KR101988988B1 - Composition for skin cell regeneration, anti-wrinkle, antioxidant and skin whitening - Google Patents

Abstract

The present invention relates to a composition for regenerating skin, improving wrinkles, antioxidation and skin whitening. Eupatilin according to the present invention promotes collagen synthesis of fibroblasts of the skin to inhibit skin regeneration effect, wrinkle improvement effect, free radical generation, inhibit antioxidative effect and melanin synthesis, , An external preparation for skin, a cosmetic composition or a health food.

Description

TECHNICAL FIELD The present invention relates to a composition for skin regeneration, anti-wrinkle, antioxidant and skin whitening,

The present invention relates to a composition for regenerating skin, improving wrinkles, antioxidation and skin whitening.

Collagen is a major substrate protein produced in fibroblasts of the skin and exists in extracellular epilepsy. Its important functions are mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, division of cells and differentiation Growth or wound healing) are known. Such collagen is reduced by aging and photo aging caused by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin. Also, in recent years, extensive research on skin aging has developed, and important functions of collagen in skin have been revealed.

Effective ingredients promoting collagen synthesis and exhibiting wrinkle-reducing effects are known. For example, retinoic acid, transforming growth factor (TGF) [non-patent document 1], animal placenta-derived protein [Patent document 1], betulinic acid [Patent document 2], chlorella extract [ Patent Documents 3 and 4] are known as collagen synthesis promoting substances. However, the above-mentioned effective ingredients are limited in the use amount due to safety problems such as irritation and redness when applied to the skin, or have insufficient effect, so that the effect of improving the skin function by promoting the collagen synthesis of the skin can not be expected.

On the other hand, active oxygen introduced from the outside of the living body or generated in the living body causes many problems such as promoting aging of the living body, cancer, and the like. Therefore, the development and research of antioxidants that inhibit oxidation by active oxygen have been performed. Antioxidants are widely distributed in copper and plants. Many phenolic compounds, flavonoids, tocopherols, vitamin C and selenium are known in fruits and vegetables. However, the antioxidant substances present in nature can not be expected to have practically sufficient effects in skin application. Therefore, although synthetic antioxidants having excellent antioxidant ability and low cost are widely used, their use is restricted due to safety concerns such as human side effects.

In addition, it is the desire of everyone to have a whiter skin. It is genetically determined by the concentration and distribution of melanin in human skin, but is also influenced by environmental or physiological conditions such as sunlight, fatigue, and stress. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine tyrosine, which is an amino acid, is converted to DOPA or dopaquinone by an enzyme called tyrosinase. Thus, the pathway through which melanin is made is known, but the mechanism by which melanin synthesis, the previous step in which tyrosinase acts, is not yet elucidated.

On the other hand, commonly known whitening ingredients include substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L-Ascorbic acid) There are plant extracts. By inhibiting the synthesis of melanin pigment, they can brighten the skin tone to realize skin whitening, and it is possible to improve skin hypercholesterolemia such as stain or freckles due to ultraviolet rays, hormones or heredity. However, when applied to skin, there is a problem that the use amount is limited due to safety problems such as irritation and redness, or the effect is insignificant, so that a practical effect can not be expected.

Therefore, there is a desperate need for development of ingredients having skin regeneration, wrinkle improvement, antioxidative and whitening activity, which are more safe than the living body, have a stable active ingredient, and are more effective than the substances having existing skin regeneration, wrinkle improvement, antioxidant and whitening effect Is desired.

[Prior Art]

[Patent Literature]

Japan Patent No. 8-231370

Japan Patent No. 8-208424

Japan Patent No. 9-40523

Japan Patent No. 10-36283

[Non-Patent Document]

Cardinale G. et al, Adv. Enzymol., 41, p. 425, 1974

Accordingly, the present inventors have found that Eupatilin promotes collagen synthesis of fibroblasts of the skin and inhibits collagenase activity, thereby promoting skin regeneration, improving wrinkles, eliminating free radicals, And inhibiting the synthesis to show a whitening effect, thus completing the present invention.

Accordingly, an object of the present invention is to provide a composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient:

[Chemical Formula 1]

As means for solving the above problems,

There is provided a pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

 [Chemical Formula 1]

As another means for solving the above problems,

A skin external preparation for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

As another means for solving the above problems,

There is provided a cosmetic composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

As another means for solving the above problems,

There is provided a health food for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

Eupatilin according to the present invention promotes collagen synthesis of fibroblasts of the skin and inhibits collagenase activity thereby promoting regeneration of the skin, eliminating free radicals to exhibit antioxidative effects, inhibiting melanin synthesis, It can be used for medicines, cosmetics or health food.

Hereinafter, the configuration of the present invention will be described in detail.

In order to exert excellent effects when skin regeneration, wrinkle improvement, antioxidant and whitening ingredients are applied to actual skin, they exhibit high activity of skin regeneration, wrinkle improvement, antioxidative and whitening activity at low concentration, Is low in volatility so as to be able to stay for a sufficient time to exhibit skin regeneration, wrinkle improvement, antioxidative and whitening effect, stably maintains the active ingredient on the composition or skin, is easily formulated into medicines and cosmetics, . However, the components satisfying all of the above-mentioned characteristics among the known components are not common. For example, some skin regeneration, wrinkle improvement, antioxidant and whitening ingredients are excellent in skin regeneration, wrinkle improvement, antioxidant and whitening activity even at a low concentration in an in vitro test. However, it's difficult. Other active ingredients have low hydrophilicity and are difficult to formulate into medicines or cosmetics. In addition, some skin regeneration, wrinkle improvement, antioxidant and whitening ingredients may be degraded or transformed into other compounds when exposed to heat, light, or oxygen, which may already have vanished before application to the skin.

As can be seen from the following examples, Eupatilin shows excellent collagen synthesis promoting effect, antioxidative effect and whitening effect at a low concentration. Therefore, it is possible to provide medicines, cosmetics, and cosmetics for skin regeneration, wrinkle improvement, Food and the like.

Accordingly, the present invention provides a pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening comprising a compound represented by the following general formula (1) as an active ingredient.

[Chemical Formula 1]

The compound of Chemical Formula 1 is named 2- (3,4-Dimethoxyphenyl) -5,7-dihydroxy-6-methoxychromen-4-one; It is called 2- (3,4-dimethoxyphenyl) -5,7-dihydroxy-6-methoxy-4H-chromen-4-one and is also called eupatilin.

The compound of formula (1) may be isolated from or synthesized from plant extracts, or a commercially available compound may be used.

The pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening of the present invention may contain glycosides, non-glycosides or pharmaceutically acceptable salts of the compound of formula (1).

The pharmaceutically acceptable salt of the compound of Formula 1 may be an acid addition salt formed using an organic acid or an inorganic acid, and the organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid The organic acid may be selected from the group consisting of hydrochloric acid, hydrobromic acid, hydrochloric acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, , Benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid-based salts, and the inorganic acid includes, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid-based salts. Preferably in the hydrochloride or acetate form, more preferably in the hydrochloride form.

The above-mentioned acid addition salts may be obtained by a) directly mixing the compound of formula 1 and an acid, or b) dissolving and mixing one of them in a solvent or a water solvent, or c) Lt; RTI ID = 0.0 > acid < / RTI > in a solvent and mixing them.

In addition to the above, additionally saltable forms include, but are not limited to, the salts of gabapentin, pregabalin, nicotinate, adipate, hemimarate, cysteine, acetylcysteine, methionine, arginine, Aspartate and the like.

When the compound of Chemical Formula 1 of the present invention is used as a medicine, it may further contain one or more active ingredients showing the same or similar functions. For example, it may contain known skin regeneration, wrinkle improvement, antioxidant and skin whitening ingredients. The skin regeneration, wrinkle improvement, antioxidation and skin whitening effect of the composition of the present invention can be further enhanced by including additional skin regeneration, wrinkle improvement, antioxidant and skin whitening component. When the above ingredients are added, skin safety, easiness of formulation, and stability of effective ingredients can be considered according to the combined use. In one embodiment of the present invention, the composition is a skin regeneration component known in the art comprising retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract, antioxidant components known in the art, such as tocopherol, selenium , Vitamin C and phenolic compounds, whitening ingredients known in the art, substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L- Ascorbic acid, derivatives thereof, and various plant extracts. The additional component may be included in an amount of 0.0001 to 10% by weight based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety, ease of formulation of the compound of Formula 1 .

In addition, the pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation and skin whitening of the present invention may further comprise a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain a variety of ingredients such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, , An extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like.

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

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In the present invention, "skin regeneration effect" refers to restoration of skin tissue against damage caused by external or internal causes of the skin. Damage due to external causes may include ultraviolet rays, external contaminants, wound, trauma, etc. The damage caused by the internal causes may be stress.

In the present invention, the term "wrinkle-reducing effect" refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating wrinkles already generated.

In the present invention, the term "antioxidative effect" refers to the action of free radicals or reactive oxygen species (ROS) due to oxidative stress caused by intracellular metabolism or ultraviolet rays Refers to inhibition of oxidation, and includes removal of free radical or reactive oxygen species, thereby reducing damage to the cells.

In the present invention, the term " whitening effect "refers to not only brightening the skin tone by inhibiting the synthesis of melanin pigment but also improving skin hypercholesterolemia due to ultraviolet rays, hormones or heredity, such as spots or freckles.

The pharmaceutical composition of the present invention can provide a desirable skin regeneration effect, wrinkle improving effect, antioxidative effect and whitening effect when an effective amount of the compound of Formula 1 is included. In the present invention, the term "effective amount" means an amount of a compound capable of promoting regeneration of damaged skin, improving wrinkles, or inhibiting or alleviating oxidation of cells. An effective amount of the compound of formula (1) contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, how the compound is applied to the skin, and the time on the skin. For example, when the composition is commercialized as a pharmaceutical product, the compound of Formula 1 may be contained at a higher concentration than that of a cosmetic product that is routinely applied to skin. Accordingly, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, based on the amount of the compound of formula (1) 6 times a day.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

The present invention can also be provided as a formulation of an external preparation for skin regeneration, wrinkle improvement, antioxidation and whitening comprising the compound of formula (1) as an active ingredient.

When the compound of formula (1) is used as an external preparation for skin, it may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, , Water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for skin And any other ingredients used in the skin sciences. The components can also be introduced in amounts commonly used in the field of dermatology.

When the compound of Formula 1 is provided as an external preparation for skin, it may have a formulation such as, but not limited to, ointments, patches, gels, creams or sprays.

The present invention can also be provided as a cosmetic preparation for skin regeneration, wrinkle improvement, antioxidation and whitening comprising the compound of formula (1) as an active ingredient.

When the compound of Chemical Formula 1 is used as a cosmetic, the cosmetic product containing the compound of Chemical Formula 1 as an active ingredient may be prepared in the form of a general emulsified formulation and a solubilized formulation. For example, creams, essences, cosmetic creams, sprays, gels, packs, sunscreens, make-up bases, liquids such as lotions such as lotion, facial lotion, body lotion, A powder, a cleansing lotion, a makeup removing agent such as a cleansing oil, a cleaning agent such as a cleansing foam, a soap, a body wash and the like.

The cosmetic composition may further contain, in addition to the compound of Formula 1, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, , Ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may contain adjuvants commonly used in the cosmetics field, such as any of the other ingredients.

In the case of a wash-off type cosmetic such as a make-up removing agent, a detergent, etc. in which the active ingredient remains on the skin in a short period of time when the compound of formula (1) is produced into a cosmetic product, . On the other hand, in the case of leave-on type cosmetics, such as lotion, cream, essence, etc., in which the active ingredient remains on the skin for a long period of time, It may be included. In one embodiment of the present invention, although not limited thereto, the composition may contain 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) of the compound of Formula 1 based on the total weight of the composition . When the composition of the present invention contains less than 0.0001% by weight of the compound of formula 1, sufficient skin regeneration, wrinkle improvement, antioxidative and whitening effects can not be expected. When the composition contains more than 10% by weight, This is to prevent a reaction or skin safety problem.

The present invention also relates to a health food for skin regeneration, wrinkle improvement, antioxidant and whitening comprising the compound of formula (1).

Herein, the term "health food" means a food prepared by adding the compound of formula (1) to food materials such as beverage, tea, spice, gum and confectionery, or by encapsulation, powdering or suspension, But it has the advantage that there are no side effects that can occur when a drug is used for a long period of time using a food as a raw material.

Since the health food of the present invention thus obtained can be taken on a daily basis, high skin regeneration, wrinkle improvement, antioxidation and whitening effect can be expected, which is very useful.

When the compound of Chemical Formula 1 is used as a food additive, the compound of Chemical Formula 1 may be added as it is or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Reference Example  1: Eupatilin substance information

2- (3,4-Dimethoxyphenyl) -5,7-dihydroxy-6-methoxychromen-4-one; 2- (3,4-dimethoxyphenyl) -5,7-dihydroxy-6-methoxy-4H-chromen-4-one

CAS No .: 22368-21-4

Where to buy: Tauto Biotech Co., Ltd.

Origin: Artemisia argyi

Example  1: Collagen synthesis effect

The substances of Table 1 below were added to the culture medium of human-derived fibroblasts to examine the promoting effect of collagen synthesis at the cellular level.

The synthesized collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immunoassay Kit). To evaluate the cytotoxicity of human fibroblasts before experimentation at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm of the compound of formula (1) (method of MTT test by culturing fibroblasts (Reference: Mossman T. (1983). Rapid Colorimetric Assay for Cellular Growth & Survival: Application to proliferation & cytotoxicity assays. Journal of Immunological Methods 65, 55-63].

In the experiment, the final concentration of the compound of Chemical Formula 1 was 1 ppm and 10 ppm, and each sample was added to the culture medium of human fibroblasts and cultured for 1 day. Then, the culture broth was taken and the collagen synthesis Was measured at 450 nm using a spectrophotometer. For the comparison of the effects, the degree of collagen synthesis was measured by the same method for the culture medium of fibroblasts (control group) to which nothing had been added and the sample added with vitamin C to a final concentration of 52.8 / / ml. The amount of collagen production was measured as the UV absorbance, and the increase rate of collagen production was calculated as a relative ratio of collagen production to the control. The results are summarized in Table 1 below.

Effect of collagen biosynthesis (number of repeats = 3) sample Absorbance average Collagen growth rate (%) Control group (no addition) 882 - Vitamin C (52.8 ㎍ / ml) 1058 120 Compound of Formula 1 1 ppm 1041 118 Compound of formula 1 0.1 ppm 970 110

As can be seen from the results of Table 1, the collagen synthesis effect was better than that of vitamin C, which is generally known to be capable of collagen synthesis.

Example  2: Collagenase  Activity inhibitory effect

The collagenase inhibitory effect of the substances shown in Table 2 was confirmed as follows.

To evaluate the cytotoxicity of human fibroblasts before experimentation at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm of the compound of formula (1) (method of MTT test by culturing fibroblasts (Reference: Mossman T. (1983). Rapid Colorimetric Assay for Cellular Growth & Survival: Application to proliferation & cytotoxicity assays. Journal of Immunological Methods 65, 55-63.

Human normal skin cells, fibroblasts, were inoculated into 24-well microplates at 2.5 × 10 ⁴ cells per well, cultured in 10% serum DMEM medium and 37 ° C. for 24 hours, and 10% serum DMEM medium was removed The cells were washed once with phosphate buffer solution and incubated for 30 minutes in serum-free DMEM medium supplemented with the compound of formula (I) and in serum-free DMEM medium containing no compound of formula (I).

After 30 minutes of sample treatment, the cells were incubated with 50 ng / ml TNFα (tumor necrosis factor-alpha), a substance known to produce MMP-1, for 24 hours.

In the TNFα-untreated group and the treated group, the TNFα-untreated group was treated as a negative control group and the TNFα-treated treated group as a positive control group.

The supernatant of each well was collected and the amount (ng / ml) of the newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA). The inhibition rate of collagenase activity was determined by MMP -1 production inhibition rate (%) was calculated, and the results are shown in Table 2 below.

The amount of MMP-1 in the control group refers to the amount of MMP-1 in the TNF-treated group, and the amount of MMP-1 in the test group refers to the group to which the substance is added at each concentration.

Inhibitory effect on collagenase activity (number of repeats = 4) sample The amount of MMP-1 (ng / ml) Inhibition rate (%) Compound of Chemical Formula 1 10 ppm 6.0322 61.9347 Compound of Formula 1 1 ppm 17.3627 6.7775 Negative control (TNF untreated) 6.9054 The positive control (TNF treatment) 17.64 -

As can be seen from the results in Table 2, it can be seen that there is an excellent effect on inhibition of collagenase activity.

Example 3: Whitening effect - Confirmation of melanin formation inhibitory effect

The compound of Chemical Formula 1 was added to the culture solution of mouse melanoma cell of B-16 mouse to test the whitening effect at the cellular level. (Lotan R., Lotan D. Cancer Res. 40: 3345-3350, 1980).

To evaluate the toxicity of melanoma cells in rats before the experiment, whitening evaluation was performed by selecting a concentration that is not toxic.

The compound of Chemical Formula 1 was added to the culture medium to a final concentration of 2.5 ug / ml, 5 ug / ml, 10 ug / ml and 20 ug / ml, and the control group albutin was added to the medium to a concentration of 200 ug / And then cultured for 3 days.

Cells were then trypsinized, detached from the culture, centrifuged, and extracted with melanin. The removed cells were melted with 1 ml of sodium hydroxide solution (1N concentration), boiled for 10 minutes, melanin was dissolved, and the amount of melanin produced by measuring the absorbance at 400 nanometers (nm) was measured using a spectrophotometer.

The amount of melanin was measured by an absorbance of 10 ⁶ cells per unit cell, and the amount of melanin produced relative to the control group was calculated as the inhibition rate (%). The results are summarized in Table 3. The experiment was repeated three times.

Inhibitory effect of melanin on cell-level sample Melanin production Inhibition rate (%) Control group (no addition) 0.086 - Control group 1: Arbutin (20 占 퐂 / ml) 0.046 46 The compound of formula (1) (20 占 퐂 / ml) 0.028 68 Compound (1) (10 占 퐂 / ml) 0.054 37 Compound (1) (5 / / ml) 0.071 18 The compound of Formula 1 (2.5 占 퐂 / ml) - -

As can be seen from the results in Table 3, melanoma cells of cultured rats exhibited significantly superior melanin production inhibitory ability even at a much lower concentration than the known whitening substance arbutin (Arbutin).

Example  4: Antioxidant effect - Free radical  Erasure rate

The free radical scavenging activity was measured to confirm the antioxidative activity of the compound of formula (1). Free radical scavenging activity was measured using DPPH. DPPH was purchased from Sigma Co. (Sigma Co., Ltd, USA) and used. First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg / ml) was prepared. Then, ethanol was added to ascorbic acid, which is an antioxidant, as a standard compound and the compound of Formula 1, respectively, to prepare samples at concentrations of 50 μg / ml, 25 μg / ml, 12.5 μg / ml, 6.25 μg / ml and 3.125 μg / ml. Then, the sample and standard DPPH solution were added at the same ratio, stirred well, reacted at 37 ° C for 30 minutes, and absorbance was measured at 520 nm. At this time, ethanol was added instead of the sample to give a control group. IC ₅₀ , which is half maximal inhibitory concentration (inhibition value), was determined for the free radical scavenging ability, and the results are shown in Table 4 below. IC ₅₀ is a common method of expressing free radical scavenging activity with the concentration of ascorbic acid and the compound of formula 1 required to remove 50% of the free radicals of the no-added control group.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (ug / ml; ppm) Ascorbic acid 10 The compound of formula (1) 20

As can be seen from the results in Table 4, the activity is lower than that of ascorbic acid (vitamin C), which is a strong antioxidant, but it is stable as an antioxidant effect with excellent level as a natural substance, Able to know.

Formulation example  1: Preparation of pharmaceutical preparations

1. Manufacturing of powder

0.001 g of the compound of formula (1)

Lactose 1g

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

0.2 mg of the compound of formula (1)

100 mg of corn starch

100 mg of milk

Stearic acid  Magnesium 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of capsules

0.2 mg of the compound of formula (1)

100 mg of corn starch

100 mg of milk

Stearic acid  Magnesium 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

4. Manufacture of rings

0.003 g of the compound of formula (1)

Lactose 1.5 g

Glycerin 1 g

Xylitol  0.5 g

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

5. Manufacture of granules

2 mg of the compound of formula (1)

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60 캜 to form granules, which were then filled in a capsule.

Formulation example  2: Manufacture of cosmetics

1. Manufacture of softening longevity (skin lotion)

As the following composition, the number of softening times containing the compound of formula (1) as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of formula (1)

Beta-1,3-glucan 1.0 wt%

Butylene glycol 2.0 wt%

Propylene glycol 2.0 wt%

Carboxyvinyl polymer 0.1 wt%

0.2% by weight of phage-12 nonylphenyl ether

Polysorbate 80 0.4 wt%

Ethanol 10.0 wt%

0.1% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

2. Manufacture of nutrition lotion (milk lotion)

As in the following composition, a nutritional lotion containing the compound of the formula (1) as an active ingredient was prepared by a conventional method.

0.1% by weight of the compound of formula (1)

Beta-1,3-glucan 1.0 wt%

4.0 wt%

Polysorbate 60 1.5 wt%

1.5% by weight of sorbitan sesquioleate

0.5% by weight liquid paraffin

Caprylic / capric triglyceride 5.0 wt%

Glycerin 3.0 wt%

3.0% by weight of butylene glycol

3.0% by weight of propylene glycol

Carboxyvinyl polymer 0.1 wt%

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

3. Manufacture of nutritional cream

A nutritional cream containing the compound of formula (1) as an active ingredient was prepared according to a conventional method, as shown below.

0.2% by weight of the compound of formula (1)

Beta-1,3-glucan 5.0 wt%

Wax 10.0 wt%

Polysorbate 60 1.5 wt%

≪ tb > < tb > < tb >

0.5% by weight of sorbitan sesquioleate

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

Caprylic / capric triglyceride 5.0 wt%

Glycerin 5.0 wt%

3.0% by weight of butylene glycol

3.0% by weight of propylene glycol

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

4. Manufacture of massage cream

A massage cream containing the compound of formula (I) as an active ingredient was prepared according to a conventional method, as shown below.

0.1% by weight of the compound of formula (1)

Beta-1,3-glucan 3.0 wt%

Wax 10.0 wt%

Polysorbate 60 1.5 wt%

≪ tb > < tb > < tb >

0.8% by weight of sorbitan sesquioleate

Liquid paraffin 40.0 wt%

Squalane 5.0 wt%

Caprylic / capric triglyceride 4.0 wt%

Glycerin 5.0 wt%

3.0% by weight of butylene glycol

3.0% by weight of propylene glycol

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

5. Manufacture of pack

A pack containing the compound of formula (1) as an active ingredient was prepared according to a conventional method, as shown below.

0.2% by weight of the compound of formula (1)

Beta-1,3-glucan 1.0 wt%

Polyvinyl alcohol 13.0 wt%

0.2% by weight of sodium carboxymethylcellulose,

Glycerin 5.0 wt%

Allantoin 0.1 wt%

6.0% by weight of ethanol

0.3% by weight of phage-12 nonylphenyl ether

Polysorbate 60 0.3 wt%

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

Formulation example  3: Topical  Produce

1. Manufacture of gel

As in the following composition, a gel containing the compound of formula (1) as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of formula (1)

0.1% by weight of beta-1,3-glucan

0.05% by weight of sodium ethylenediaminetetraacetate

Glycerin 5.0 wt%

Carboxyvinyl polymer 0.3 wt%

5.0% by weight of ethanol

≪ tb > < tb > < tb >

Triethanolamine 0.3 wt%

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

2. Manufacture of ointment

An ointment containing the compound of formula (I) as an active ingredient was prepared according to a conventional method, as shown below.

0.5% by weight of the compound of formula (1)

Beta-1,3-glucan 10.0 wt%

Wax 10.0 wt%

Polysorbate 60 5.0 wt%

≪ tb > < tb > < tb >

0.5% by weight of sorbitan sesquioleate

Vaseline 5.0 wt%

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

SHARE BUTTER 3.0 wt%

Caprylic / capric triglyceride 5.0 wt%

Glycerin 10.0 wt%

Propylene glycol 10.2 wt%

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

3. Preparation of topical medicament (gel ointment)

A gel ointment agent containing the compound of the formula (1) as an active ingredient was prepared according to a conventional method.

0.5% by weight of the compound of formula (1)

Beta-1,3-glucan 10.0 wt%

1.5% by weight of polyacrylic acid (Carbopol 940)

5.0% by weight of isopropanol

Hexylene glycol 25.0 wt%

Triethanolamine 1.7 wt%

Deionized water to 100%

4. Preparation of topical medicines (patches)

As the following composition, a patch containing a compound of the formula (1) as an active ingredient was prepared by a conventional method.

0.5% by weight of the compound of formula (1)

Beta-1,3-glucan 3.0 wt%

Hexylene glycol 20.0 wt%

Diethylamine 0.7 wt%

1.0% by weight of polyacrylic acid (Carbopol 934P)

0.1% by weight of sodium sulfite

1.0% by weight of polyoxyethylene lauryl ether (E.O. = 9)

1.0% by weight of polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000)

Viscous paraffin oil 2.5 wt%

2.5% by weight of caprylic acid ester / capric acid ester (Cetiol LC)

Polyethylene glycol 400 3.0 wt%

Deionized water to 100%

Formulation example  4: Manufacturing of food

Foods containing the compound of formula (I) of the present invention were prepared as follows.

1. Manufacture of Flour Food

0.05 to 1.0 part by weight of the compound of Formula 1 was added to wheat flour, and a bread, a cake, a cookie, a cracker and a noodle were prepared using the mixture to prepare a health food.

2. Manufacture of dairy products

0.2 part by weight of the compound of Formula 1 was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

3. Manufacturing of wire

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer. The compound of Formula 1 was concentrated under reduced pressure in a vacuum concentrator, sprayed, and dried with a hot-air drier, and the resulting dried product was pulverized to a size of 60 mesh with a pulverizer to obtain a dried powder.

The grains, seeds, and dry powder of the compound of formula (1) prepared above were blended in the following proportions relative to 100 parts by weight of the mixed powder.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

(1) (0.1 part by weight),

(0.5 part by weight),

Rhubarb (0.5 Weight portion )

Formulation example  5: Manufacture of beverages

1. Manufacture of health drinks

0.1 mg of the compound of formula (1)

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water  All 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. Then, the solution thus prepared was filtered and sterilized in a sterilized 2L-container, Lt; RTI ID = 0.0 > health < / 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.

2. Manufacture of vegetable juice

1 g of the compound of formula (I) of the present invention was added to 1,000 mL of tomato or carrot juice to prepare vegetable juice for health promotion.

3. Manufacture of fruit juice

1 g of the compound of formula (1) was added to 1,000 mL of apple or grape juice to prepare fruit juice for health promotion.

Claims (18)

A pharmaceutical composition for skin regeneration and wrinkle improvement comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

A skin external preparation for skin regeneration and wrinkle improvement comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

3. The method of claim 2,
Wherein the external preparation is a formulation of ointment, patch, gel, cream or spray, which is used for skin regeneration and wrinkle improvement.
A cosmetic composition for skin regeneration and wrinkle improvement comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

5. The method of claim 4,
A cosmetic composition for regenerating skin and improving wrinkles, which is a formulation of lotion, essence, lotion, cream, pack, gel, powder, foundation or detergent.
A health food for skin regeneration and wrinkle improvement comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

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