KR102002544B1 - Composition for skin cell regeneration, anti-wrinkle, antioxidant, anti-imflamation, and skin whitening - Google Patents

Abstract

The present invention relates to a composition for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation and skin whitening. The pseudouridine compound according to the present invention promotes the collagen synthesis of the fibroblasts of the skin to suppress the skin regeneration effect, the wrinkle improvement effect and the free radical generation to suppress the antioxidative effect and the NO production to suppress the anti-inflammatory effect and the melanin synthesis, It can be used as a pharmaceutical composition, 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, anti-inflammation, and skin whitening,

The present invention relates to a composition for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation 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, inflammation is an immune response of a human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, active oxygen, free radicals, etc. activate inflammatory factors and cause aging of various diseases and skin. The vasoactive polypeptide, kinin, plasmin and complement, have vasodilatory and contraction and chemotaxis effects, as well as lymphokines such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for inflammation. Arachidonic acid is metabolized through inflammatory mediators such as prostaglandin and lukotrienes via two pathways: cyclooxygenase or lipooxygenase, mediating a variety of inflammatory responses.

On the other hand, in order to eliminate inflammation, elimination of inflammation source, reduction of vital reaction and symptoms is called anti-inflammatory. To date, substances used for antiinflammatory purposes include flutenamic acid, ibuprofen, benzydamine, indomethacin, and steroids, prednisolone, , Dexamethasone, and the like. It is also known that allantoin, azene, hydrocortisone and the like are effective for antiinflammation. However, these materials are limited in their use due to safety of skin, .

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.

Accordingly, it is possible to provide a method for improving the skin regeneration, wrinkle improvement, antioxidant, anti-inflammation and whitening activity, which are more safe and effective in the living body and more effective than the substances having the existing skin regeneration, wrinkle improvement, antioxidant, anti- It is urgently required to develop a component having an amino acid sequence.

Japan Patent No. 8-231370 Japan Patent No. 8-208424 Japan Patent No. 9-40523 Japan Patent No. 10-36283

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

Accordingly, the present inventors found that psoralidin promotes collagen synthesis of fibroblasts in skin and inhibits collagenase activity, thereby promoting skin regeneration, improving wrinkles, and eliminating free radicals, resulting in antioxidative effects, NO production To inhibit the synthesis of melanin and to exhibit 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, antioxidant, anti-inflammation 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, anti-inflammation 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,

The present invention provides a skin external preparation for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation and skin whitening comprising the 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, anti-inflammation 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, antioxidant, anti-inflammation and skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

The pseudouridine according to the present invention promotes collagen synthesis of the fibroblasts of the skin, inhibits collagenase activity, promotes regeneration of the skin, eliminates free radicals to exhibit antioxidative effects, Inhibits the anti-inflammatory effect, suppresses melanin synthesis, and exhibits a whitening effect, so that it can be used in medicines, cosmetics, or health foods.

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

In order to exert excellent effects when skin regeneration, wrinkle improvement, antioxidant, anti-inflammation and whitening ingredients are applied to actual skin, it exhibits high activity of skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening activity at a low concentration, Is low in volatility so as to be able to stay for a sufficient time to exhibit skin regeneration, wrinkle improvement, antioxidant, anti-inflammation and whitening effect, and the active ingredient remains stable on the composition or skin, It is preferable that the formulation is easy, and the skin is safe. 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, anti-inflammatory and whitening ingredients are excellent in skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening activity even at low concentration in in vitro test, It is difficult to apply to real skin. Other active ingredients have low hydrophilicity and are difficult to formulate into medicines or cosmetics. In addition, some skin regrowth, wrinkle improvement, antioxidant, anti-inflammatory and whitening ingredients may be degraded or transformed into other compounds when exposed to heat, light, or oxygen, .

As can be seen in the following examples, pseudouridine exhibits remarkably excellent collagen synthesis promoting effect, antioxidative effect, anti-inflammatory effect and whitening effect at a low concentration, so that it can be used for skin regeneration, wrinkle improvement, antioxidant, anti- Pharmaceuticals, cosmetics, health food, and the like.

Accordingly, the present invention provides a pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation 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 3,9-dihydroxy-2- (3-methylbut-2-enyl) - [l] benzofuro [3,2- c] chromen- 3-methylbut-2-enyl) - [1] benzofuro [3,2-c] chromen-6-one) and is called psoralidin.

The compound of formula (1) may be synthesized or a commercially available compound may be used.

The pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation and skin whitening of the present invention may comprise a pharmaceutically acceptable salt 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 reduction, antioxidant, anti-inflammatory and skin whitening ingredients. The skin regeneration, wrinkle improvement, antioxidant, anti-inflammation and skin whitening effect of the composition of the present invention can be further enhanced by including additional skin regeneration, wrinkle improvement, antioxidant, anti-inflammation 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, anti-inflammation 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. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In the present invention, the term " 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 already-generated wrinkles.

In the present invention, the term 'antioxidative effect' refers to the action of free radicals or reactive oxygen species (ROS), which are highly reactive according 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 'anti-inflammatory effect' refers to inhibition of inflammation. The inflammation is one of defensive responses of biological tissues to certain stimuli, and involves three types of tissue degeneration, circulatory disorder, exudate, and tissue proliferation Complex lesions. More specifically, inflammation is part of congenital immunity and, like in other animals, human congenital immunity recognizes a pattern of cell surfaces that are specifically present in a pathogen. Phagocytes recognize cells with such surfaces as non-magnetic and attack pathogens. If pathogens break through the physical barriers of the body, an inflammatory reaction occurs. Inflammation is a nonspecific defense that creates hostile environments for microorganisms entering the wound. In the inflammatory response, white blood cells that are responsible for the early stage of immune response, when wounded or infected, enter the body to express cytokines. Therefore, the expression level of intracellular cytokine is an index of inflammatory response activation. Examples of skin diseases associated with inflammation include atopic dermatitis, psoriasis, radiation, chemical substances, inflammatory diseases caused by burns, acid burns, vesicular dermatosis, visceral type diseases, itching due to allergies, seborrheic eczema, rose acne, Inflammatory hair loss such as pemphigus vulgaris, polymorphous exudative erythema, erythema nodosum, erythematosus, pharyngitis, alopecia areata, skin T-cell lymphoma, and the like.

In the present invention, the 'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of the 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 reducing effect, antioxidative effect, anti-inflammatory effect and whitening effect when an effective amount of the compound of Chemical 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, inhibiting or alleviating oxidation of cells, suppressing inflammation, or exhibiting a whitening effect . 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 external skin preparation for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation 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 form of cosmetic preparations for skin regeneration, wrinkle improvement, antioxidation, anti-inflammation and whitening, which comprises 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 the formula 1, sufficient skin regeneration, wrinkle improvement, antioxidative, anti-inflammatory and whitening effects can not be expected. To prevent unwanted reactions or skin safety problems.

The present invention also relates to a skin regenerating, wrinkle-improving, antioxidant, anti-inflammatory and whitening health food containing the compound of formula (1).

The term 'health food' as used herein 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, it is very useful because high skin regeneration, wrinkle improvement, antioxidation, anti-inflammation and whitening effect can be expected.

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 by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Reference Example  One: Pthoralidin ( psoralin ) Material Information

3,9-dihydroxy-2- (3-methylbut-2-enyl) - [1] benzofuro [3,2- c] chromen-6-

CAS No .: 18642-23-4

Where to buy: Tauto Biotech Co., Ltd.

Origin and Species: Psoralea corylifolia

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 test substance (method for MTT test by culturing fibroblasts [Reference: Mossman T. ), And the collagen aggregation performance was evaluated by selecting the concentration without cytotoxicity. The results are shown in Table 1.

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.

Collagen biosynthesis effect of compound of formula (1) (number of repeats = 3) sample Absorbance average Collagen growth rate (%) Control group (no addition) 584 - Vitamin C (52.8 ㎍ / ml) 730 125 Compound of Chemical Formula 1 10 ppm 975 167 Compound of Formula 1 1 ppm 777 133

As can be seen from the results of Table 1, the compound of Chemical Formula 1 showed a better collagen synthesis effect at a lower concentration than that of vitamin C, which is generally known to have the ability to synthesize collagen.

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 test substance (method for MTT test by culturing fibroblasts [Reference: Mossman T. ). 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 for 24 hours with 50 ng / ml TNF-alpha (tumor necrosis factor-alpha), a substance known to produce MMP-1.

At this time, the TNF-α-treated group and the treatment group were treated with TNF-α-untreated group and TNF-α-treated group among the control group not containing the compound of formula (1).

The supernatant of each well was collected and the amount (ng / ml) of 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.

[Equation 1]

Inhibitory effect on collagenase activity of the compound of formula (1) (number of repeats = 4) sample The amount of MMP-1 (ng / ml) Inhibition rate (%) Compound of Chemical Formula 1 10 ppm 6.2225 66.2337 Compound of Formula 1 1 ppm 17.7606 6.3909 Negative control (TNF untreated) 6.7609 - The positive control (TNF treatment) 18.1547 -

Example 3 Whitening Effect - Confirmation of Melanogenesis Inhibitory Effect The compound of Chemical Formula 1 was added to the culture medium of mouse melanoma cells (B-16 mouse) to examine 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 5 μg / ml, 10 μg / ml, 20 μg / ml and 200 μg / ml, and the control group, arbutin, B-16 melanoma cells and 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 (Abs.) Inhibition rate (%) Control group (no addition) 0.083 - Control group 1: Arbutin (200 占 퐂 / ml) 0.044 47 The compound of formula (1) (20 占 퐂 / ml) 0.022 73 Compound (1) (10 占 퐂 / ml) 0.039 53 Compound (1) (5 / / ml) 0.056 32

As can be seen from the results of Table 3, the compound of formula (I) has remarkably superior melanin production inhibitory effect on the melanoma cells of the rat cultured as compared with the known whitening substance arbutin (Arbutin).

Example  4: Antioxidant effect - Free radical scavenging 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 a compound of formula (1) to prepare samples at concentrations of 50 / / . 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 the free radical scavenging ability as a 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 8 The compound of formula (1) 16

As shown in Table 4, the compound of the formula (I) is slightly lower in activity than ascorbic acid (Vitamin C), which is a typical strong antioxidant, but is stable as an antioxidative effect substance with a superior level of antioxidative effect as a natural substance. It can be seen that it is suitable.

Example  5: Anti-inflammatory effect - NO  Production inhibitory effect

In order to confirm the antiinflammatory effect and the skin trouble relieving effect of the compound of the formula (1), the nitric oxide (NO) formation inhibition test was carried out by the GRIESS method using the RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, macrophages of mice, were subcultured several times, placed in 24-well plates at 3 × 10 ⁵ per well, and cultured for 24 hours. Subsequently, the compound of Chemical Formula 1 was diluted with the concentrations shown in Table 5 below and replaced with a cell culture medium containing the compound. This concentration means the concentration of the compound dissolved in the medium used for the evaluation, which means 1% = 10 mg / ml, 0.1% = 1 mg / ml, 0.01 = 0.1 mg / ml and 0.001 = 0.01 mg / ml . On the other hand, L-NMMA (L-NG-Monomethylarginine), a NO-production inhibitor, was treated as a positive control and cultured for 30 minutes. Lipopolysaccharide (LPS) 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of the GRIESS solution was added thereto, followed by reaction at room temperature for 10 minutes. Absorbance at 540 nm was measured. The NO inhibitory effect of the compound of formula (1) compared with the negative control treated with LPS only and the NO inhibitory substance was determined, and the results are shown in Table 5 below.

NO generation inhibitory effect sample NO production inhibition rate (%) L-NMMA (positive control, 0.001%) 43.57 Compound (1) (0.001%) 5.63 The compound of formula (1) (0.01%) 25.54 Compound (1) (0.1%) 29.58 Compound (1) (1) 39.42

As shown in Table 5. Compared with L-NMMA, which is a typical anti-inflammatory substance, the compound of Chemical Formula 1 has low activity, but exhibits excellent activity as a natural substance, so that anti-inflammatory effect plays an auxiliary role in whitening and wrinkle improvement, have.

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

2 mg of magnesium stearate

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

2 mg of magnesium stearate

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

Starch 600 mg

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 (6)

A pharmaceutical composition for improving wrinkles comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

A skin external preparation for improving wrinkles 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.
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,
Cosmetics, essences, lotions, creams, packs, gels, powders, foundations or detergents.
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]