanthin has skin moisturizing effect.

2) Expression of transglutaminase in human skin cell line

Transglutaminase is a kind of skin differentiation index substance, which helps the formation of CE (Cornified Envelope) and cell differentiation, which are generated during the differentiation of human skin cell line (HaCaT), to be smooth.

Human skin cells to each well in 96 cell culture plate welhyeong 5x10 into ^four was attached for 24 hours. After the adhered skin cell line was treated with the test substance, after 2 days, the medium was removed and stored in a -20 ° C refrigerator. Cells treated with the substance were treated with acetone: ethanol (1: 1, v / v) stored at -20 ° C for 30 minutes at 4 ° C, followed by freezing-thawing twice Fix the cells. (1% bovine serum albumin), transglutaminase antibody, HRP anti-mouse antibody (secondary), and the color development was performed using OPD (o -phennyldiamine). Absorbance was measured at 490 nm and calibration was performed by measuring the background at 630 nm. The test results are shown in Table 7 below, in which low calcium (0.03 mM) treated group and high calcium (1.2 mM) treated group are negative / positive control group, and low calcium concentration test substance is added.

Test substance Expression amount (%) of transglutaminase Control group Low Ca < ^{2 +} & gt ^; (0.03 mM) 100 Hign Ca < ^{2 +} & gt ^; (1.2 mM) 142 Cryptoxanthin extract 0.0001 wt% 139 0.001 wt% 180 0.01 wt% 211

As shown in Table 7, the cryptotansinone extract of the present invention showed an increase of about twice as much as that of the control group, and the amount of the extract was larger than that of the control group. Therefore, it was found that the cryptotansinone extract according to the present invention improves the expression of transglutaminase.

3) Improvement of atopic dermatitis

The effect of improving atopic symptoms was measured as follows. Forty patients between the ages of 10 and 50 who were diagnosed with atopic symptoms (itch, eruption, severe dryness) or who had atopic symptoms by visual observation were divided into two groups, and each of Example 1 and Comparative Example 1 And applied to the area showing atopic symptoms twice a day for 12 weeks.

Table 8 below shows the compositions of Example 1 and Comparative Example 1.

ingredient Example 1 Comparative Example 1 Content (% by weight) Content (% by weight) Cryptoxanthin 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

The efficacy evaluation was conducted after 12 weeks' treatment, and the degree of improvement was judged subjective by the questionnaire. In the questionnaire, symptoms of atopic dermatitis were divided into "itching", "dryness", "keratinization", "dandruff", "erythema", "swelling", "skin crack" In the presence of symptoms, the degree of improvement was assessed for each symptom, and then the average improvement was evaluated for individual atopic symptoms. The results are shown in Table 9 below.

Test group Number of respondents (in) Significant improvement Improving No change worse Cryptotansinone extract (Example 1) 6 8 6 0 Control group
(Comparative Example 1) 2 4 14 0

As shown in Table 9, it can be seen that Example 1, which is a treatment group applied with the formulation containing the cryptotansinone extract, has more general atopic symptoms than Comparative Example 1 which is a control group.

[Test Example 4] Skin color improvement test

1) NO production in HUVEC (Human umbilical vein endothelial cell) of cryptoxanthin extract

Endothelial nitric oxide synthase (eNOS) is present in the vascular endothelial cells of humans, and its activity is increased to generate nitric oxide (NO) to expand blood vessels and promote blood circulation. Human vascular endothelial cells were cultured and the amount of NO produced by treatment with cryptotansinone extract was compared. Table 10 below shows NO production by the cryptoxanthin extract.

Test substance NO (nitric oxide) production ability (%) Control group Low Ca < ^{2 +} & gt ^; (0.03 mM) 100 Cryptoxanthin extract 0.005 wt% 320 0.01 wt% 780 0.02 wt% 1190

As can be seen from the results of Table 10, the cryptotansinone extract showed excellent NO production ability in vascular endothelial cells. The excellent NO (nitric oxide) production ability of the cryptotansinone extract as described above can expand the capillary blood vessels and promote blood circulation, thereby smoothly supplying nutrients to the skin, inhibiting skin aging, and improving the skin color . It also contributes to skin transparency improvement efficacy.

2) Test of improvement of color of human skin (face)

In order to evaluate the effect of the composition containing the cryptotansinone extract of the present invention on the promotion of blood circulation in the skin, the degree of blood circulation in the skin was measured using Laser Doppler Perfusion Imager (LDPI) Is a device that is widely known and currently used as a device for measuring blood circulation. It is a highly sensitive device capable of measuring not only the velocity and amount of blood in the capillary blood vessels of the skin, but also the flow in the small arteries and the range vessels.

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

For use in the test, Example 1 with the composition of Table 8 above was prepared and the blood flow and skin temperature after 1 week of use of Example 1 were compared with initial measurements. Table 11 shows the LDPI results before and after the use of Example 1, and Table 12 shows the IR results before and after use.

Face Before use After 1 week use medium 0.952 1.169

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

As described above, the use of Example 1 containing the extract of cryptotansinone showed an improvement in blood circulation as an effect of improving peripheral blood circulation on the face, which ultimately effectively transmits nutrients of the skin and inhibits and delays skin aging .

Formulation examples of the cosmetic composition and the pharmaceutical composition according to the present invention will be described below. However, the present invention can be applied to various formulations other than the following examples.

[Formulation Example 1] Flexible lotion (skin lotion)

Flexible lotion was prepared by a conventional method according to the composition shown in Table 13 below.

ingredient Content (% by weight) Cryptoxanthin 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)

Nutrition lotion was prepared according to a conventional method according to the composition shown in Table 14 below.

ingredient content
(weight%) Cryptoxanthin 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 / capriciculiglyceride 5.0 Squalane 5.0 Sorbitacecequioleate 1.5 Cetearyl alcohol 1.0 Triethanolamine 0.2 Preservative, fragrance Suitable amount Purified water Balance system 100

[Formulation Example 3] Nourishing cream

Nutrition creams were prepared according to the compositions shown in Table 15 below in a conventional manner.

ingredient content
(weight%) Cryptoxanthin extract 1.0 glycerin 3.5 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capriciculiglyceride 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

Massage creams were prepared according to the compositions shown in Table 16 below by a conventional method.

ingredient Content (% by weight) Cryptoxanthin extract 1.0 glycerin 8.0 Butylene glycol 3.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Wax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 paraffin 1.5 Preservative, coloring, fragrance Suitable amount Purified water Balance system 100

[Formulation Example 5] Pack

Packs were prepared in a conventional manner according to the composition shown in Table 17 below.

ingredient Content (% by weight) Cryptoxanthin 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] Drugs for topical administration of patches

The patches were prepared in a conventional manner according to the compositions shown in Table 18 below.

ingredient Content (% by weight) Cryptoxanthin extract 1.0 Beta-1,3-glucan 3.0 Diethylamine 0.7 Sodium sulphate 0.1 Polyoxyethylene lauryl ether (E.O. = 9) 1.0 Polyhydroxyethylene cetyl stearyl 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 (9)

A composition for external application for skin for skin whitening, which contains, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient and has an effect of inhibiting melanin production. A composition for external application for skin for moisturizing skin, which contains, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient and has an effect of promoting differentiation of keratinocytes. A skin moisturizing agent which contains, as an active ingredient, cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient and has an effect of increasing the expression of transglutaminase of a skin cell line Composition for external application for skin. A method for improving skin color balance and improving skin transparency, which comprises an effective amount of cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient and has an effect of increasing NO production in vascular endothelial cells Composition for external application for skin. A composition for improving skin color and having an effect of improving peripheral blood circulation, which contains cryptotanshinone, an isomer thereof, a precursor thereof, a salt thereof, a hydrate thereof or a solvate thereof as an active ingredient, Composition. 6. The method according to any one of claims 1 to 5,
Wherein the content of the cryptoxanthin is 0.0001 to 10% by weight based on the total weight of the composition. delete delete delete