KR102138265B1 - Composition for skin cell regeneration, anti-wrinkle, antioxidant, anti-inflammation, and skin whitening comprising 3,23-Diacetyl-N-methylveratramine - Google Patents

Abstract

The present invention provides a composition for skin regeneration, wrinkle improvement, anti-oxidation, anti-inflammatory and whitening, especially for cosmetic, pharmaceutical or health food, comprising diacetylenmethylveratramin or a pharmaceutically acceptable salt thereof. The composition for cosmetic, pharmaceutical or health food of the present invention has few side effects and is safe for the human body, but has excellent skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening effects.

Description

Composition for skin cell regeneration, anti-wrinkle, antioxidant, anti-inflammation, and skin whitening comprising 3,23- Diacetyl-N-methylveratramine}

The present invention relates to a composition related to skin regeneration, wrinkle improvement, anti-oxidation, anti-inflammatory and skin whitening, in particular a composition for cosmetic, pharmaceutical, health food, skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and skin whitening in a small amount. It relates to a composition for a cosmetic, pharmacy, health food excellent in effect.

Collagen is a major matrix protein produced by fibroblasts in the skin. It is present in the extracellular epilepsy, and its important functions are the mechanical firmness of the skin, the resistance of the connective tissue and tissue, the support of cell adhesion, cell division and differentiation (organism Growth or wound healing) is known. It is known that this collagen is reduced by age and photoaging by ultraviolet irradiation, which is closely related to the formation of wrinkles on the skin. In addition, in recent years, as extensive research on skin aging has been developed, important functions of collagen in skin have been revealed.

Active ingredients are known that promote collagen synthesis and show wrinkle improvement effects. For example, retinoic acid, TGF (transforming growth factor), animal placenta-derived protein, betulinic acid, chlorella extract, and the like are known as collagen synthesis promoters. However, the active ingredients have limitations in use due to safety issues such as irritation and redness when applying the skin, or there is a problem that the effect of improving the skin function by actually promoting the collagen synthesis of the skin due to insignificant effect cannot be expected.

On the other hand, free radicals introduced from outside the body or generated in the body cause many problems such as promoting aging of the body or generating cancer. Therefore, many researches and developments have been made on antioxidants that inhibit oxidation by free radicals. Antioxidants are widely distributed in animals and plants, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known for fruits and vegetables. However, the antioxidants existing in nature cannot be expected to have a substantially sufficient effect when applied to the skin. Accordingly, although synthetic antioxidants having excellent antioxidant power and low price are frequently used, their use is limited due to safety concerns such as human side effects.

In addition, inflammation is an immune response of the human body that responds to wounds and diseases, and oxidative stress, such as ultraviolet rays, free radicals, and free radicals, activates inflammatory factors, causing various diseases and skin aging. Vascularly active polypeptides, such as kinin, plasmin, and complement, act to expand and contract blood vessels and chemotaxis. In addition, lymphokas such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to prostaglandin and leukotriene, which are inflammatory mediators, through two pathways: cyclooxygenase or lipooxygenase.

On the other hand, in order to eliminate inflammation, the action of reducing inflammatory sources, reducing biological reactions and symptoms is called anti-inflammatory. Materials that have been used for anti-inflammatory purposes to date include non-steroidal drugs such as flufenamic acid, ibuprofen, benzydamine, and indomethacin, and prednisolone as a steroid system. , Dexamethasone, and the like. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have limitations in the amount of use due to safety problems for the skin, or the effect is insignificant, so that it is not possible to expect an anti-inflammatory effect. There is this.

In addition, it is a constant desire of everyone to have a white skin. It is genetically determined according to the concentration and distribution of melanin in a person's skin, but it is also affected by environmental or physiological conditions such as sun UV rays, fatigue, and stress. Melanin is produced by undergoing a non-enzymatic oxidation reaction after tyrosinase (tyrosinase) acts on tyrosine, an amino acid, to dopa (DOPA) and dopaquinone. The pathway by which melanin is produced is known, but the mechanism by which tyrosinase acts as a precursor to melanin synthesis is still unknown.

Meanwhile, as a general known whitening component, substances that inhibit tyrosinase enzyme activity, such as Kojic acid and Arbutin, hydroquinone, vitamin-C (L-Ascorbic acid), and derivatives thereof and various kinds thereof There are plant extracts. By inhibiting the synthesis of melanin pigments, they can not only realize skin whitening by brightening the skin tone, but also can improve skin hyperpigmentation, such as ultraviolet rays, hormones, or freckles caused by genetics. However, when applied to the skin, there is a problem in that there is a limitation in the amount of use due to safety problems such as irritation and redness, or the effect is insignificant, so that a practical effect cannot be expected.

Therefore, it is safe for the living body, the active ingredient is stable, and above all, it has excellent skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening activity, which is more effective than substances that have existing skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening effects. Development of the ingredients possessed is urgently desired.

Therefore, the present invention solves the above problems, and provides a new active ingredient that has little side effects and is safe for the human body and has excellent skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory, and whitening effects, that is, this useful use of the active ingredient.

In other words, the problem to be solved by the present invention is to provide a composition comprising an active ingredient having excellent efficacy as an active ingredient.

In order to solve the above problems, the present invention is for skin regeneration and wrinkle improvement, antioxidant, including diacetylene methyl veratramine (3,23-Diacetyl-N-methylveratramine) or a pharmaceutically acceptable salt thereof as an active ingredient. Provided is a composition for use in medicine, anti-inflammatory or skin whitening, preferably a cosmetic composition, a pharmaceutical composition, or a composition for health food.

That is, the present invention provides a new use for skin regeneration and wrinkle improvement, anti-oxidation, anti-inflammatory or skin whitening of diacetylenmethylveratramine or a pharmaceutically acceptable salt thereof.

The inventors of the present invention promote the regeneration of the skin and improve wrinkles by diacetylenmethylveratramin (3,23-Diacetyl-N-methylveratramine) promoting collagen synthesis of fibroblasts in the skin and inhibiting collagenase activity. In addition, by scavenging free radicals, the antioxidant effect and NO production were suppressed to have an anti-inflammatory effect, and melanin synthesis was inhibited to show a whitening effect and the present invention was completed.

In the present invention, the term "skin regeneration effect" refers to skin tissue recovery from damage caused by external and internal causes. The damage caused by the external cause may include ultraviolet rays, external contaminants, wounds, trauma, etc., and the damage caused by the internal cause may include stress.

In the present invention, the term "wrinkle improvement effect" refers to suppressing or inhibiting the formation of wrinkles on the skin, or alleviating wrinkles already generated.

In the present invention, the term'antioxidant effect' refers to a cell by a free radical or reactive oxygen species (ROS), which is highly reactive according to oxidative stress due to intracellular metabolism or ultraviolet rays. Refers to inhibiting oxidation, including removing free radicals or free radicals, thereby reducing cell damage.

In the present invention, the term "anti-inflammatory effect" refers to suppressing inflammation, and the inflammation is one of biological tissues' defense responses to certain stimuli, and is associated with three types of tissue degeneration, circulatory disorder and exudation, and tissue proliferation. Complex lesions. More specifically, inflammation is part of innate immunity, and as in other animals, innate immunity in humans recognizes patterns of cell surfaces that are specifically present in pathogens. Phagocytes recognize cells with such a surface as non-magnetic and attack pathogens. If pathogens break through the body's physical barrier, an inflammatory reaction occurs. The inflammatory response is a non-specific defense action that creates a hostile environment against microorganisms that have infiltrated the wound. In an inflammatory reaction, when a wound or an external infectious agent enters the body, the white blood cells responsible for the initial stage immune response flock to express cytokines. Therefore, the amount of cytokine expression in the cells is an indicator of inflammatory response activation. Examples of skin diseases associated with inflammation include atopic dermatitis, psoriasis, radiation, chemicals, erythema diseases triggered by burns, acid burns, blistering skin diseases, thyroid-type diseases, itching due to allergies, seborrheic eczema, rose acne, Inflammatory hair loss such as vulgar erythema, polymorphic exudative erythema, nodular erythema, laryngitis, vulvitis, alopecia areata, skin T-cell lymphoma, but is not limited thereto.

In the present invention, the term'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of melanin pigments, but also improving skin hyperpigmentation such as ultraviolet rays, hormones, or freckles caused by genetics.

Skin regeneration, wrinkle improvement, anti-oxidation, anti-inflammatory and whitening ingredients to show excellent effect when applied to real skin, exhibits high activity skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening activity at low concentrations, and penetrates the skin It has excellent absorption ability, has low volatility to stay for a sufficient time to exhibit skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory, and whitening effects, and keeps the active ingredient stable on the composition or skin, into medicine or cosmetics It is preferred that it is easy to formulate and is also safe for the skin. However, few of the known components satisfying all of the above properties are rare. 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 concentrations in vitro experiments, but have the ability to penetrate and absorb the skin. It is difficult to remove and apply to real skin. Other active ingredients have low hydrophilicity, making them difficult to formulate in medicine or cosmetics. In addition, some skin rejuvenation, wrinkle improvement, antioxidant, anti-inflammatory and whitening ingredients may be decomposed or transformed into other compounds when exposed to heat, light, or oxygen, and the effect may disappear before being applied to the skin. .

The present invention solves the above problems and provides a composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof in order to achieve the object of the present invention.

[Formula 1]

The compound represented by Formula 1 is Acetic acid 2-[1-(3-acetoxy-10,11b-dimethyl-2,3,4,6,6a,11,11a,11b-octahydro-1H-benzo[a] It is called fluoren-9-yl)-ethyl]-1-acetyl-5-methyl-piperidin-3-yl ester, and is also called diacetylenmethylveratramine (3,23-Diacetyl-N-methylveratramine). The present invention is not particularly limited to the method for obtaining the diacetylenmethylveratramine, and chemically synthesized or commercially available materials can be used by methods known in the art.

The "pharmaceutically acceptable salt" of the present invention 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, Malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxy acetic acid, Benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid salts, and inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid salts. It may preferably be in the form of a hydrochloride or acetate, and more preferably in the form of a hydrochloride.

The above-mentioned acid addition salts are either 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-containing solvent, or c) solvent or water washing the compound of Formula 1 It is prepared by the general salt preparation method of placing them in an acid in a solvent and mixing them.

Apart from the above, possible salt forms include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate salt, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt, And aspartates.

The diacetylenmethylveratramins of the present invention may exist in unsolvated forms as well as solvated forms, including forms such as hydrates and ethanolates. The diacetylenmethylveratramin of the present invention may exist in crystalline or amorphous form, and all such physical forms are included in the scope of the present invention.

In addition, when the compound of Formula 1 of the present invention is used as a pharmaceutical, it may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and skin whitening ingredients. If additional skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and skin whitening ingredients are included, the skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and skin whitening effects of the composition of the present invention may be further enhanced. When adding the above components, skin safety, ease of formulation, and stability of active ingredients according to the combined use may be considered. In one embodiment of the present invention, the composition is a skin regeneration or wrinkle improvement ingredient known in the art, including retinoic acid, TGF, animal placenta-derived protein, betulinic acid and chlorella extract; As an antioxidant component known in the art, tocopherol, selenium, vitamin C and phenolic compounds; As a whitening component known in the art, substances that inhibit tyrosinase enzyme activity, such as Kojic acid, Arbutin, hydroquinone, vitamin-C (L-Ascorbic acid); As anti-inflammatory components known in the art, COX-2 inhibitors, prednisolone and allantoin; And one or two or more components selected from the group consisting of these derivatives and various plant extracts. The additional component may be included in an amount of 0.0001% to 10% by weight based on the total composition weight, and the content range may be adjusted according to requirements such as skin safety and ease in formulating the compound of Formula 1 .

In addition, the pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and skin whitening of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain various components such as buffers, sterile water for injection, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and may be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral forms during clinical administration. , Can be prepared using diluents or excipients, such as extenders, binders, wetting agents, disintegrating agents, surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example, starch, calcium carbonate, It can be prepared by mixing sucrose, lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be included.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents, suspension solvents include propylene glycol (Propylene glycol), polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The pharmaceutical composition of the present invention may provide a desirable skin regeneration effect, wrinkle improvement effect, antioxidant effect, anti-inflammatory effect, and whitening effect when an effective amount of the compound of Formula 1 is included. In the present invention, the term'effective amount' refers to an amount of a compound capable of promoting regeneration of damaged skin, improving wrinkles, inhibiting or alleviating oxidation of cells, suppressing inflammation, and showing a whitening effect. . The effective amount of the compound of Formula 1 included in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the compound is applied to the skin, and the length of time it remains on the skin. For example, when the composition is commercialized as a pharmaceutical product, the compound of Formula 1 may be included at a higher concentration than when commercialized as a cosmetic product that is applied to skin on a daily basis. Therefore, the daily dosage is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, more preferably 50 to 60 mg/kg, and 1 to 1 day, based on the amount of the compound of Formula 1 It can be administered 6 times.

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

The present invention can also be provided as a formulation for skin external preparations for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory, 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, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants , Water, ionic or nonionic emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or external preparations for skin It may contain adjuvants commonly used in the field of dermatology such as any other ingredients used as. In addition, the ingredients may be introduced in an amount commonly used in the field of skin science.

When the compound of Formula 1 is provided as an external preparation for skin, it is not limited thereto, and may have a formulation such as ointment, patch, gel, cream or spray.

The present invention can also be provided as a formulation for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory, and whitening cosmetics containing the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as a cosmetic, the cosmetic product containing the compound of Formula 1 as an active ingredient may be prepared in the form of a general emulsifying formulation and a solubilizing formulation. For example, lotion such as soft lotion or nutrition lotion, latex lotion such as facial lotion, body lotion, nutrition cream, moisture cream, cream such as eye cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, liquid It may have a formulation such as foundation type, solid type or spray type, powder, cleansing cream, cleansing lotion, makeup remover such as cleansing oil, cleansing agent such as cleansing foam, soap, body wash, and the like.

In addition, the cosmetic is in addition to the compound of Formula 1, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water , Ionic or nonionic emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the cosmetic field, such as any other ingredient.

The compound of Formula 1 is a relatively high concentration of the compound of Formula 1 in the case of a wash-off type cosmetic such as a makeup remover, detergent, etc., in which the active ingredient stays on the skin within a short period of time when it is commercialized as a cosmetic. It may contain. On the other hand, in the case of a leave-on type cosmetic such as lotion, emulsion, cream, essence, etc., in which the active ingredient stays on the skin for a long period of time, the compound of Formula 1 has a lower concentration than the wash-off type cosmetic. You may include it. Without being limited thereto, in one embodiment of the present invention, the composition may include the compound of Formula 1 in an amount of 0.0001% to 10% by weight (preferably 0.0001% to 1% by weight) based on the total composition weight. Can. 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, antioxidant, anti-inflammatory, and whitening effects cannot be expected, and when it contains more than 10% by weight, allergy This is to prevent unwanted reactions, etc., or skin safety problems.

The present invention also relates to a health food for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening comprising the compound of Formula 1.

In the present specification,'healthy food' is a food prepared by adding the compound of Formula 1 to food materials such as beverages, teas, spices, gums, confectionery, or by encapsulation, powdering, suspension, etc. It means that it has a certain specific effect, but it has the advantage of not having side effects that can occur when taking the drug for a long time using food as a raw material, unlike general drugs.

Since the health food of the present invention obtained in this way can be consumed on a daily basis, high skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory and whitening effects can be expected, and is very useful.

When the compound of Formula 1 is used as a food additive, the compound of Formula 1 may be added as it is or used with other foods or food ingredients, and may be suitably used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, with respect to the raw materials in the production of a food or beverage. However, in the case of long-term intake for health and hygiene purposes or for health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. .

There are no particular restrictions on the type of food. Examples of foods to which the above substances can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, ice creams, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and all of the health foods in the ordinary sense.

The health drink composition of the present invention may contain various flavoring agents or natural carbohydrates, etc., as additional components, like a conventional beverage. The aforementioned 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. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g per 100 mL of the composition of the present invention, preferably about 0.02 to 0.03 g.

In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols , Carbonic acid used in carbonated beverages, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients can be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The composition containing the diacetylenmethylveratramin of the present invention or a pharmaceutically acceptable salt thereof exhibits skin regeneration and wrinkle improvement effects.

The composition containing the diacetylenmethylveratramin of the present invention or a pharmaceutically acceptable salt thereof exhibits an antioxidant effect, an anti-inflammatory effect, or a skin whitening effect.

The composition containing the diacetylenmethylveratramine or the pharmaceutically acceptable salt thereof of the present invention can be used as a safe and excellent cosmetic raw material, pharmaceutical ingredient, and health functional food raw material.

Hereinafter, the following examples and the like will be described to more specifically describe the present invention. However, the embodiments according to the present invention can be modified in many different forms, and the scope of the present invention should not be interpreted as being limited to the embodiments described below. Embodiments of the present invention are provided by way of example to help a specific understanding of the present invention.

Example 1: Collagen synthesis effect

The following Table 1 was added to the culture medium of human-derived fibroblasts to test the effect of promoting collagen synthesis at the cellular level.

The measured collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT). Cytotoxicity was evaluated at human concentrations of 10ppm, 1ppm, 0.1ppm, 0.01ppm, and 0.001ppm of human-derived fibroblasts prior to the experiment (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 concentrations of diacetylenmethylveratramin were 1 ppm and 10 ppm, and each sample was added to the culture medium of human fibroblasts and incubated for 1 day, and then the culture solution was taken at each concentration with the PICP EIA kit. The degree of collagen synthesis was measured at 450 nm using a spectrophotometer. In order to compare the effects, collagen synthesis was measured in the same manner for samples in which fibroblast culture medium (control) and vitamin C added with nothing added to a final concentration of 52.8 µg/ml. The collagen production amount was measured as UV absorbance, and the increase rate of collagen production was calculated as the ratio of collagen production relative to the control group, and the results are summarized in Table 1 below.

Collagen biosynthesis effect of diacetylenmethylveratramin (Repeat number = 3) sample Absorbance average Collagen growth rate (%) Control (no additives) 856 - Vitamin C (52.8 μg/ml) 1070 125 Compound 1 of formula 10 ppm 993 116 1 ppm of compound of formula 1 942 110

As can be seen from the results in Table 1, diacetylenmethylveratramin exhibited excellent collagen synthesis effect at a lower concentration when vitamin C, which is generally known to have collagen synthesis ability, was applied.

Example 2: Collagenaise Active inhibitory effect

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

Cytotoxicity was evaluated at human concentrations of 10ppm, 1ppm, 0.1ppm, 0.01ppm, and 0.001ppm of human-derived fibroblasts prior to the experiment (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], and a collagenase evaluation method was performed by selecting concentrations without cytotoxicity.

Human normal skin cells, fibroblasts, are inoculated into 24-well microplates to be 2.5×10 ⁴ cells per well, incubated in 10% serum DMEM medium and 37° C. for 24 hours, and then 10% serum DMEM medium is removed. And washed once with a phosphate buffer solution, and then incubated for 30 minutes in a serum-free DMEM medium containing diacetylenmethylveratramin and a serum-free DMEM medium containing no diacetylenmethylveratramin as a control.

After 30 minutes of sample treatment, stimulation was performed with 50 ng/ml of TNF-α (tumor necrosis factor-alpha), a substance known to produce MMP-1, and cultured for 24 hours.

At this time, the TNF-α-untreated group and the treated group were treated with TNF-α-treated group and TNF-α-treated group among control groups not containing diacetylenmethylveratramin.

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

The amount of MMP-1 in the control group means the amount of MMP-1 in the TNF-treated group, and the amount of MMP-1 in the experimental group refers to the group in which substances were added at concentrations.

[Equation 1]

Inhibition of collagenase activity of diacetylenmethylveratramin (Repeat number = 4) sample Amount of MMP-1 (ng/ml) Inhibition rate (%) Diacetylenmethylveratramin 10 ppm 6.8008 60.4865 Diacetylenmethylveratramin 1 ppm 16.0396 6.3587 Negative control (TNF-α no treatment) 6.9863 Positive control (TNF-α treatment) 17.1635

Example 3: whitening effect - check the inhibitory effect of melanin production

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

Prior to the experiment, the whitening evaluation was performed by selecting the non-toxic concentration by evaluating the toxicity of the melanoma cells of the mouse.

The compounds of Formula 1 were tested in the culture solution so that the final concentrations were 1 ug/ml, 2.5 ug/ml, 5 ug/ml, 10 ug/ml, and 20 ug/ml, and the control arbutin was added to the medium to be 200 ug/ml, respectively. B-16 melanoma cells were treated and cultured for 3 days.

Thereafter, cells were trypsin-treated, separated from the culture vessel, centrifuged, and melanin was extracted. The detached cells were boiled for 10 minutes by adding 1 ml of sodium hydroxide solution (1N concentration), dissolved melanin, and measured the absorbance at 400 nanometers (nm) using a spectrophotometer to measure the amount of melanin produced.

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

Inhibition effect of melanin production at cell level according to concentration sample Melanin production Inhibition rate (%) Control (no additives) 0.084 - Control 1: Arbutin (1000 mg/ml) 0.030 64 Compound of Formula 1 (20mg/ml) 0.057 32 Compound of Formula 1 (10mg/ml) 0.065 23 Compound of Formula 1 (5mg/ml) - -

As can be seen from the results of Table 3, it can be seen that the compound of Formula 1 has superior melanin production inhibitory ability against cultured rat melanoma cells compared to the known whitening substance Arbutin.

Example 4: Antioxidant effect- Free radical Erasure rate

Free radical scavenging activity was measured to confirm the antioxidant action of diacetylenmethylveratramin. Free radical scavenging activity was measured using DPPH. DPPH was purchased and used by Sigma Co., Ltd. (USA). First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg/ml) was made. Then, ethanol was added to ascorbic acid, an antioxidant, as a compound of Formula 1 and a reference material 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 the 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, instead of the sample, ethanol was added as a control. The free radical scavenging activity was obtained by obtaining the IC ₅₀ , which is the half maximal inhibitory concentration (inhibition median), and the results are shown in Table 4 below. IC ₅₀ is a general method of expressing free radical scavenging ability as a concentration of a compound of formula 1 and ascorbic acid required to remove 50% of free radicals in an additive-free control group.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (ug/ml;ppm) Ascorbic acid 10 Compound of Formula 1 17

As a result of the evaluation, it was found that the activity is lower than that of a representative strong antioxidant substance, ascorbic acid (Vitamin C), but it is stable with an antioxidant effect and is suitable for use as an antioxidant efficacy substance.

Example 5: anti-inflammatory effect- NO Production suppression effect

In order to confirm the anti-inflammatory effect and the skin trouble-relieving effect of the compound of Formula 1, a nitric oxide (NO) formation inhibitory power experiment was conducted by the GRIESS method using a RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are macrophages of mice, were passaged several times, placed in a 24-well plate to enter 3×10 ⁵ cells in one well, and cultured for 24 hours. Subsequently, the compound of Formula 1 was diluted to the concentration shown in Table 5 and replaced with a cell medium containing it. At this time, the NO-producing inhibitor L-NMMA (L-NG-Monomethylarginine) was treated together as a positive control and cultured for 30 minutes, and 1 μg of LPS (Lipopolysaccharide) was treated as a stimulator to incubate for 24 hours. . 100 μl of the supernatant was transferred to a 96-well plate, 100 μl of the GRIESS solution was added, and the mixture was reacted for 10 minutes at room temperature, and the NO inhibitory effect of Compound 1 was determined by measuring the absorbance at 540 nm. It is shown in 5.

NO production suppression effect sample NO production inhibition rate (%) L-NMMA (positive control 0.001%) 26.64 Compound of Formula 1 (0.001 %) 9.80 Compound of Formula 1 (0.01 %) 16.25 Compound of formula 1 (0.1%) 29.47 Compound of formula 1 (1%) 34.60

As a result of the evaluation, the inhibitory effect of NO production was found to be concentration-dependent, and it was judged that an excellent anti-inflammatory effect could be expected.

Formulation example 1: Preparation of pharmaceutical preparations

1. Preparation of powder

Compound of Formula 1 0.001 g Lactose 1 g

A powder was prepared by mixing the above components and filling the gas-tight fabric.

2. Preparation of tablets

Compound of Formula 1 0.2mg Corn starch 100mg Lactose 100mg Magnesium stearate 2mg

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

3. Preparation of capsules

Compound of Formula 1 0.2mg Corn starch 100mg Lactose 100mg Magnesium stearate 2mg

After mixing the above components, the capsules were prepared by filling the gelatin capsules according to a conventional capsule preparation method.

4. Preparation of pills

Compound of Formula 1 0.003 g Lactose 1.5 g glycerin 1 g Xylitol 0.5 g

After mixing the above components, it was prepared to be 4 g per ring according to a conventional method.

5. Preparation of granules

Compound of Formula 1 2 mg Soybean extract 50 mg glucose 200 mg Starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added, dried at 60° C. to form granules, and then packed into the fabric.

Formulation example 2: Preparation of cosmetics

1. Manufacture of flexible cosmetic (skin lotion)

As shown in the following composition, a softening agent containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 1.0 wt% Butylene glycol 2.0% by weight Propylene glycol 2.0% by weight Carboxyvinyl polymer 0.1 wt% PG-12 nonylphenyl ether 0.2% by weight Polysorbate 80 0.4% by weight ethanol 10.0% by weight Triethanolamine 0.1 wt% antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

2. Preparation of nutrient makeup (milk lotion)

As shown in the following composition, a nutrient cosmetic composition containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 1.0 wt% Wax 4.0% by weight Polysorbate 60 1.5% by weight Sorbitan sesquioleate 1.5% by weight Floating paraffin 0.5% by weight Caprylic/Capric Triglyceride 5.0 wt% glycerin 3.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Carboxyvinyl polymer 0.1 wt% Triethanolamine 0.2% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

3. Preparation of nutrition cream

As shown in the following composition, a nutrient cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.2% by weight Beta-1,3-glucan 5.0 wt% Wax 10.0% by weight Polysorbate 60 1.5% by weight Easy Easy 60 Castor Oil 2.0% by weight Sorbitan sesquioleate 0.5% by weight Floating paraffin 10.0% by weight Squalane 5.0 wt% Caprylic/Capric Triglyceride 5.0 wt% glycerin 5.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Triethanolamine 0.2% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

4. Preparation of massage cream

As shown in the following composition, a massage cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 3.0 wt% Wax 10.0% by weight Polysorbate 60 1.5% by weight Easy Easy 60 Castor Oil 2.0% by weight Sorbitan sesquioleate 0.8 wt% Floating paraffin 40.0 wt% Squalane 5.0 wt% Caprylic/Capric Triglyceride 4.0 wt% glycerin 5.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Triethanolamine 0.2% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

5. Manufacture of pack

As shown in the following composition, a pack containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.2% by weight Beta-1,3-glucan 1.0 wt% Polyvinyl alcohol 13.0% by weight Sodium carboxymethylcellulose 0.2% by weight glycerin 5.0 wt% Allantoin 0.1 wt% ethanol 6.0% by weight PG-12 nonylphenyl ether 0.3% by weight Polysorbate 60 0.3% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

Formulation example 3: External preparations for skin Produce

1. Preparation of gel

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

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 0.1 wt% Sodium ethylenediamine acetate 0.05 wt% glycerin 5.0 wt% Easy Easy 60 Castor Oil 0.5% by weight Carboxyvinyl polymer 0.3% by weight ethanol 5.0 wt% Triethanolamine 0.3% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

2. Preparation of ointment

As shown in the following composition, an ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5% by weight Beta-1,3-glucan 10.0% by weight Wax 10.0% by weight Polysorbate 60 5.0 wt% Easy Easy 60 Castor Oil 2.0% by weight Sorbitan sesquioleate 0.5% by weight vaseline 5.0 wt% Floating paraffin 10.0% by weight Squalane 5.0 wt% Shea butter 3.0 wt% Caprylic/Capric Triglyceride 5.0 wt% glycerin 10.0% by weight Propylene glycol 10.2% by weight Triethanolamine 0.2% by weight antiseptic 0.05 wt% Coloring 0.05 wt% Spices 0.05 wt% Purified water to 100% by weight

3. Preparation of topical medication (gel ointment)

As shown in the following composition, a gel ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5% by weight Beta-1,3-glucan 10.0% by weight Polyacrylic acid (Carbopol 940) 1.5 wt% Isopropanol 5.0 wt% Hexylene glycol 25.0 wt% Triethanolamine 1.7 wt% Deionized water to 100% by weight

4. Preparation of pharmaceuticals (patches) for topical administration

As shown in the following composition, a patch containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5% by weight Beta-1,3-glucan 3.0 wt% Hexylene glycol 20.0 wt% Diethylamine 0.7 wt% Polyacrylic acid (Carbopol 934P) 1.0 wt% Sodium sulfite 0.1 wt% Polyoxyethylene lauryl ether (E.O=9) 1.0 wt% Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) 1.0 wt% Viscous paraffin oil 2.5% by weight Caprylic acid ester/capric acid ester (Cetiol LC) 2.5% by weight Polyethylene glycol400 3.0 wt% Deionized water to 100% by weight

Formulation example 4: Preparation of food

Foods containing the compound of Formula 1 of the present invention were prepared as follows.

1. Preparation of flour food

0.05 to 1.0 parts by weight of the compound of Formula 1 was added to wheat flour, and bread, cake, cookie, cracker and noodles were prepared using this mixture to prepare healthy food.

2. Manufacturing of dairy products

0.2 parts 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. Production of wire

The brown rice, barley, glutinous rice, and yulmu were alpha-polished by a known method to distribute the dried one, and then prepared with a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, and then distributed into a powder having a particle size of 60 mesh with a grinder. The compound of Formula 1 was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray or hot air dryer was pulverized to a particle size of 60 mesh to obtain a dry powder.

The dry powders of the above-mentioned grains, seeds, and compounds of formula 1 were blended with 100 parts by weight of the mixed powder to prepare them.

Grains (30 parts by weight of brown rice, 15 parts by weight of barley, 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),
Compound of formula 1 (0.1 parts by weight),
Territory (0.5 parts by weight),
Ground sulfur (0.5 parts by weight)

Formulation example 5: Preparation of beverage

1. Preparation of health drinks

0.1 mg of compound of formula 1

Citric acid 1000 mg

Oligosaccharide 100 g

2 g plum juice

Taurine 1 g

The above ingredients were mixed according to a conventional health drink preparation method, and then purified water was added to prepare a total of 900 ml. After stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered to obtain a sealed 2 L-container and sealed. After sterilization and refrigeration, it was used to prepare the health drink composition of the present invention.

Although the above composition ratio is a mixture of components suitable for a preference drink in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, country of demand, and usage.

2. Preparation of vegetable juice

1 g of the compound of Formula 1 of the present invention was added to 1,000 mL of tomato or carrot juice to prepare a vegetable juice for health promotion.

3. Production of fruit juice

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

Claims (11)

A cosmetic composition for skin regeneration or wrinkle improvement comprising the compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

Antioxidant cosmetic composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

Anti-inflammatory cosmetic composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

delete delete The cosmetic composition according to any one of claims 1 to 3, wherein the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is included in an amount of 0.0001 to 10% by weight based on the total weight of the cosmetic composition. A health food composition for skin regeneration or wrinkle improvement, comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

Antioxidant health food composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

Anti-inflammatory health food composition comprising the compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The health food according to any one of claims 7 to 9, wherein the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is included in an amount of 0.0001 to 10 wt% based on the total weight of the health food composition. Composition. An anti-inflammatory pharmaceutical composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]