KR20180121412A - Whitening cosmetic composition and functional food comprising demethoxycurcumin from Curcuma aromatica - Google Patents

Abstract

The present invention relates to a cosmetic composition and beauty functional food containing demethoxycurcumin (DC) isolated from a Curcuma aromatic extract which has melanin synthesis inhibitory activity. According to the present invention, the microphthalmia-associated transcription factor (MITF) is controlled in melanocytes to suppress melanogenesis, and thus the present invention can be usefully utilized for a cosmetic composition and beauty functional food for skin whitening.

Description

A cosmetic composition comprising dimethoxycucumin isolated from a turmeric extract showing a whitening activity and a cosmetic composition comprising a cosmetic composition and functional food comprising demethoxycurcumin from Curcuma aromatica,

The present invention relates to a whitening cosmetic composition comprising demethoxycurcumin separated from a turmeric extract and a cosmetic food.

In cosmetics, skin whitening is one of the areas of greatest interest among East Asian women, especially in Korea, Japan and China. The most important factor in skin whitening is skin melanin pigment.

Originally, melanin is a black pigment found in animals, plants and microorganisms. It is not essential for growth or development, but it enhances survival and competitiveness in certain environments. In particular, melanin in animals is a cause of skin disease and malignant melanoma DHN melanin, GDHB melanin, and catechol melanin have been reported in addition to DOPA melanin, which is related to melanomas and is produced from tyrosine by tyrosinase (Bell, AA and Weeler MH, Ann. Rev. Phytophathol. 24, 411, 1986). And this melanin pigment is a stable pigment which is insoluble in almost all the solvents and it is not easy to handle melanin or chemical treatment. Because of this, the chemistry of melanin has not been clarified, and the progress of research has been slow. In mammals it is clear that almost the same melanogenesis occurs and occurs in the organelle called the melanosome of specifically differentiated cells called melanocytes.

In other words, skin color is determined by the content and distribution of melanin, and is related to the number and distribution of melanosomes released from the melanocytes in the cells and released to the outside of the cells. Hyperpigmentation of the skin can be caused by various factors such as hormonal abnormalities in the body, genetic diseases, and ultraviolet irradiation after inflammatory reaction of the skin. The main factor is due to melanin pigment synthesis abnormality and distribution abnormality. The main function of melanin is to protect the skin from damage by removing oxygen radicals, which means that melanin has an effective countermeasure to protect skin from physical and chemical toxicants.

Prevention of pigmentation in the skin is possible from three perspectives. 1) development of tyrosinase synthesis inhibitor or tyrosinase substrate antagonist to regulate tyrosinase activity, which is the main enzyme of melanin synthesis; 2) Development of a substance that exhibits toxicity to melanocytes in order to lower the function of melanocyte which is an animal's melanin biosynthesis site, 3) development of a dopa reducing substance and prevention of oxidation of dopa, (DOPA chrome tautomerase), which is a second enzyme that catalyzes the conversion of tyrosinase and DOPA chrome to DHICA, or indole-5,6-quinone- And the control of the activity of a third enzyme that catalyzes the conversion to 2-carboxylic acid (indole-5,6-quinone-2-carboxylic acid). Among them, hitherto known inhibitors of tyrosinase include hydroquinone, resorcinol, 4-hydroxyanisole, ascorbic acid and its derivatives, kojic acid Arbutin, glucosamine, oxyreseveratrol, alpha-viniferin, and ferulic acid. However, due to problems of skin safety and formulation stability, arbutin, And kojic acid are used only in a limited amount as an additive to whitening agents.

Until recently, various plants have been studied for the development of natural whitening agents. Among them, it is known that there is a tyrosinase inhibitory activity, such as alfalfa, rhododendron, licorice, green tea, seaweed and gardenia fruit, and some of them are used in products.

Excessive deposition of melanin may be a cause of pathological problems such as skin, but it is recognized as a cosmetic problem such as spots, freckles, dots, and black spots. Therefore, a whitening agent based on natural substances capable of inhibiting melanin formation Is required.

Korean Patent Laid-Open Publication No. 2013-0001027 discloses a cosmetic composition for whitening comprising an extract of Angora oranges and a method for producing the same. Korean Patent Publication No. 1396275 discloses a Gangjin extract having a skin whitening effect.

The present invention aims to provide a whitening cosmetic composition and a food product derived from a natural substance that can effectively inhibit melanin production without toxicity.

In one embodiment, the present invention provides a cosmetic composition or cosmetic for skin whitening characterized by inhibiting the production of melanin containing dimethoxacucumine or a cosmetically acceptable salt thereof as an active ingredient.

The dimethoxycucumine contained in the composition according to the present application is derived from sulfuric acid.

The composition according to the present invention may exhibit a whitening effect by inhibiting melanin biosynthesis through interfering with tyrosinase, particularly including synthesis of tyrosinase enzyme, inhibition of enzyme activity, and / or promotion of degradation.

In another aspect, the present invention also provides a pharmaceutical composition for the treatment of pigmented skin diseases comprising melanin hypercholesterolemia, which comprises inhibiting melanin production comprising dimethoxycucumine or a pharmaceutically acceptable salt thereof as an active ingredient to provide.

Since the composition comprising dimethoxycucumine according to the present invention can inhibit melanin biosynthesis inhibition and expression or activity of various enzymes including tyrosinase involved in biosynthesis of melanin, It is apparent that the present invention can be effectively used for the treatment or prevention of pigmented skin diseases including asthma. Such diseases include, for example, scarring or dermatitis including freckles, senile plaques, dullness, liver, spots, brown or black spots, daylight pigment spots, bluish schizophrenia, hyperpigmentation after drug use, pregnancy brown spots, or scratches and burns , ≪ / RTI > post-inflammatory hyperpigmentation due to < RTI ID = 0.0 >

In another embodiment, the present disclosure also relates to dimethoxycucumine or a pharmaceutically acceptable salt thereof; Or melanin extract of the present invention as an active ingredient. The present invention also provides a health functional food composition for improving skin melanin pigment over-deposition.

In another embodiment, the present application is also directed to dimethoxycucumine or a pharmaceutically acceptable salt thereof; Or a turmeric extract as an active ingredient. The present invention also provides a functional food composition for skin whitening, which inhibits the production of melanin.

In another embodiment, the present application also provides a method of melanin-producing promyelandin hypercholesterolemia in Invitro comprising the step of treating dimethoxacucumine with a cell capable of producing melanin.

Demethoxycurcumin according to the present invention can be effectively used as a cosmetic composition and cosmetic composition having whitening function because it shows excellent whitening efficacy unlike curcumin which is highly cytotoxic and does not exhibit side effects even in a human subject safety test have.

Figure 1 shows the results of cytotoxicity test for melamine of curcumin. Coccumin has been shown to kill 80% of cells at all treated concentrations, especially at 30 μM, indicating that it may be difficult to apply to actual skin for curcumin whitening activity.
Figure 2 shows the structural formula of methoxycucumine according to the present invention.
FIG. 3 shows the results obtained by treating the mouse melanocyte cell line Melan-a by the concentration of dimethoxacucum (5, 10, 20, 30 μM) isolated from a turmeric extract. This indicates that the dimethoxycarcum according to the present invention is not toxic even at a concentration of 30 μM, which shows severe toxicity of curcumin, and can effectively inhibit melanin pigment formation.
Figures 4A and 4B show the inhibition of TRP-1, -2 and tyrosinase protein expression during 24 hours (Figure 4A), 48 hours (Figure 4B) treatment of dimethoxacucum by concentration (5,10,20, This is a photograph showing the effect.
5A, 5B and 5C) of TRP-1 (FIG. 5A), TRP-2 (FIG. 5B) and Tyrosinase (FIG. 5C) at concentrations of dimethoxycucumine (5, 10, 20, 30 μM) Expression inhibition effect.
6A and 6B are graphs showing the effect of inhibiting MITF protein (FIG. 6A) and gene expression (FIG. 6B) at different concentrations of dimethoxacucum (5, 10, 20, 30 μM).
FIG. 7 is a graph showing the inhibitory effect on the p-CREB DNA binding activity by the concentration of dimethoxacucum (5, 10, 20, 30 μM).
FIG. 8 is a graph showing the inhibitory effect of p-AKT and p-ERK on the activity of dimethoxacucum by concentration (5, 10, 20, 30 μM).
FIG. 9 shows the effect of inhibiting melanin pigment formation (FIG. 9A) and cell cytotoxicity (FIG. 9B) by treatment with dimethoxacucum by concentration (10, 20, 30 μM) in a 3D artificial skin model to be. In particular, Figure 9B shows no cytotoxicity compared to curcumin at all concentrations treated compared to Figure 1.

The present invention is based on the finding that dimethoxycarcum separated from turmeric in comparison to curcumin is safe because it is free from cytotoxicity and that it acts as a tyrosinase inhibitor, Based on the discovery that it is possible to solve and sustain a skin whitening effect effectively.

In one embodiment, the present invention is directed to a process for the preparation of demethoxycurcumin 1- (4-Hydroxy-3-methoxyphenyl) -7- (4-hydroxyphenyl) -1,6-heptadiene- ) Or a cosmetically acceptable salt thereof as an active ingredient to inhibit the production of melanin.

In one embodiment according to the present disclosure, Dimethoxycucumine can be extracted from Curcuma aromatica using the method described herein. Dimethoxycucumin according to the present invention can be applied to skin whitening compositions by confirming its action as a tyrosinase activity inhibitor, and demethoxycurcumin having such an effect can be used for skin whitening As a composition, it can be mixed with various cosmetics to provide a whitening effect with a whitening functional cosmetic.

In the present invention, the inhibitory activity of protein, gene expression and tyrosinase, which is a rate-limiting enzyme system of melanin synthesis of demethoxycurcumin, isolated from turmeric extract, and a microphthalmia related mechanism Study.

Tyrosinase (monophenol, dihydroxy-L-phenylalanine: oxygen oxidoreductase, EC 1.14.18.1) is a copper-containing enzyme that inhibits L-tyrosine, L-3,4-dihydroxyphenylalanine- DOPA) (cresolase activity), and DOPA to L-dopaquinone (catecholase activity). The reactions are summarized as follows:

Tyrosine + Dihydroxyphenylalanine + O ₂ →

Dihydroxyphenylalanine + DOPA-quinone + H ₂ O

The dimethoxycucumine of the present invention is a melanocyte that contains the tyrosinase and tyrosinase-related protein-1 and -2 (TRP-1 , -2) and protein expression and gene expression. In addition, the microphthalmia-associated transcription factor (MITF) is a transcription factor that plays an important role in melanogenesis and diminishes the expression of proteins and genes by dimethoxacucumine. It was also confirmed that inhibition of protein expression of SRY-related HMG-box 10 (SOX10), an important transcription factor of MITF, and inhibition of gene expression of MITF by inhibiting the DNA binding ability of phospho-cAMP response element binding protein (p-CREB) Respectively. The expression of p-AKT and p-ERK in mitogen-activated protein kinase (MAPK) signaling, which affects the expression of MITF, was also observed to suppress protein expression of p-AKT and p-ERK when treated with DC It can serve as a candidate for inhibition of whitening and pigmentation, which is useful as a whitening candidate material and also exhibits melanin synthesis inhibitory effect in a 3D artificial skin model.

As used herein, the term " skin whitening " is intended to inhibit changes in skin color or pigmentation caused by excessive production of pigment in the skin due to external environmental changes such as ultraviolet rays, stress, hormone secretion abnormality and pregnancy. The effect of improving or preventing or preventing dark circles, spots, freckles and dark circles occurring, the effect of increasing the transparency of the skin, the effect of improving the dullness and tightness of the skin by improving the dullness of the skin.

In general, pigmentation of the skin is stimulated by ultraviolet light or hormone imbalance, leading to melanocyte stimulation, which is the main cause of skin deposition of melanin pigment biosynthesized. Therefore, compositions according to the present invention capable of inhibiting melanin biosynthesis in melanocyte, which is a melanin biosynthesis cell, can be used in various applications requiring inhibition of pigment deposition other than the above-mentioned whitening.

The composition according to the present disclosure may also comprise a cosmetically acceptable salt of dimethoxycucumine. This is useful for acid addition salts formed by free acids. Acid addition salts can be prepared in a conventional manner, for example, by dissolving the compound in an excess amount of an aqueous acid solution and precipitating the salt with a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration. As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acids include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, trifluoroacetic acid, The organic acid may be selected from maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycollic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid and hydroiodic acid can be used. It is not limited.

As used herein, the term cosmetic composition refers to an article used in the human body to brighten or darken the appearance of a human body to improve appearance or to maintain or promote the health of skin and hair, and refers to a slight action on the human body. Commonly used include lotions, creams, oils, cleansing products, and functional cosmetics. Functional cosmetics are physical control, biochemical, and biotechnological techniques that control the body's defense rhythm, cosmetic control, disease control and recovery, And cosmetics which are processed and designed so that the functions are sufficiently expressed in living bodies.

The cosmetic composition or cosmetic according to the present invention may be provided, for example, as a cream, a lotion, an ampoule, a skin, an essence, a soap, (Body lotion, lipstick, mascara, make-up base), or in the form of a mask or pack.

The site to which the cosmetic composition or cosmetic according to the present invention can be applied is not limited to the specific fat tissue of the body, but can be applied to various parts of the skin, especially face, arm, leg, etc., which require inhibition of pigmentation or whitening. The site of application may be administered by a variety of routes of administration, such as systemic or topical, especially topical, transdermal, or injection, particularly the transdermal route.

The cosmetic composition according to the present invention has no particular limitation on the other components other than the dimethoxycucumine of the present invention as essential components in the indicated ratios and can be used in the case of ordinary cosmetics such as cream, vegetable oils, emulsifiers, thickeners, Water, an antioxidant, and a UV absorber.

The content of dimethoxycucumine in the composition according to the present invention may vary depending on the specific use or formulation of the desired product, but may be about 0.001 to about 50% by weight in the total composition, It does not.

The cosmetic composition according to the present invention may contain conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments and / or fragrances in addition to dimethoxycinnamic acid.

The cosmetic composition of the present invention may comprise a water-soluble base material comprising an unsaturated fatty acid or propylene glycol including conjugated linoleic acid, linoleic acid, gamma-linoleic acid, alpha-linoleic acid.

In addition, the cosmetic composition of the present invention may be prepared in any form conventionally produced in the art, and may be prepared, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a mask, a pack, a spray or a powder.

When the formulation of the present cosmetic composition is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .

When the formulation of the present cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, a mixture of chlorofluorohydrocarbons, Propane / butane or dimethyl ether.

The cosmetic composition according to the present invention may further comprise a cosmetically / dermatologically acceptable carrier. &Quot; cosmetically / dermatologically acceptable " means suitable for use in contact with a tissue (e.g., skin or hair) without undue toxicity, insolubility, instability, irritation, allergic response,

When the formulation of the composition is a solution or an emulsion, a solvent, a solubilizing 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, propylene glycol, , 3-butyl glycol, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan can be used.

When the formulation of the present composition is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension 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 composition of the present invention is an interface-active agent-containing cleansing, the carrier component is selected from the group consisting of aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolium derivatives, methyltaurates, sarcosinates, fatty acid amide ethers Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester. According to a preferred mode of administration, the compositions of the present invention may comprise at least one pharmaceutically acceptable excipient, especially a dermatologically acceptable excipient.

Further, in the composition for dermatological preparation of each of the formulations, components other than the above-mentioned essential components may be selected and mixed without difficulty by those skilled in the art depending on other compositions for dermal use or the purpose of use.

Since the composition of the present invention has little toxicity and side effects, it can be safely used for prolonged use for preventive purposes.

As described above, the dimethoxycacinum according to the present invention can essentially suppress the melanin biosynthesis and can be used for the treatment of diseases associated with various hypermolanin pigment deposition requiring inhibition of melanin biosynthesis.

As used herein, the term " hyperpigmentation ", including melanotic hypercholinism, refers to darkening or darkening of the skin or other areas of the hand or toenails due to excessive increase of melanin at certain sites. The melanin pigment hyperpigmentation disorder may be selected from the group consisting of freckles, dullness, senile spots, liver, spots, brown or black spots, daylight pigmentation, cyanicmelasma, hyperpigmentation after drug use, gravidic chloasma, And post-inflammatory hyperpigmentation due to burns or dermatitis, including burns, and the like.

Pharmaceutically acceptable salts herein refer to those which are physiologically acceptable and do not cause an allergic reaction or similar reaction that is common in humans, and the salt is preferably an acid addition salt formed by free acid Do. The free acid may be an organic acid or an inorganic acid. The organic acids include, but are not limited to, citric, acetic, lactic, tartaric, maleic, fumaric, formic, propionic, oxalic, trifluroacetic, benzoic, gluconic, methosulfonic, glycolic, succinic, Glutaric acid and aspartic acid. In addition, the inorganic acid includes, but is not limited to, hydrochloric acid, bromic acid, sulfuric acid, and phosphoric acid. In one embodiment according to the present disclosure, the pharmaceutically acceptable salt may be present as an acid addition salt wherein the dimethoxycucumine compound according to the present invention forms a salt with a free acid. Further, the dimethoxycucumine according to the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates which can be prepared by conventional methods.

As used herein, the term " treatment " refers to any action that improves or alleviates melanotic hypercholesterolemia with administration or application of the composition. Those skilled in the art will be able to ascertain, by reference to the data provided by the Korean Medical Association, the precise criteria of the disease for which the composition of the present invention is effective, .

The pharmaceutical compositions comprising dimethoxycucumine according to the present invention may be formulated with suitable excipients such as suitable carriers, diluents, preservatives, stabilizers, wetting agents, emulsifiers, solubilizers, sweeteners, colorants, As shown in FIG. Specific examples include lactose, dextrose, sucrose, sorbitol, mannitol xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone Water, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium, stearate, mineral oil and the like.

The method of administration of the pharmaceutical composition comprising dimethoxycucumine according to the present invention can be easily selected according to the formulation and can be administered to a human in various routes. For example, it may be formulated in the form of powders, tablets, pills, granules, dragees, hard or soft capsules, liquids, emulsions, suspensions, syrups, elixirs, external preparations, suppositories, sterilized injection solutions, Orally or parenterally. In particular, oral administration is preferred.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid formulations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used diluents, various excipients such as wetting agents, sweeteners, have.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

Further, the pharmaceutical compositions comprising the extract according to the present invention may be prepared using the appropriate methods known in the art or by methods disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA) And the like.

The dosage of the pharmaceutical composition comprising the extract according to the present invention or the active ingredient compound isolated therefrom may vary depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, The effective dose of the extract is usually about 1 to 100 g / day, especially about 10 to 50 g / day, more preferably about 30 g / day for an adult (60 kg). It will be apparent to those skilled in the art that doses may be additive or subtracted, as the dosage can vary depending on various conditions, and thus the dose is not intended to limit the scope of the invention in any way.

The number of administrations can be administered once or several times a day within a desired range, and the administration period is not particularly limited. In addition, the extract of the present invention or a composition comprising the active ingredient compound isolated therefrom may be added to any food in addition to oral administration as it is, and may be routinely ingested.

The pharmaceutical composition comprising dimethoxycucumine according to the present invention not only provides excellent therapeutic effect on pigment deposition through inhibition of melanin biosynthesis but also can be used safely even when taken for a long time without toxicity and side effects caused by drugs.

Accordingly, the dimethoxycucumine according to the present invention can be used as a main ingredient, a minor ingredient, a food additive for food, or a health functional food material such as a whitening functional food.

As used herein, the term " food " means a natural or processed product containing one or more nutrients, preferably a state of being ready to be eaten through a certain degree of processing, It is intended to include food, food additives, functional foods and beverages as a meaning.

As used herein, the term " functional food " refers to a food group that has been imparted with added value to function or express the function of the food by physical, biochemical, or biotechnological techniques, , And the body control function of disease prevention and recovery is designed to be fully expressed in the living body, and in particular, includes "health functional food".

As used herein, the term " health supplement food " is a food prepared by using raw materials or ingredients having useful functions in the human body, and is useful for health and other purposes such as controlling nutrients and physiological actions on the structure and function of the human body For the purpose of health assisting to obtain the effect, it refers to a food which is made by processing a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. ingredients contained in a food raw material.

The functional food and the health supplement food may further include a food-acceptable food supplementary additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of a functional food.

Foods to which dimethoxycucumine according to the present invention can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and functional foods. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods But are not limited to, fruits, vegetables, beverages, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.). The food, beverage or food additive may be prepared by a conventional production method.

In one embodiment, the functional beverage is made of a functional beverage. The functional beverage includes a beverage group to which the added value is imparted so as to function and express the function of the beverage to a specific purpose, physical or biochemical, Defensive rhythm control, disease prevention and recovery, etc., means a beverage which is designed to sufficiently express the body control function against the living body, and particularly refers to a drink for improving skin elasticity and promoting collagen synthesis.

The food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate, etc.), pectic acid and its salts, Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination.

It is to be understood that both the foregoing general description and the following detailed description of the present invention are exemplary and explanatory and are intended to be illustrative of the invention and are not intended to limit the scope of the claims. It will be apparent to those skilled in the art that such variations and modifications are within the scope of the appended claims.

Example

Example  One. Of demethoxycurcumin  detach

1-1. curcuma  Preparation of extract

20kg of dried Uchum (herb medicine circulation) was cooled in MeOH 4 times for 3 days, and the extract was concentrated under reduced pressure to obtain 2.3Kg of MeOH extract.

1-2. curcuma  Isolation of the compound from the extract

750 g of the above MeOH extract 1-1 was partitioned with CHCl ₃ and distilled water, and the CHCl ₃ layer was concentrated under reduced pressure to obtain 500 g of CHCl ₃ soluble fraction. Six fraction fractions were obtained by silica gel column chromatography using hexane / EtOAc mixed solvent as eluent and 200g of CHCl ₃ soluble fraction by stepwise gradient elution. Among them, the fourth fraction having the highest melanin biosynthesis inhibitory activity was further divided into three fractions to obtain a second fraction having whitening activity, and demethoxycurcumin (9 g) showing the property values as described below was isolated The structure was confirmed by comparing it with the physical properties described in the existing literature.

Demethoxycurcumin (Figure 2)

1- (4-Hydroxy-3-methoxyphenyl) -7- (4-hydroxyphenyl) -1,6-heptadien-3,5-

: Yellow powder;

mp 168-169 [deg.] C ESI-MS m / z: 337.1 (M-H)

(2H, d, J = 16.0 Hz, H-3, 3 '), 7.57 (2H, d, J = 8.8 Hz, H-7,9) (1H, d, J = 8.0 Hz), 7.33 (1H, s, H-6 '), 7.16 ), 6.77 (1H, d, J = 16.0 Hz, H-4), 6.70 (1H, d, J = 16.0 Hz, H- s, OMe-7 ');

(S, C-2), 183.1 (s, C-2), 159.8 (s, C-8 '), 149.4 , 148.0 (d, C-7), 140.7 (d, C-4), 140.4 (d, C-4 '), 130.3 , 125.8 (s, C-5 '), 123.1 (d, C-10), 121.0 (d, C-9), 111.3 (d, C-6), 100.9 (d, C-1), 55.7 (OMe, C-7 ').

Example  2: Melanin formation inhibitory effect assay and cytotoxicity test

The melanin production inhibitory effect of the sample obtained in Example 1 was analyzed as follows.

2-1. Animal cell line  culture

The Melan-a cell line, which is a melanocyte derived from an animal cell line used in the present invention, was purchased from Dr. DC Bennett (University of London, UK), and 2 mM glutamine, 100 (100 μg / ml streptomycin, Gibco, USA), 10% fetal bovine serum (Gibco, USA), 100 nM TPA (12-O-tetradecanoylphorbol (Gibco, USA) medium containing 10% carbon dioxide and -13-acetate, Sigma, USA) was used for the experiment while subculturing in a CO2 incubator (CO2 incubator) at 37 ° C in 10% carbon dioxide and 90% air.

2-2. Cytotoxicity test

Cytotoxicity assays were performed by MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay. 0.5 mg / ml MTT was added. After 4 hours, the supernatant was removed, and the formazan produced by 100% DMSO was dissolved. The absorbance at 570 nm was measured and the cell viability (%) was calculated in comparison with the control.

2.3. Analysis of whitening activity by in vitro test

In order to confirm the activity of the sample obtained in the above examples in vitro, experiments were conducted as described below by applying the method described in the literature.

2-2-1. Melanin synthesis inhibitory efficacy

It is a test method that compares the amount of melanin production in the cell with the amount of the blank in cell culture.

Demethoxycurcumin (DC) derived from turmeric was added to the culture medium of melan-a cells, which is the melanocyte of mouse, to test the effect of improving whitening at the cell level.

More specifically, Melan-a cells, which are melanocytes of mice, were divided into 6-well plates at a concentration of 3 × 10 ⁴ cells / well and cultured for 48 hours, And then treated with dimethoxacucum to final concentrations of 5, 10, 20, 30 μM and cultured for 48 hours. Cultured cells were washed twice with phosphate buffered saline (DPBS) containing no calcium chloride (CaCl ₂ ) or magnesium chloride (MgCl ₂ ) and treated with trypsin (Gibco, USA) The same number of cells per group were collected. The cell precipitate was washed with phosphate buffered saline (DPBS) containing no calcium chloride (CaCl ₂ ) or magnesium chloride (MgCl ₂ ), and 100 μl of 1 normal concentration added with 10% dimethylsulfoxide (DMSO) (N) solution of sodium hydroxide (NaOH), treated at 60 DEG C for 30 minutes, absorbance was measured at 475 nm, and the amount of melanin was calculated from a standard curve prepared with synthetic melanin.

As shown in FIG. 3, the concentration of dimethoxacucum was 35.3% at 10 μM, 56.7% at 20 μM and 66.1% at 30 μM, respectively, as compared with the control, (IC50, 17.1 [mu] M).

2-2-2. Melanosite  In a cell Melano Genesis  Evaluation of related protein and gene expression inhibition efficacy

Inhibition of tyrosinase and tyrosinase related protein (TRP) -1, -2 protein and gene expression in melanocyte melanocytes of mice. For efficacy evaluation, DCs were treated with Melanie-A cells for 24 hours and 48 hours, respectively. Similar to the results of melanin content analysis, the expression of tyrosinase, TRP-1, and -2 in both DCs was markedly decreased (FIG. 4A, FIG. 4B). In addition, as a result of the treatment of melan-a cells with DC for 24 hours, concentration of tyrosinase, TRP-1, and TRP-2 decreased in a concentration-dependent manner (FIGS. 5A, 5B and 5C).

2-2-3. Of DC Microphthalmia -associated transcription factor ( MITF ) Protein and Gene Expression Suppression Efficacy

As a result of treatment of DC with Melan-a cells by concentration, protein expression of MITF, which is a factor controlling the expression of TRP-1, -2 and tyrosinase, decreased in a concentration-dependent manner (Fig. 6A) Concentration-dependent reduction of MITF gene expression at 20.3% at 20 [mu] M, 23.7% at 20 [mu] M and 83.1% at 30 [mu] M (Fig. 6B).

2-2-4. The p- CREB  binding inhibition efficacy

Melan-a cells were treated with DC for 24 hours, and protein expression and DNA-binding activity of p-CREB, a major transcriptional leader of MITF, were observed. As a result, the expression of p-CREB DNA was not changed, but the p-CREB DNA binding activity was decreased in concentration-dependent manner by 22.7% at 10 μM, 34.8% at 20 μM and 47.0% at 30 μM (Fig. 7).

2-2-5. p- ERK , p- AKT  Protein activity inhibitory effect

The expression of p-AKT and p-ERK in mitogen-activated protein kinase (MAPK) signaling, which affects the expression of MITF, , and protein expression of p-AKT and p-ERK decreased in a concentration-dependent manner (Fig. 8).

2-3. Using a 3D artificial skin model DC processing  Assessment of postmelanine synthesis inhibitory efficacy and cytotoxicity

3D artificial skin model, including melanocytes (Neoderm ^®, Tego Science, by the DC of the Republic of Korea treatment 4 days by concentration results that the concentration of DC 14.2% at 10μM, 20.1% at 20μM and became a melanin synthesis inhibition by 24.4% at 30μM (FIG. 9 (A)). The cytotoxicity experiment using MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) Indicating that the inhibition of melanin synthesis by the cells was not due to toxicity and also showed that the cells were not cytotoxic, indicating that DC is not cytotoxic compared to curcumin at all concentrations specifically treated as compared to FIG.

Example  3: Human skin primary stimulation test

3-1. Purpose of examination

This test was conducted to confirm the primary stimulation of dimethoxicacumin on human skin.

3-2. Test substance

3-3. Test Methods

3-3-1. Subject

A total of 30 participants participated in the entire study. The average age of the subjects was 40.70 ± 8.0, with the highest age at 50 and the lowest age at 22. The skin characteristics of the subjects were investigated by questionnaire (see Table 1-1, Table 1-2).

[Table 1-1] Subjects' skin characteristics (n = 30)

[Table 1-2] Subjects' skin characteristics (continued from Table 3-1)

* Subjects who responded to metal and other (dust) allergies were not expected to have any effect on the test results.

3-1-2. Selection Criteria

Candidates who meet the following conditions are selected as the test subjects.

① Healthy person without any skin disease in men and women aged 18 ~ 60

② Applicants who voluntarily signed a written agreement before he / she has fully explained the purpose and contents of the examination prior to the examination

③ Those who are willing to cooperate well with the requirements of the experiment during the test period and to contact them promptly if there are strange symptoms

④ Applicants who can be followed up during the examination period

3-3-3. Exclusion criteria

Applicants who fall under the following items are excluded from the test subject group.

① Pregnant, breastfeeding, or possibly pregnant women

② If there are tattoos, scars, burns, etc. on the test site and it interferes with scoring.

③ In case of infectious skin disease or other skin disorders that interfere with the purpose of the test

④ If the drug currently being treated affects the skin reaction

⑤ Cosmetics, medicines, or those who are irritated or allergic to daily light exposure

⑥ Those with atopic skin

⑦ Those taking contraceptives, antihistamines, anti-inflammatory drugs

⑧ If you have pouch keratosis or skin graft donation

⑨ Those who have severe irritation or allergies on the adhesive tape

⑩ If you are participating in another test at the same time

⑪ If you have participated in the same previous examination and have not passed more than 4 weeks

⑫ Others In addition to the above items, if it is judged that the clinical trial is difficult due to the decision of the clinical trial supervisor

3-3-4. Limitations

① The subject was not allowed to touch the test area (etc.) while applying the patch.

② When taking or using medicines for the treatment of the body, the examiner was notified.

3-3-5. Number of subjects and calculation basis

The number of subjects was selected from more than 30 persons based on the toxicity test 7 (1) human skin patch test method [Attachment 1] of functional cosmetics screening regulations.

3-3-6. Test suspension and dropout criteria

Subjects who did not comply with the schedule during the course of the study were asked to continue their participation and decided to discontinue the study and to stop the study if there were other side effects,

① If the subject voluntarily withdraws the clinical trial agreement due to sudden accidents, disease or pregnancy.

 ② When the adverse reaction due to the test substance is serious during the test

 ③ If you do not follow the protocol compliance

 ④ Failure of follow-up observation, etc. If it is thought that there is interference with the test execution by the judgment of the other test person

3-4. Test materials and methods

The test materials are as follows: Finn Chambers: SmartPractice, Denmark, Micropore tape: 3M / Medical-Surgical Division, USA, Microman (M250): Gilson, France, Skin Marker: Chemotechnique Diagnostics AB, Sweden

The test method is as follows:

The test area was washed with 70% ethanol and then dried. The test material was applied as provided by the customer. 16 μl of the test substance was dropped into a Finn chamber, placed on the back of the test site, and fixed with a micropore tape. After removing the patches, the test area was marked with a skin marker, and each test site was observed after 30 minutes and 24 hours.

3-5. Criteria

Observations were made 30 minutes and 24 hours after removal of the patch, and skin reactions were evaluated according to the following criteria, which reflected the Frosch & Kligman method (Table 1).

[Table 2] Clinical standard photographs used for visual evaluation of human papillary test

3-6. How to calculate results

48 hours and 72 hours were calculated using the following equation and the results were determined according to the criteria of Table 2 for the average degree of reactivity for each substance.

[Equation 1]

[Table 3] Primary stimulation index for human skin of cosmetics

3-7. result

No skin reactions were observed in the test material during the test period (Table 4).

[Table 4] Human primary stimulation test

3-8. conclusion

This test substance is regarded as a substance of low stimulus category in terms of primary stimulus of human skin.

Hereinafter, a cream composition, a massage cream composition, a lotion composition, a skin lotion composition, an essence composition, a pack composition and a cleansing foam composition are exemplified as the formulations of the present invention. However, the formulation including the cosmetic composition of the present invention is not limited thereto no. In addition, the extract of turmeric as an active substance represents demethoxycurcumin.

Formulation Example 1. Cream composition

The oil phase and water phase were each heated to 75 ° C with the composition and content (% by weight) shown in Table 5 below, and then cooled to room temperature.

[Table 5]

Formulation Example  2. Cream composition

The oil phase and water phase were heated to dissolve and mixed at 75 DEG C with the composition and content (weight%) shown in Table 6 below, and then cooled to room temperature.

[Table 6]

Formulation Example 3. Lotion composition

The oil phase and water phase were heated and mixed at 75 ° C with the composition and content (weight%) shown in Table 7 below, and then cooled to room temperature.

[Table 7]

Formulation Example 4. Skin lotion composition

The water phase and the ethanol phase are prepared and mixed respectively with the composition and the content (% by weight) shown in Table 8 below, and then filtered.

[Table 8]

Formulation Example 5. Essence composition

The water phase and the ethanol phase were respectively prepared by mixing in the composition and the content (% by weight) shown in Table 9, and then filtered.

[Table 9]

Formulation Example 6. Pack composition

The water phase and the ethanol phase are dispersively dissolved and mixed in the composition and the content (% by weight) shown in Table 10 below, and then cooled to room temperature.

[Table 10]

Formulation Example 7 Cleansing Foam Composition

The water phase and the oil phase were dispersed and dissolved by the composition and the content (% by weight) shown in Table 11 below, mixed and sieved, and then cooled to room temperature.

[Table 11]

In addition, examples of formulations for beauty food of the present invention include chewing gum, candy, biscuit, ice cream, sausage, chocolate, ordinary beverage, and whitening probiotic lactic acid bacteria beverage formulation. However, the formulation including the cosmetic food composition of the present invention is not limited thereto no.

Example 1: Preparation of chewing gum (A)

Gum base 20 wt%

Sugar 76.5 wt%

0.5% by weight dimethoxycucumine

Fruit flavor 1 wt%

Water 2 wt%

Chewing gum was prepared in a conventional manner with the above composition and content.

Example 2: Preparation of chewing gum (B)

Gum base 25 wt%

Sorbitol 72 wt%

0.5% by weight dimethoxycucumine

Peppermint Lever 2.5 wt%

Chewing gum was prepared in a conventional manner with the above composition and content.

Example 3: Preparation of candy (A)

54.5 wt%

45% by weight of starch syrup

0.4% by weight dimethoxycucumine

0.1% by weight of orange flavor

Candies were prepared by a conventional method with the above composition and content.

Example 4: Preparation of candy (B)

Maltitol 53.6 wt%

Reduced starch syrup 45%

0.5% by weight dimethoxycucumine

Natural herb fragrance 0.14 wt%

Candies were prepared by a conventional method with the above composition and content.

Example 5: Preparation of biscuit

First class Flour 88 kg

Grade 1 flour 76.4 kg

16.5 kg

Salt 2.5 kg

Glucose 2.7 kg

Palm Shortening 40.5 kg

Ammonium 5.3 kg

0.6 kg of soy sauce

0.55 kg of sodium bisulfite

Rice flour 5.0 kg

Vitamin B1 0.003 kg

Vitamin B2 0.003 kg

Milk flavor 0.16 kg

Water 71.1 kg

Whole milk powder 4 kg

1 kg of replacement milk powder

Calcium Phosphate 0.1 kg

Spray salt 1 kg

Spray oil 25 kg

Dimethoxycucumine 1.7 kg

Biscuits were prepared by the conventional method with the above composition and content.

Example 6: Production of ice cream

10% by weight milk fat

Non-oil solid content 10.8 wt%

Sugar 12.0 wt%

3.0% by weight of starch syrup

0.5% by weight of an emulsion stabilizer (span, span)

Perfume (Strawberry) 0.15 wt%

Water 63.05 wt%

0.5% by weight dimethoxycucumine

Ice cream was prepared in a conventional manner with the above composition and content.

Example 7: Preparation of sausage

Pork 63.6%

Chicken meat 27.5%

Starch 3.5%

Soy protein 1.7%

Salt 1.62%

Glucose 0.5%

Other additives (glycerin) 1.34-1.46%

Dimethoxycucumine 0.12-0.24%

The sausage having the whitening effect was prepared by mixing as described above

Example 8: Preparation of chocolate

Sugar 34.5 wt%

Cocoa Butter 34 wt%

Cocoa mass 15 wt%

Cocoa powder 15 wt%

Lecithin 0.5 wt%

Vanilla flavor 0.5%

0.5% by weight dimethoxycucumine

Chocolate was prepared in a conventional manner with the above composition and content.

Example 9. Preparation of a health whitening probiotic Lactic acid bacteria drink

5% by weight dimethoxycucumine

Probiotics 19.885 wt%

12% by weight isomalt

Powdered crystalline glucose 12 wt%

3% by weight of chicory extract powder

10% by weight of mixed skim milk powder

10% by weight vegetable cream

Xylitol 9 wt%

9% by weight of fructooligosaccharide

Fish collagen 5 wt%

Milk flavor powder 1 wt%

Yoghurt flavor powder 1 wt%

1% by weight of silicon dioxide

1% by weight of magnesium stearate

0.5% by weight of vitamin C

Dry egg yolk powder 0.5 wt%

Enzymatically treated Stevia 0.09 wt%

0.02% by weight of vitamin B2

0.005 wt% folic acid

Example 10. Preparation of whitening beverage

1000 mg of demethoxycurcumin

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (8)

A cosmetic composition for skin whitening characterized by inhibiting the production of melanin comprising dimethoxycucumine or a cosmetically acceptable salt thereof as an active ingredient.
The cosmetic composition for skin whitening according to claim 1, wherein the dimethoxycacin is separated from turmeric.
The cosmetic composition for skin whitening according to claim 1, wherein the inhibition of melanogenesis is by at least one of inhibition of tyrosinase expression, inhibition of tyrosinase activity, or degradation of tyrosinase.
A pharmaceutical composition for the treatment of pigmented skin diseases comprising melanin hypercholesterolemia, which comprises inhibiting melanin production comprising dimethoxycucumine or a pharmaceutically acceptable salt thereof as an active ingredient.
5. The method of claim 4,
The pigmented skin disease may include scarring or dermatitis including freckles, senile spots, dullness, liver, spots, brown or black spots, daylight pigment spots, bluish schistosomiasis, hyperpigmentation after drug use, Wherein the hyperpigmentation is hyperpigmentation after inflammation due to inflammation.
A health functional food composition for improving melanin pigment over-deposition in skin characterized by inhibiting melanin production comprising dimethoxycucumine or a pharmaceutically acceptable salt thereof as an active ingredient.
A functional food composition for skin whitening characterized by inhibiting melanin production comprising dimethoxycucumine or a pharmaceutically acceptable salt thereof as an active ingredient.
A method for inhibiting melanin production in Invitro comprising the step of treating dimethoxacucumine in melanin cells in Invitro.

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