KR102059852B1 - Cosmetic composition containing Oriental Herb Fragrance Active Component Bornyl Acetate for Skin Benefit Ingredient - Google Patents

Abstract

The present invention relates to a composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening.
Carbonyl acetate (bornyl acetate) according to the present invention increases the total amount of collagen of the fibroblasts of the skin to inhibit the skin regeneration effect, wrinkle improvement effect, free radical production to inhibit the antioxidant effect, the production of nitric oxide (NO) anti-inflammatory effect And by reducing the total amount of melanin to show a whitening effect can be used as a pharmaceutical composition, external skin preparations, cosmetic composition or health food.

Description

Cosmetic composition containing Oriental Herb Fragrance Active Component Bornyl Acetate for Skin Benefit Ingredient}

The present invention relates to a composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening.

Collagen is a major substrate protein produced by skin fibroblasts and is present in extracellular epilepsy.Its important functions include mechanical firmness of skin, resistance of connective tissue and tissue binding, support of cell adhesion, cell division and differentiation. Induction of growth or wound healing) is known. This collagen is reduced by age and light aging by ultraviolet irradiation, which is known to be closely related to the wrinkle formation of the skin. In addition, in recent years, extensive research on aging of the skin has been developed an important function of collagen in the skin.

Active ingredients are known that promote collagen synthesis and exhibit wrinkle improvement. For example, retinoic acid, transforming growth factor (TGF) [non-patent document 1], protein derived from animal placenta [patent document 1], betulinic acid (patent document 2), and chlorella extract [ Patent Documents 3 and 4 are known as collagen synthesis promoting substances. However, the active ingredients are limited in the amount of use due to safety problems such as irritation and redness when applying the skin, or there is a problem in that the effect of improving the skin function is not expected by substantially promoting collagen synthesis of the skin.

On the other hand, free radicals introduced from outside the living body or generated in the living body cause many problems such as promoting aging of the living body or generating cancer. Therefore, many researches and developments on antioxidants that inhibit oxidation by active oxygen have been made. Antioxidants are widely distributed in copper and plant systems, and phenolic compounds, flavonoids, tocopherols, vitamin C, selenium, and the like, which are widely contained in fruits and vegetables, are known. However, the antioxidants present in nature are not expected to have a substantial enough effect on skin application. Therefore, many synthetic antioxidants having excellent antioxidant power and low cost are used, but their use is limited due to safety concerns such as human side effects.

In addition, inflammation is an immune response of the human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activates inflammatory factors to cause various diseases and aging of the skin. Vascular activating polypeptides, such as kinin, plasmin or complement, act as vasodilation, contraction and chemotaxis, as well as lymphokahs such as interleukin-6 (IL-6). Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to prostaglandin or leukotrienes, which are inflammatory mediators, through two pathways, cyclooxygenase or lipooxygenase, to mediate various inflammatory reactions.

On the other hand, the anti-inflammatory agent that acts to remove the inflammatory source, reduce the biological response and symptoms to eliminate inflammation. The substances used for anti-inflammatory purposes so far are nonsteroidal flufenamic acid, ibuprofen, benzydamine or indomethacin, and prednisolone as a steroid system. Or dexamethasone. In addition, allantoin, azene or hydrocortisone are known to be effective in anti-inflammatory, but these substances have a problem in that they are limited in the amount of use due to the safety of the skin, or the effects are not so small that substantially no anti-inflammatory effect can be expected. have.

In addition, it is a general desire to have white and fair skin. The color or brightness of the skin is genetically determined by the concentration and distribution of melanin in human skin, but is also affected by environmental or physiological conditions such as solar ultraviolet light, fatigue or stress. Melanin is a type of amino acid tyrosine (tyrosine) enzymes tyrosinase (tyrosinase) acts as a catalyst to be sequentially converted to dopa (DOPA) and dopaquinone (dopaquinone) and then made through a non-enzymatic oxidation reaction. As such, the pathway through which melanin is made is known, but it is still unknown in detail what causes the tyrosinase to be triggered in the mechanism that induces melanin synthesis.

On the other hand, as a known whitening component, substances that inhibit tyrosinase enzyme activity such as kojic acid or arbutin, hydroquinone, vitamin C (L-Ascorbic acid) or derivatives thereof and various There is a plant extract. By inhibiting the synthesis of melanin pigments, they not only brighten the skin tone to realize skin whitening, but also improve skin hyperpigmentation such as blemishes and freckles due to ultraviolet rays, hormones or heredity. However, when the skin is applied, there are limitations on the amount of use due to safety problems such as irritation and redness, or there is a problem in that a substantial effect cannot be expected due to a slight effect.

Therefore, it is safe for the living body, the active ingredient is stable, and above all, the skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening activity is superior to the existing skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening substance. There is an urgent need for the development of these components.

[Prior literature]

[Patent Documents]

(Patent Document 0001) Japanese Patent Laid-Open No. 8-231370

(Patent Document 0002) Japanese Patent Application Laid-Open No. 8-208424

(Patent Document 0003) Japanese Patent Laid-Open No. 9-40523

(Patent Document 0004) Japanese Patent Laid-Open No. 10-36283

[Non-Patent Documents]

(Non-Patent Document 0001) Cardinale G. et al, Adv. Enzymol., 41, p. 425, 1974

Thus, the inventors of the present invention increase the total amount of collagen of the fibroblasts of the skin to promote skin regeneration, improve wrinkles, eliminate free radicals to show the antioxidant effect, and produce nitric oxide (NO) production Inhibition shows an anti-inflammatory effect, and the total amount of melanin is markedly reduced to show a whitening effect, thus completing the present invention.

Accordingly, an object of the present invention to provide a composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the following formula (1) as an active ingredient:

[Formula 1]

As a means for solving the above problems, the present invention

It provides a pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the formula (1) as an active ingredient.

 [Formula 1]

As another means for solving the above problems, the present invention

It provides a skin external preparation for skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

As another means for solving the above problems, the present invention

It provides a cosmetic composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the formula (1) as an active ingredient.

[Formula 1]

As another means for solving the above problems, the present invention

It provides a health food for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

Carbonyl acetate (bornyl acetate) according to the present invention enhances the total amount of collagen of the fibroblasts of the skin, promotes the regeneration of the skin, exhibits an antioxidant effect by eliminating free radicals, producing nitric oxide (NO) involved in the inflammatory response By inhibiting the anti-inflammatory effect is excellent, it can be used in medicine, cosmetics or health food by showing a whitening effect by significantly reducing the total amount of melanin.

EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.

Skin regeneration, wrinkle improvement, anti-oxidation, anti-inflammatory or whitening ingredients have excellent skin regeneration, anti-wrinkle, anti-oxidation, anti-inflammatory or whitening activity at low concentration. It is preferable that the ability to absorb is excellent. In addition, it has low volatility so that it can stay for a sufficient time for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or whitening effect, the active ingredient remains stable on the composition or skin, and is easily formulated into medicine or cosmetics, It is also desirable to be safe for the skin. However, among the known components, components satisfying all of the above properties are rare. For example, some skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening ingredients have excellent skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening activity at low concentrations in vitro, but have the ability to penetrate and absorb skin. It is difficult to apply to real skin apart. Other active ingredients are less hydrophilic and difficult to formulate in medicine or cosmetics. In addition, some skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening ingredients may decompose or transform into other compounds when exposed to heat, light, or oxygen, and the effects are already lost before application to the skin. .

As can be seen in the following examples, the carbonyl acetate (bornyl acetate) has an effect of significantly increasing the total amount of collagen, significantly reduced the total amount of melanin, excellent free radical scavenging rate, inhibits the production of nitric oxide It can be used as an active ingredient in medicines, cosmetics or health foods for regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening.

Accordingly, the present invention provides a pharmaceutical composition for skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or skin whitening comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

The compound of Formula 1 is called a bornyl acetate, and may include all possible isomers.

The compound of Formula 1 may be synthesized or used a commercially available compound.

The pharmaceutical composition for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or skin whitening of the present invention may include 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, 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, aminooxyoxyacetic acid, Benzenesulfonic acid, p-toluenesulfonic acid or methanesulfonic acid salts. Inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid or boric acid salts. It may preferably be in the form of hydrochloride or acetate, more preferably in the form of hydrochloride.

The above-mentioned acid addition salts may be a) directly mixing the compound of formula 1 and an acid, b) dissolving and mixing one of them in a solvent or a hydrous solvent, or c) solvent or submerging the compound of formula 1 It is prepared by a general salt preparation method which is placed in an acid in a solvent and mixed with them.

In addition to the above, salts that can be additionally salted are gava salt, gabapentin salt, pregabalin salt, nicotinate, adipate salt, hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or Aspartate.

In addition, when using the compound of Formula 1 of the present invention as a medicine, it may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or skin lightening ingredients. Inclusion of additional skin rejuvenation, anti-wrinkle, antioxidant, anti-inflammatory or skin whitening ingredients may further enhance the skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or skin lightening effects of the compositions of the present invention. When the ingredient is added, skin safety, ease of formulation, and stability of the active ingredients may be considered. In one embodiment of the invention, the composition is a skin regeneration component known in the art, Retinoic acid, TGF, protein from animal placenta, betulinic acid or chlorella extract; Antioxidant ingredients known in the art include tocopherol, selenium, vitamin C or phenolic compounds; As a whitening component known in the art, a substance that inhibits tyrosinase enzyme activity such as Kojiic acid or Arbutin, Hydroquinone, L-Ascorbic acid, derivatives thereof, and It may further comprise one or two or more components selected from the group consisting of various plant extracts. Additional components may be included in at least 0.0001% and up to 10% by weight relative to the total composition weight. For example, at least 0.0001 wt% to at most 1 wt%, at least 0.0001 wt% to at most 0.1 wt%, at least 0.0001 wt% to 0.001 wt%, at least 0.001 wt% to 10 wt%, at least 0.001 wt% to 1 It may be up to 0.001% by weight or less, up to 0.1% by weight, 0.01% by weight to 10% by weight, 0.01% by weight to 1% by weight or less. The content range may be adjusted according to requirements such as skin safety, ease of formulating the compound of Formula 1, and the like.

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

Pharmaceutically acceptable carriers may contain various components such as buffers, sterile water for injection, general saline or phosphate buffered saline, sucrose, histidine, polysorbate, and the like.

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, in various dosage forms, orally or parenterally, in the case of clinical administration. It may be formulated using diluents or excipients such as extenders, binders, wetting agents, disintegrants or surfactants.

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

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use include suspensions, solvents, emulsions, or syrups, and may include various excipients, for example, wetting agents, sweeteners, fragrances, or preservatives, in addition to water or liquid paraffin, which are commonly used simple diluents.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations or suppositories. Non-aqueous or suspending solvents include vegetable oils such as propylene glycol, polyethylene glycol or olive oil; Injectable esters such as ethyloleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter or glycerogelatin may be used.

In the present invention, the "skin regeneration effect" refers to the recovery of skin tissue against damage caused by external and internal causes of the skin. The damage caused by the external cause may include ultraviolet rays, external pollutants, wounds or traumas, and the damage caused by the internal cause may include stress.

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

In the present invention, the term "antioxidative effect" refers to a cell caused by free radicals or reactive oxygen species (ROS) that are highly reactive due to oxidative stress caused by intracellular metabolism or ultraviolet light. Refers to the inhibition of oxidation, including the removal of free radicals or reactive oxygen species, thereby reducing damage to cells.

In the present invention, the "anti-inflammatory effect" refers to inhibiting inflammation. Inflammation is one of biological tissue's defense response to a certain stimulus and refers to a complex lesion involving three kinds of tissue degeneration, circulatory disorder and exudation and tissue proliferation. More specifically, inflammation is part of innate immunity and, as in other animals, human innate immunity recognizes patterns of cell surfaces that are specific to pathogens. Phagocytes recognize cells with such surfaces as nonmagnetic devices and attack pathogens. If the pathogen breaks through the body's physical barrier, an inflammatory reaction occurs. Inflammatory reactions are nonspecific defenses that create a hostile environment against invading microbes. In inflammatory reactions, when wounded or an external infectious agent enters the body, white blood cells in the early stages of immune response rush to express cytokines. Therefore, the expression level of cytokines in cells is an indicator of inflammatory response activation. Examples of skin diseases associated with inflammation include atopic dermatitis, psoriasis, radiation, chemicals or burns, and erythematous diseases; Itching due to acid burns, bullous skin disease, thyroid disease or allergies; Inflammatory hair loss, such as seborrheic eczema, rose acne, vulgaris, polymorphic exudative erythema, nodular erythema, glansitis, vulvitis or alopecia areata; Cutaneous T-cell lymphomas, and the like.

In the present invention, the "whitening effect" means not only brightening skin tone by inhibiting the synthesis of melanin pigments, but also improving skin hyperpigmentation such as blemishes and freckles due to ultraviolet rays, hormones or heredity.

The pharmaceutical composition of the present invention may provide a desirable skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or whitening effect when including an effective amount of the compound of formula (1). 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, inhibiting inflammation, or exhibiting 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 by which the compound is applied to the skin, or the time it stays on the skin. For example, when the composition is manufactured as a medicine, it may include the compound of Formula 1 in a higher concentration than when manufactured as a cosmetic that is routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, 1 to 1 day, based on the amount of the compound of Formula 1 It may be administered six times.

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

The present invention can also be provided as a formulation of a skin external preparation for skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or 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 is further added to fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants. Skin external preparations such as water, ionic emulsifiers, nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic active agents, lipophilic active agents or lipid vesicles It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in the art. The ingredients may also be introduced in amounts generally used in the field of dermatology.

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

The present invention can also be provided in the formulation of a cosmetic for skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening comprising the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as a cosmetic, a cosmetic prepared by containing the compound of Formula 1 as an active ingredient may be prepared in the form of a general emulsion formulation or solubilized formulation. Lotion such as, for example, a flexible lotion or a nourishing lotion; Latex, such as facial lotion or body lotion; Creams such as nourishing creams, moisture creams, eye creams and the like; essence; Cosmetic ointments; spray; Gels; pack; Sunscreen; Makeup base; Foundations such as liquid type, solid type or spray type; powder; Makeup removers such as cleansing creams, cleansing lotions, cleansing oils; Formulations such as cleansing foams, soaps, cleaning agents such as body washes, and the like.

In addition to the compound of Formula 1, the cosmetics may contain fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, Typically used in cosmetics such as ionic emulsifiers, nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic actives, lipophilic actives or lipid vesicles It may contain an adjuvant used.

The compound of Formula 1 may be a relatively high concentration of the compound of Formula 1 in the case of a wash-off type of cosmetics such as a makeup remover or a detergent in which an active ingredient stays on the skin within a short time when it is commercialized. It may include. On the other hand, in the case of a leave-on type cosmetic such as a lotion, an emulsion, a cream or an essence in which the active ingredient stays on the skin for a long time, the compound of Chemical Formula 1 has a lower concentration than the wash-off type cosmetic. It will be okay. Although not 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 weight of the composition. Can be. When the composition of the present invention comprises less than 0.0001% by weight of the compound of Formula 1, sufficient skin regeneration, anti-wrinkle, antioxidant, anti-inflammatory or whitening effect is not expected, and when it contains more than 10% by weight allergy This is to prevent unwanted reactions or skin safety problems may occur.

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

In the present specification, "health food" is a food prepared by adding the compound of Formula 1 to food materials, such as beverages, teas, spices, gums, confectionery, encapsulated, powdered, suspensions, etc. It means to bring a specific effect, but unlike the general medicine has the advantage that there is no side effect that can occur when taking long-term use of the drug as a raw material.

Since the health food of the present invention thus obtained can be consumed on a daily basis, a high skin regeneration, wrinkle improvement, antioxidant, anti-inflammatory or whitening effect 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 appropriately used according to a conventional method. The mixed amount of the active ingredient can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials. However, when ingested for a long time for the purpose of health and hygiene, it may include the content 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 is no particular limitation on the kind of food. Examples of the food to which the substance may be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages or vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the usual beverage. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin, cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumartin, stevia extract; Synthetic sweeteners such as saccharin and aspartame; The proportion of said 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 is various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, Glycerin, alcohol or carbonation agent, and the like. In addition, the health food of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage, or vegetable beverage. These components can 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 merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Reference Example  One: Bornyl acetate substance  Information

Bicyclo [2.2.1] heptan-2-ol, 1,7,7-trimethyl-, acetate, (1S-endo)-(Bicyclo [2.2.1] heptan-2-ol, 1,7,7-trimethyl -, acetate, (1S-endo)-)

CAS No .: 5655-61-8

Where to buy: sigma-aldrich korea

Example  1: collagen total increase effect

The following Table 1 was added to the culture of human-derived fibroblasts to measure the total amount of collagen at the cellular level. The total amount of collagen was measured using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT). Evaluation of cytotoxicity at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm in human-derived fibroblasts before the experiment (method of culturing fibroblasts and performing MTT test [Reference: Mossman T. (1983) Rapid Colorimetric Assay for Cellular Growth & Survival: application to proliferation & cytotoxicity assays.Journal of Immunological Methods 65, 55-63].

In the experiment, the final concentration of the compound of Formula 1 was set to 1 ppm and 10 ppm, respectively, and each sample was added to the culture medium of human fibroblasts and cultured for 1 day, and then the culture medium was taken and collagen at each concentration was collected using PICP EIA kit. The total amount was measured at 450 nm using a spectrophotometer. For comparison of the effects, the total amount of collagen was measured in the same manner with respect to the fibroblast culture medium (control) and the sample to which vitamin C was added at a final concentration of 52.8 ppm. The total amount of collagen was measured as UV absorbance, and the increase rate of the total amount of collagen was calculated as the ratio of total amount of collagen relative to the control group, and the results are summarized in Table 1 below.

Increase effect of total collagen (repeated number = 3) sample Absorbance average Collagen Growth Rate (%) Compound of Formula 1
(1 ppm) 478 104 Compound of Formula 1
(10 ppm) 504 110 Control
(No addition) 456 - Vitamin c
(52.8 ppm) 552 121

As can be seen from the results of Table 1, the compound of Formula 1 (10 ppm) is lower than the case of applying vitamin C (52.8ppm) known to have collagen synthesis ability, but excellent collagen synthesis ability as a natural substance Stable with Therefore, it can be seen that the compound of Formula 1 can be used for skin regeneration and wrinkle improvement.

Example  2: melanin total reduction effect

The whitening effect was examined by adding to the culture medium of B-16 mouse melanoma cells and measuring the total amount of melanin at the cellular level (Lotan R., Lotan D. Cancer Res. 40: 3345-3350 , 1980). Before the experiment, rat melanoma cells were assessed for toxicity, and non-toxic concentrations were selected for whitening evaluation.

The final concentration of the compound of Formula 1 was 5 ppm, 10 ppm, 20 ppm in the culture medium, and arbutin, a control group, was added to the medium so as to be 200 ppm, and treated with B-16 melanoma cells, respectively, and cultured for 3 days.

Thereafter, cells were trypsinized, removed from the culture vessel, and centrifuged to extract melanin. The detached cells were added with 1 ml of sodium hydroxide solution (1 N concentration), boiled for 10 minutes to dissolve the melanin, and the absorbance was measured at 400 nanometers (nm) using a spectrophotometer to measure the amount of melanin produced.

The melanin amount was measured by the absorbance of the unit cell count (10 ⁶ cells), the total amount of melanin relative to the control group was calculated as the inhibition rate (%) and the results are summarized in Table 2. The experiment was repeated three times.

Reduction effect of total melanin at the cellular level with different concentrations sample Melanin % Inhibition Control group (no addition) 0.071 - Control 1: Arbutin (200 ppm) 0.039 45 Compound of Formula 1 (20 ppm) 0.047 33 Compound of Formula 1 (10 ppm) 0.052 27 Compound of Formula 1 (5 ppm) 0.068 4

As shown in Table 2, the compound of the formula (1) can be used for whitening by a similar reduction in the total amount of melanin for the melanoma cells of rats cultured at low concentrations compared to arbutin (200 ppm), which is known as a whitening substance. And it was found.

Example  3: antioxidant effect Free radical  Erasure rate

Free radical scavenging activity was measured to confirm antioxidant activity. Free radical scavenging activity was measured using DPPH. DPPH was purchased from Sigma Co., Ltd, USA. First, a standard DPPH ethanol solution at 1.5 mM (0.06 mg / mL) concentration was made. Ethanol was added to the ascorbic acid as an antioxidant as a compound of Formula 1 and a reference material, respectively, to prepare samples at concentrations of 50 ppm, 25 ppm, 12.5 ppm, 6.25 ppm, and 3.125 ppm. Then, the sample and the standard DPPH solution were added in the same ratio, stirred well, and then reacted at 37 ° C. for 30 minutes, and the absorbance was measured at 520 nm. At this time, ethanol was added instead of the sample as a control. The free radical scavenging ability was obtained from IC ₅₀ , which is a half maximal inhibitory concentration, and the results are shown in Table 3 below. IC ₅₀ is a general method of expressing free radical scavenging ability as a concentration of ascorbic acid and a compound of formula (1) required to remove 50% of free radicals from an additive-free control group.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (ppm) Ascorbic acid 8 Compound of Formula 1 19

As can be seen in Table 3, the compound of Formula 1 is low in activity compared to ascorbic acid (Vitamin C), which is a representative strong antioxidant, but is used as an antioxidant effect material because it is stable with an excellent level of antioxidant effect as a natural substance. It is understood that the following is suitable.

Example  4: anti-inflammatory effect NO  Generation suppression effect

In order to confirm the anti-inflammatory effect and skin trouble relief effect, nitric oxide (NO) formation inhibitory experiment was performed by GRIESS method using 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 for 3 × 10 ^{5 in} one well, and then cultured for 24 hours. Subsequently, the cells of formula 1 were diluted and replaced with cell media containing 1%, 0.1%, 0.01%, 0.001%. At this time, 0.001% of L-NMMA (L-NG-Monomethylarginine), a NO-producing inhibitor, was treated with a positive control and incubated for 30 minutes, and cultured for 24 hours by treating 1 μg of LPS (Lipopolysaccharide) as a stimulant. It was. 100 μl of the supernatant was transferred to a 96-well plate, 100 μl of GRIESS solution was added thereto, reacted at room temperature for 10 minutes, and the absorbance at 540 nm was measured to determine the NO inhibitory effect of the compound of Formula 1, and the result It is shown in Table 4 below.

NO production inhibitory effect sample NO production inhibition rate (%) L-NMMA (positive control, 0.001%) 40.33 Compound of Formula 1 (0.001%) 0.26 Compound of Formula 1 (0.01%) -2.08 Compound of Formula 1 (0.1%) -2.57 Compound of Formula 1 (1%) 19.92

As can be seen from Table 4, the compound of Formula 1 shows a low activity but excellent activity as a natural substance, compared to the representative anti-inflammatory drug L-NMMA, the anti-inflammatory effect is a secondary role in the whitening or wrinkle improvement It can act as a synergistic effect.

Formulation example  1: Preparation of Pharmaceutical Formulations

1. Preparation of powder

0.001 g of compound of Formula 1

1g lactose

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

0.2 mg of compound of Formula 1

Corn Starch 100mg

Lactose 100mg

Stearic acid  Magnesium 2mg

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

3. Preparation of Capsule

0.2 mg of compound of Formula 1

Corn Starch 100mg

Lactose 100mg

Stearic acid  Magnesium 2mg

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

4. Manufacture of rings

0.003 g of compound of Formula 1

Lactose 1.5 g

1 g of glycerin

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

2 mg of compound of formula 1

Soybean Extract 50mg

Glucose 200 mg

Starch 600 mg

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

Formulation example  2: manufacture of cosmetics

1. Preparation of Soft Cosmetics (Skin Lotion)

Like the following composition, a flexible longevity containing the compound of formula 1 as an active ingredient was prepared according to a conventional method.

0.1 wt% of a compound of Formula 1

Beta-1,3-Glucan 1.0 wt%

Butylene Glycol 2.0 wt%

Propylene Glycol 2.0 wt%

Carboxy vinyl polymer 0.1 wt%

Fiji-12 nonylphenyl ether 0.2% by weight

Polysorbate 80 0.4 wt%

Ethanol 10.0 wt%

0.1% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

2. Preparation of Nutrients (Milk Lotion)

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

0.1 wt% of a compound of Formula 1

Beta-1,3-Glucan 1.0 wt%

Beeswax 4.0 wt%

Polysorbate60 1.5 wt%

Sorbanthesquioleate 1.5 wt%

0.5% by weight of liquid paraffin

Caprylic / Capric Triglycerides 5.0 wt%

Glycerin 3.0 wt%

Butylene Glycol 3.0 wt%

Propylene Glycol 3.0 wt%

Carboxy vinyl polymer 0.1 wt%

0.2% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

3. Preparation of nutrition cream

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

0.2 wt% of a compound of Formula 1

Beta-1,3-Glucan 5.0 wt%

Beeswax 10.0 wt%

Polysorbate60 1.5 wt%

Sebum 60 Cured Castor Oil 2.0 wt%

0.5 wt% sorbitan sesquioleate

10.0% by weight of liquid paraffin

Squalane 5.0 wt%

Caprylic / Capric Triglycerides 5.0 wt%

Glycerin 5.0 wt%

Butylene Glycol 3.0 wt%

Propylene Glycol 3.0 wt%

0.2% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

4. Preparation of Massage Cream

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

0.1 wt% of a compound of Formula 1

Beta-1,3-Glucan 3.0 wt%

Beeswax 10.0 wt%

Polysorbate60 1.5 wt%

Sebum 60 Cured Castor Oil 2.0 wt%

Sorbanthesquioleate 0.8 wt%

40.0% by weight of liquid paraffin

Squalane 5.0 wt%

Caprylic / Capric Triglyceride 4.0 wt%

Glycerin 5.0 wt%

Butylene Glycol 3.0 wt%

Propylene Glycol 3.0 wt%

0.2% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

5. Manufacture of pack

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

0.2 wt% of a compound of Formula 1

Beta-1,3-Glucan 1.0 wt%

Polyvinyl Alcohol 13.0 wt%

Sodium Carboxymethyl Cellulose 0.2% by weight

Glycerin 5.0 wt%

Allantoin 0.1 wt%

Ethanol 6.0 wt%

0.3% by weight of sebum-12 nonylphenyl ether

Polysorbate 60 0.3 wt%

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

Formulation example  3: External skin preparations  Produce

1. Preparation 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 wt% of a compound of Formula 1

Beta-1,3-Glucan 0.1 wt%

0.05% by weight of ethylenediamine sodium acetate

Glycerin 5.0 wt%

Carboxy vinyl polymer 0.3 wt%

Ethanol 5.0 wt%

Fiji-60 Cured Castor Oil 0.5 wt%

0.3% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

2. Manufacture of ointments

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.

0.5% by weight of compound of formula 1

Beta-1,3-Glucan 10.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 5.0 wt%

Sebum 60 Cured Castor Oil 2.0 wt%

Sorbanthesquioleate 0.5 wt%

Vaseline 5.0 wt%

10.0% by weight of liquid paraffin

Squalane 5.0 wt%

Shea Butter 3.0 wt%

Caprylic / Capric Triglycerides 5.0 wt%

Glycerin 10.0 wt%

Propylene Glycol 10.2 wt%

0.2% by weight of triethanolamine

0.05 wt% preservative

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100% by weight

3. Preparation of Topical Drug (Gel Ointment)

As the following composition, a gel ointment comprising the compound of formula 1 as an active ingredient was prepared according to a conventional method.

0.5% by weight of compound of formula 1

Beta-1,3-Glucan 10.0 wt%

1.5% by weight of polyacrylic acid (Carbopol 940)

Isopropanol 5.0 wt%

Hexylene glycol 25.0 wt%

Triethanolamine 1.7 wt%

Deionized water to 100% by weight

4. Preparation of medicament (patch) for topical administration

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

0.5% by weight of 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)

Sodium sulfite 0.1 wt%

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

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

Viscous paraffin oil 2.5% by weight

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

Polyethylene glycol 400 3.0 wt%

Deionized water to 100% by weight

Formulation example  4: manufacture of food

Food containing the compound of formula 1 of the present invention was 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 the flour, and using the mixture to prepare bread, cake, cookies, crackers and noodles to prepare a health 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. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh. Black beans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then roasted to prepare a powder having a particle size of 60 mesh. The compound of Chemical Formula 1 was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The dry powders of the grains, seeds and the compound of formula 1 prepared above were prepared by blending the powders in 100 parts by weight in the following ratio.

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

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

Compound of formula 1 (0.1 parts by weight),

Ganoderma lucidum (0.5 parts by weight),

Foxglove (0.5 Parts by weight )

Formulation example  5: manufacture of beverages

1. Manufacture of health drinks

0.1 mg of compound of Formula 1

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Purified water  By adding a full 900 mL

After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained by sterilization in a sterilized 2L-container, sealed sterilized and stored in the refrigerator It was used to prepare a healthy beverage composition of the invention.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and intended use.

2. Preparation of Vegetable Juice

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

3. Preparation 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 (24)

A pharmaceutical composition for improving skin regeneration or wrinkles comprising the compound represented by Formula 1 as an active ingredient:
[Formula 1]

Skin external preparation for skin regeneration or wrinkle improvement comprising the compound represented by Formula 1 as an active ingredient:
[Formula 1]

The method of claim 2,
The external preparation is an external preparation for skin regeneration or wrinkle improvement in the form of an ointment, patch, gel, cream or spray.
Cosmetic composition for skin regeneration or wrinkle improvement comprising the compound represented by the formula (1) as an active ingredient:
[Formula 1]

The method of claim 4, wherein
Cosmetic composition for skin regeneration or wrinkle improvement which is a formulation of a lotion, essence, lotion, cream, pack, gel, powder, foundation or detergent.
Health food for skin regeneration or wrinkle improvement comprising the compound represented by the following formula 1 as an active ingredient:
[Formula 1]

delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete