KR20120114540A - Cosmetic composition for anti-aging, anti-wrinkle, whitening, anti-inflammatory and anti-biotic effect - Google Patents

Abstract

PURPOSE: A cosmetic composition is provided to enhance anti-oxidant activity effect, MMP-1 generation suppressive effect, collagen synthesis increase effect and skin anti-wrinkling effect. CONSTITUTION: A cosmetic composition for preventing skin aging includes a compound represented by chemical formula 1 as an active ingredient. The cosmetic composition for skin wrinkle improvement includes the compound represented by the chemical formula 1 as the active ingredient. The cosmetic composition for skin whitening includes the compound of the chemical formula 1 as the active ingredient. A cosmetic composition for anti-inflammation includes the compound of the chemical formula 1 as the active ingredient. The cosmetic composition for anti-biotic includes the compound of the chemical formula 1 as the active ingredient. 0.000001-30 wt% of the compound of the chemical formula 1 is included based on the total weight of the cosmetic composition.

Description

Cosmetic composition having anti-aging, anti-wrinkle, whitening, anti-inflammatory and antimicrobial effect {Cosmetic Composition for anti-aging, anti-wrinkle, whitening, anti-inflammatory and anti-biotic effect}

The present invention relates to a cosmetic composition that prevents skin aging, improves skin wrinkles, enhances skin whitening, and exhibits anti-inflammatory and antibacterial effects.

Aging of the skin can be divided into stress, disease, environmental factors, wounds, or internal aging that occurs with age, and light aging caused by exposure to light such as ultraviolet rays.

Aging of the skin is caused by the destruction of the antioxidant protective film present in the living body by the activated free radicals, thereby oxidizing lipids, proteins, polysaccharides and nucleic acids, which are the major constituents of the skin, and thus destroying skin cells and tissues.

In particular, when the protein is oxidized, collagen, hyaluronic acid, elastin, proteoglycan, and fibronectin, which are the connective tissues of the skin, are cleaved, which causes severe over-inflammatory reaction and skin elasticity. If this becomes more severe, mutations in the DNA can lead to mutations, cancer, and reduced immune function.

Therefore, in order to prevent skin aging, it is necessary to prevent skin damage by eliminating free radicals caused by various causes, and also to restore skin by regenerating and proliferating through active metabolism. have.

In addition, skin aging can be promoted by matrix metalloproteinase (MMP, hereinafter referred to as "MMP") which is a collagen degrading enzyme. That is, in vivo, the synthesis and degradation of extracellular matrix such as collagen is properly regulated, but as aging progresses, collagen synthesis decreases and the expression of collagen degrading enzyme MMP is promoted, resulting in decreased skin elasticity and wrinkles. .

Therefore, in order to prevent skin aging and wrinkle generation, it is necessary to control or inhibit MMP expression in which activation is induced in cells.

Furthermore, it is necessary to enhance collagen synthesis in order to improve skin wrinkles.

On the other hand, there is a problem that the skin color is changed independently of skin aging, pigmentation is a major cause for such a change in skin color.

In general, pigments that affect skin color include melanin, melanoids, carotene, hemoglobin, and the most important pigments are melanin.

Melanin plays an important role in preventing skin damage from ultraviolet rays by absorbing or scattering ultraviolet rays, and also excellent in removing free radical species, but sometimes melanin itself generates free radicals, and catechol in melanin structure Or quinones reduce or oxidize other substances, and melanin itself exhibits the properties of free radicals. Therefore, in order to prevent skin discoloration, it is necessary to suppress melanin production.

Melanin is biosynthesized into brown (pheomelanin) and black (eumelanin) polymers through a series of oxidation processes beginning with the conversion of tyrosine, an amino acid, to tyrosinase and conversion to dihydroxy phenylalanine. The biosynthesis process of melanin proceeds in a special type of brown cells in organelles called melanosomes. Through biosynthesis, melanosomes, which contain melanin granules, migrate and accumulate around the nucleus of keratinocytes.

The synthesis of melanin, the number of melanosomes, and the migration to the surrounding keratinocytes are largely genetically influenced, in part, by hormones and ultraviolet rays, and other intracellular regulatory factors, such as cytokines and copper. It is known to be affected by metal ions such as zinc, iron, and interferon, prostaglandin, histamine and the like.

Conventionally, ascorbic acid (JP-A 4-9320), hydroquinone (JP-A-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150), some extracts, etc. It is used as a whitening cosmetic because it has an inhibitory activity of tyrosinase, which is involved in melanin production, but it is used due to problems such as deterioration due to poor stability in cosmetic formulations, coloring, or off-flavor, and unclear efficacy at the biological level. This situation is limited.

It is an object of the present invention to provide a cosmetic composition comprising a new ingredient as an active ingredient that is stable and prevents skin aging, can improve skin wrinkles, and is also effective for skin whitening.

In order to achieve the above object, the present invention provides a cosmetic composition comprising the compound of formula (1) as an active ingredient.

The present invention provides a cosmetic composition for preventing skin aging or improving skin wrinkles comprising the compound of Formula 1 as an active ingredient.

The present invention provides a cosmetic composition for skin whitening comprising the compound of Formula 1 as an active ingredient.

The present invention provides an anti-inflammatory cosmetic composition comprising the compound of Formula 1 as an active ingredient.

The present invention provides a cosmetic composition for antibacterial comprising the compound of Formula 1 as an active ingredient.

The compound of Formula 1 may be included in an amount of 0.000001 to 30.0% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is a formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, cleansing, oils, powder foundations, emulsion foundations, wax foundations, packs, massage creams and sprays. Can be made.

The cosmetic composition according to the present invention has the following advantages by including the compound of formula 1 as an active ingredient.

Cosmetic composition according to the present invention has an excellent antioxidant activity effect, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, and skin wrinkle improvement effect can be used for skin aging prevention or skin wrinkle improvement.

Cosmetic composition according to the invention has a melanin production inhibitory effect can be used for skin whitening purposes.

The cosmetic composition according to the present invention has an inflammatory cytokine expression suppression effect, anti-inflammatory effect and antimicrobial effect by ultraviolet irradiation, and thus can be used for anti-inflammatory and antimicrobial applications.

In addition, the cosmetic composition according to the present invention has an effect of alleviating skin irritation, and has a skin moisturizing effect.

Hereinafter, the present invention will be described in detail.

The present invention provides a cosmetic composition comprising the compound of formula 1 as an active ingredient.

[Formula 1]

The cosmetic composition according to the present invention can be used for anti-aging or skin wrinkle improvement. Specifically, the cosmetic composition according to the present invention, as can be seen in Experimental Examples 1 and 2 to be described later exhibits an antioxidant effect, and as shown in Experimental Example 3 to be described later exhibits an MMP-1 production inhibitory effect, Experimental Example 4 to be described later As can be seen from the collagen synthesis enhances the effect, as can be seen in Experimental Example 11 to be described later because it shows a skin wrinkle improvement effect, it can be usefully used for anti-aging or skin wrinkle improvement.

That is, the cosmetic composition according to the present invention can prevent skin aging by preventing oxidation of skin cells and tissues by scavenging free radicals or free radicals (see Experimental Examples 1 and 2), and also generating MMP-1. By inhibiting the breakdown of collagen to prevent skin elasticity from deteriorating and preventing wrinkles (see Experimental Example 3). Also, by enhancing collagen synthesis, skin elasticity is not reduced and skin wrinkles do not occur. In addition, it is possible to improve the skin wrinkles (see Experimental Example 4), this skin wrinkle improvement effect can be confirmed in Experimental Example 11.

In addition, the cosmetic composition according to the present invention can be used for skin whitening purposes. Specifically, since the cosmetic composition according to the present invention shows a melanin production inhibitory effect, it can be usefully used for skin whitening applications.

That is, the cosmetic composition which concerns on this invention can prevent skin discoloration by suppressing production | generation of the melanin pigment which can cause pigmentation (refer Experimental example 5).

In addition, the cosmetic composition according to the present invention can be used for anti-inflammatory applications. Specifically, the cosmetic composition according to the present invention exhibits an inhibitory effect on inflammatory cytokine expression by ultraviolet irradiation, as can be seen in Experimental Example 6, which will be described later, and as shown in Experimental Example 7, which will be described later, It can be usefully used for anti-inflammatory applications.

Cosmetic compositions according to the invention can be used for antimicrobial purposes. Specifically, since the cosmetic composition according to the present invention exhibits an antimicrobial effect as can be seen in Experimental Example 10, it can be usefully used for antibacterial purposes.

In addition, the cosmetic composition according to the present invention exhibits an effect of alleviating skin irritation as can be seen in Experimental Example 8 described later, and exhibits a skin moisturizing effect as can be seen in Experimental Example 9, and thus, in addition to the specific effects as described above. It has various effects and can be usefully used as a cosmetic.

The compound of Chemical Formula 1 may be separated from the fruit of Popowia cauliflor (popowia cauliflor), bark or fruit of the Lindera erythrocarpa.

That is, a variety of extraction solvents, such as water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol, butanol, and the like), are included in the Popowi caulifolia fruit, the bark bark or the bark tree fruit, Extraction solvent selected from the group consisting of propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof was added to the mixture, followed by stirring under reflux. The process of drying the filtrate under reduced pressure was repeated several times to obtain a concentrate of Poeweia Kaulifola fruit, bark bark or bark tree fruit, which was dissolved in ethanol solution, n-hex, CH ₂ OH ₂ (methylene Lycol), EA (ethyl acetate), n-BuOH, and water layer. The methylene glycol layer obtained by the fractionation was loaded in a column tube filled with silica, and fractionated with n-hexane and ethyl acetate flowing in a gradient of 10: 1 to 1: 1 as a developing solvent. Qualitative analysis by thin membrane chromatography concentrated the fractions up to Rf = 0.43-5.0, and fractions were separated by CHCl ₃ : MeOH (1: 1) on a column packed with Sepadex 20. Then recrystallized with EA and MeOH to obtain a compound of formula 1 of the present invention.

The compound of Formula 1 may be included in an amount of 0.000001 to 30.0% by weight, more preferably 0.0001 to 10.0% by weight, and most preferably 0.001 to 5.0% by weight with respect to the total cosmetic composition.

When the content of the compound of Formula 1 is less than 0.000001% by weight, the antioxidant effect as described above is insignificant and thus cannot be used for skin aging prevention purposes, and when the content of the compound of Formula 1 exceeds 30.0% by weight. E does not show a marked increase in efficacy with increasing content, which may lower economic efficiency.

The cosmetic composition according to the present invention includes components conventionally used in cosmetic compositions in addition to the compound of formula 1 as an active ingredient, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings, and Carrier.

Cosmetic compositions according to the invention can be prepared in any formulations conventionally prepared in the art, for example solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, cleansing, oils, Powder foundations, emulsion foundations, wax foundations and sprays and the like can be formulated, but is not limited thereto.

More specifically, the cosmetic composition according to the present invention may be prepared in the form of a flexible lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder have.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of a spray, additionally chlorofluorohydrocarbon, Propellant such as propane / butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is cleansing, in particular, surfactant-containing cleansing, as a carrier component, an aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosy Nates, fatty acid amide ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Hereinafter, the present invention will be described in more detail with reference to specific production examples. These preparations are merely for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these preparations.

Manufacturing example  1.Isolation and Identification of Compounds of Formula 1

Compound 1 of Formula 1 used in the present invention was isolated and identified by the following method.

5 L of 70% ethanol was added to about 1.0 Kg of the dried Popowia Kauliplea fruit, and the mixture was stirred under reflux and filtered, and the filtrate was dried under reduced pressure. This was repeated twice to obtain about 330 g of concentrate (Pophowia Kauliple Aura crude extract). This was dissolved in a 20% aqueous ethanol solution, and then fractionated into n-hex, CH ₂ OH ₂ (methylene glycol), EA (ethyl acetate), n-BuOH, and water layers to form a methylene glycol layer with silica 230-. It was loaded into a 400 mesh packed column tube and 20 mL was fractionated while flowing n-hexane and ethyl acetate in a gradient of 10: 1 to 1: 1 as a developing solvent. Fractions up to Rf = 0.43-5.0 were concentrated by qualitative analysis with a thin membrane chromatograph, and the fractions were fractionated with CHCl ₃ : MeOH (1: 1) in a column filled with Sepadex 20. After recrystallization with EA and MeOH to obtain about 500 mg. The fractions were analyzed by NMR (and mass). As a result, the component was identified as a compound of Formula 1 (Compound 1).

UV 335, 278 nm, yellow powder FAB-MS m / z : 345 [M + H] +

¹ H-NMR (CDCl ₃ , 300 MHz): 5.46 (1H, dd, J = 12 Hz, 3 Hz), 2.89 (1H, dd, J = 16 Hz, 3 Hz), 3.04 (1H, dd, J = 16 Hz, 12 Hz), 7.41 (5H, m), 4.08 (3H, s), 3.93 (3H, s), 3.87 (3H, s), 3.87 (3H, s).

¹³ C-NMR (CDCl ₃ , 75 MHz): 79.91, 46.33, 190.01, 152.81, 139.16, 153, 83, 141.54, 150.54, 111.09, 138.25, 126.32, 129.17, 128.99, 129.17, 126.32, 62.07, 61.98, 61.84, 61.84.

Experimental Example  One. NBT Measure antioxidant effect

Antioxidant activity was measured by the NBT method to determine the antioxidant effect of the compound of formula 1 isolated according to Preparation Example 1.

In order to measure the antioxidant effect, the active oxygen produced by xanthine and xanthine oxidase was measured by NBT method, and the effect of removing the active oxygen by the sample (compound 1), that is, the active oxygen scavenging effect, was evaluated. . The reactive oxygen scavenging rate is measured by reacting the active oxygen produced by xanthine and xanthine oxidase with Nitro Blue Tetrazolium (NBT) at the wavelength of 560 nm.

The specific measuring method is as follows.

1) To a vial bottle, add 2.4 ml of 0.05M Na ₂ CO ₃ , 0.1ml 3mM xanthine solution, 0.1ml 3mM EDTA solution, 0.1ml BSA solution, and 0.1ml 0.72mM NBT solution After adding 0.1 ml of the solution, it is left at 25 ° C for 10 minutes. Thereafter, 0.1 ml of xanthine oxidase solution is added, stirred quickly, and incubated at 25 ° C for 20 minutes. Thereafter, 0.1 ml of 6 mM CuCl ₂ solution is added to stop the reaction, and the absorbance St at 560 nm is measured.

2) A blank of the sample solution was measured in the same manner as in 1) except that distilled water was used instead of 0.1 ml of xanthine oxidase solution in 1) to measure the absorbance So.

3) In the blank test, absorbance Bt is measured in the same manner as in 1) except that distilled water is used instead of the sample solution in 1).

4) Blank in the blank test was measured in the same manner as in 3) except for using distilled water instead of 0.1 ml of xanthine oxidase solution in 3) to measure the absorbance Bo.

The active oxygen scavenging rate was calculated by the following Equation 1, and the results are shown in Table 1 below.

In Equation 1, St is absorbance at 560 nm after the enzymatic reaction of the sample solution, Bt is absorbance at 560 nm after the enzymatic reaction of the blank test solution, So is absorbance at 560 nm before the reaction of the sample solution without enzyme addition, Bo is the absorbance at 560 nm before the reaction in the absence of enzyme in the blank test solution.

Treatment concentration (ppm) Active oxygen scavenging rate (%) One 36.2 5 54.7 10 75.8 25 85.9 50 91.2

As can be seen in Table 1, the compound of Formula 1 can be seen that the antioxidant power increases in a concentration-dependent manner. In particular, it can be seen that the antioxidant power is 90% or more at 50 ppm or more.

Experimental Example  2. DPPH Measure antioxidant effect

In order to measure the antioxidant effect of the compound of Formula 1 isolated according to Preparation Example 1, the antioxidant activity was measured using the DPPH method.

The DPPH method is to measure antioxidant activity by reducing power using a free group called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The free radical scavenging rate is measured at a wavelength of 560 nm by comparing the degree to which the DPPH is reduced by the sample (compound 1) to reduce the absorbance of the blank.

As a reagent used, a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) was dissolved in methanol to make 100 ml.

The specific measuring method is as follows.

1) Add 0.15 ml of 0.1 mM DPPH solution and 0.15 ml of sample solution to a 96-well plate, stir rapidly, and incubate for 10 minutes at 25 ° C. Thereafter, the absorbance St at 560 nm is measured.

2) The blank of the sample solution was measured in the same manner as in 1) except that methanol was used instead of the 0.1 mM DPPH solution in 1) to measure the absorbance So.

3) In the blank test, absorbance Bt is measured in the same manner as in 1) except that distilled water is used instead of the sample solution in 1).

4) Blank in the blank test was measured in the same manner as in 3) except that methanol was used instead of the 0.1 mM DPPH solution in 3) to measure the absorbance Bo.

The free radical scavenging rate was calculated by the following Equation 2, and the results are shown in Table 2 below.

In Equation 2, St is an absorbance at 560 nm after free radical scavenging of the sample solution, Bt is an absorbance at 560 nm after free radical scavenging of the blank test solution, and So is at 560 nm before reaction when no free radicals are removed from the sample solution. Absorbance, and Bo is the absorbance at 560 nm before the reaction upon free radical addition of the blank test solution.

Treatment concentration (ppm) Free radical scavenging rate (%) One 38.4 2 64.7 5 93.4 10 97.6

As can be seen from Table 2, in the radical scavenging test by DPPH it can be seen that the compound of Formula 1 increases the antioxidant power in a concentration-dependent manner.

Experimental Example  3. MMP -1 suppression effect

The inhibitory effect of MMP-1 production, a collagen degrading enzyme, was measured on the compound of Formula 1 isolated according to Preparation Example 1.

Fibroblasts (Korean Cell Line Bank, South Korea), which are human normal skin cells, were seeded in 48-well microplates (Nunc, Denmark) at 1 × 10 ⁶ cells per well, and treated with DMEM medium (Sigma, United States) and 37 ° C. After incubation for 24 hours in a serum-free DMEM medium containing the compound of Formula 1 added according to Preparation Example 1 and the control was further incubated for 48 hours in a serum-free DMEM medium not containing the compound of Formula 1 as a control. .

After the additional incubation, the supernatant of each well was collected and the amount (ng / ml) of newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA), and MMP- according to Equation 3 below. 1 production inhibition rate was calculated, the results are shown in Table 3.

Treatment concentration (ppm) Inhibition rate of MMP-1 production (%) One 14.21 2.5 18.42 5 29.16 10 54.52 25 75.91

As can be seen in Table 3, it can be seen that the compound of Formula 1 reduces the activity of MMP-1 in a concentration-dependent manner.

Experimental Example  4. Collagen Synthesis Enhancement Effect

Fibroblasts, human normal epithelial cells, were seeded in 48-well microplates at 1 × 10 ⁶ cells per well and incubated in DMEM medium for 24 hours. Thereafter, the mixture was further cultured in serum-free DMEM medium to which the compound of Formula 1 was added according to Preparation Example 1 and serum-free DMEM medium not containing the compound of Formula 1 for 48 hours.

After further incubation, the supernatant of each well was collected and measured in ng / ml by measuring the amount of procollagen (procollagen) type I C-peptide (PICP) using a collagen kit (Takara, Japan). Was measured. Collagen production increase rate was calculated according to the following equation 4, the results are shown in Table 4.

Treatment concentration (ppm) Collagen production increase rate (%) One 4.18 2.5 12.74 5 29.56 10 53.64 25 69.78

As can be seen from Table 4, it can be seen that the collagen production increase rate of the compound of Formula 1 increases in a concentration-dependent manner.

Experimental Example  5. Inhibitory Effects of Melanogenesis Using B16F1 Melanin-forming Cells

In order to confirm the whitening effect of the compound of Formula 1 isolated according to Preparation Example 1, the degree of inhibition of melanin production against B16F1 melanin forming cells was measured to determine the whitening effect.

The B16F1 melanin forming cells used in Experimental Example 5 are cell strains derived from mice, and secrete a black pigment called melanin, which were obtained from ATCC (American Type Culture Collection). During the artificial culture of these cells, samples were treated to compare the degree of reduction of melanin pigment.

The melanin biosynthesis inhibitory effect of B16F1 melanin forming cells was measured as follows.

B16F1 melanin forming cells were divided into 6-well plates at a concentration of 2 X 10 ⁵ per well, and the cells were attached and then incubated for 72 hours by treatment with the compound of Formula 1 isolated in Preparation Example 1 at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, the cell number was measured, and the cells were recovered by centrifugation.

Quantification of intracellular melanin was performed with a slight modification of the method of Rotan: Cancer Res., 40: 3345-3350, 1980. Specifically, the cell pellet was washed once with PBS, and then 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added to vortex for 5 minutes to disrupt the cells. Melt the extracted melanin by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and measure the absorbance of melanin at 405 nm with a microplate reader, and then quantify the melanin. Melanin production inhibition rate (%) was measured.

Melanin production inhibition rate (%) of the B16F1 melanin forming cells was calculated by the following Equation 5, the results are shown in Table 5. IC ₅₀ values in Table 5 refer to concentrations of substances that inhibit melanin production by 50%. At this time, hydroquinone and arbutin, substances known to have melanin production inhibitory effects, were used as positive controls.

In Equation 5, A is the amount of melanin in the well to which the sample is not added, and B is the amount of melanin in the well to which the sample is added.

Name of sample IC ₅₀ Compound 1 0.005% Hydroquinone 0.001% Arbutin 0.02%

As can be seen in Table 5, the compound of Formula 1 significantly inhibits melanogenesis in B16F1 melanin forming cells, and in particular, it is more effective than arbutin used as a positive control and shows a hydroquinone-like effect.

Experimental Example  6. Inhibitory Effects of Ultraviolet Irradiation on Inflammatory Cytokine Expression

With respect to the compound of Formula 1 isolated according to Preparation Example 1, the effect of inhibiting inflammatory cytokine expression expressed by ultraviolet irradiation was evaluated.

Fibroblasts isolated from human epidermal tissue (Fibroblast) were placed in a 24-well test plate each 5 x 10 ⁴ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with UV 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then the PBS was removed and FBS was not added to the cell culture medium (DMEM). Medium) 350 μl was added.

After treating the compound of Formula 1, which was a sample, it was incubated for 5 hours. 150 µl of the culture supernatant was taken to quantify IL-1α to determine the effect of inhibiting inflammatory cytokine expression of the compound of Formula 1. The amount of IL-1α was quantified using an enzyme immunoassay (Enzyme-linked Immunosorbent Assay), and the expression inhibition rate of inflammatory cytokines (IL-1α) was calculated by Equation 6 below, and the results are shown in Table 6. .

In Equation 6, Bo is the amount of IL-1α produced in the wells which have not been irradiated with UV light, and Bt is the amount of IL-1α produced in the wells which have not been irradiated with UV light, and St is the amount of IL-1α IL-1α production of wells.

Treatment concentration (ppm) Inhibitory rate of inflammatory cytokine expression (%) One 5.2 5 11.4 10 31.3 50 61.7 100 73.5

As can be seen from Table 6, the compound of Formula 1 can be seen that inhibit the production of IL-1α, an inflammatory cytokine by ultraviolet light.

Manufacturing example  2. Containing Compound of Formula 1 Cosmetics  Preparation of the composition

In order to measure the effect on the cosmetic composition comprising the compound of formula (1), cosmetics containing the compound of formula (Example 1), glycerin, 1,3 butylene glycol and sorbitol as a moisturizing agent instead of the compound of formula (1) A composition comprising (Comparative Example 1) and a composition not comprising all thereof (Comparative Example 2) were prepared as in Table 7 below. The content units in Table 7 are by weight.

ingredient Example 1 Comparative Example 1 Comparative Example 2 The compound of formula (1) 2.0 - - glycerin 5.0 - 1,3-butylene glycol - 2.5 - Sorbitol - 2.5 - EDTA-2Na 0.02 0.02 0.02 Purified water to 100 to 100 to 100 Cetostearyl alcohol 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 Microcrystallin 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 Trioctanoine 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 BHT 0.05 0.05 0.05 Incense, preservative Suitable amount Suitable amount Suitable amount

Experimental Example  7. Cosmetics  Anti-inflammatory effect of the composition (ear edema experiment)

The anti-inflammatory effect was measured using the cosmetic composition prepared in Preparation Example 2 (Example 1 and Comparative Example 1). Twelve healthy rats were divided into three groups and four rats per sample. First, both ears of rats were wiped well with alcohol (ethanol), and 4 μl of each sample was applied. After 1 hour, 20 μl of acetone was applied to the left ear, and arachidonic acid, an inflammation-causing substance, was applied to the right ear. After 1 hour, the thickness of both ears was repeatedly measured using a micrometer, and the average value was obtained. The anti-inflammatory effect was calculated according to Equation 7 below, and the results are shown in Table 8.

In Equation 7, A is the swelling thickness in the experimental group, B is the swelling thickness in the control group (purified water).

sample Swelling Anti-inflammatory effect (%)
Increased thickness Difference from control
(BA) Control (purified water) 2.84 (B) - Comparative Example 1 2.83 (A) 0.01 0.35 Example 1 1.39 (A) 1.45 51.05

As can be seen in Table 8, it can be seen that the cosmetic composition of Example 1 including the compound of Formula 1 of the present invention shows an excellent anti-inflammatory effect as compared to the cosmetic composition of Comparative Example 1.

Experimental Example  8. Skin irritation test

Samples were prepared by adding lactic acid 0.3% to the cosmetic compositions according to Example 1 and Comparative Example 1 so as to have a pH of 5.5, and then 5 × 20 cm in the lower arm of the arm for 30 males and females. 0.3% lactic acid solution alone and about 0.1 g of the prepared sample were patched for 24 hours, and then removed for 1 hour and 24 hours, and then the skin condition change was visually read according to the following criteria. It was.

The stimulation rate was calculated according to Equation 8 below, and the results are shown in Table 9 below.

Criteria

-: No erythema or unusual symptoms,

+-: Slightly redder than the surroundings,

+: Significantly redder than the surroundings,

++: Reddens and swells more heavily than the surroundings.

sample
Judgment result Stimulation rate
(%) ++ + + - - Example 1 + lactic acid 0.3% - One 2 27 4.44 Comparative Example 1 + lactic acid 0.3% 3 15 11 One 56.66 Lactic acid 0.3% 7 15 8 - 65.55

As can be seen in Table 9, it can be seen that the cosmetic composition of Example 1 according to the present invention shows a large irritation-releasing effect compared to the cosmetic composition of Comparative Example 1 in the irritation-reducing effect by the lactic acid.

Experimental Example  9. Moisturizing effect

Clinical moisturizing effect on the cosmetic composition prepared in Preparation Example 2 (Example 1 and Comparative Examples 1 and 2) was measured as follows.

The questionnaire divided 60 healthy adult men and women who feel dry skin into 3 groups (A, B, C) of 20 randomly. Formulations of Examples 1 and 2 were applied to the face twice daily for 8 weeks. Moisturizing effect was evaluated using Corneometer CM 820 (Corage + Khazaka, Germany) every two weeks from the start of the practical test and after the end, and the results are shown in Table 10 below.

Before use 2 weeks after use 4 weeks after use Example 1 22.4 38.8 45.3 Comparative Example 1 23.6 28.6 30.6 Comparative Example 2 21.7 29.4 30.4

As can be seen in Table 10, it can be seen that the moisturizing effect of the cosmetics containing the compound of Formula 1 (Example 1) is superior to the cosmetics containing the other moisturizing agents (Comparative Examples 1 and 2).

Manufacturing example  3. containing a compound of formula 1 for preservative experiment Cosmetics  Composition manufacturing

As shown in the following Table 11, cosmetics containing the compound of formula 1 (Example 2), cosmetics not containing the compound of formula 1 (Comparative Example 3), and methylparaben as an antiseptic instead of the compound of formula 1 A cosmetic (comparative example 4) containing dazolidinylurethane was prepared. The content units in Table 11 are by weight.

ingredient Example Comparative example 2 3 4 Purified water To 100 To 100 To 100 glycerin 4.0 4.0 4.0 Butylene Glycol 2.0 2.0 2.0 Propylene glycol 2.0 2.0 2.0 EDTA-2Na 0.02 0.02 0.02 Polysorbate 60 1.0 1.0 1.0 Glyceryl stearate 1.5 1.5 1.5 Wax 4.0 4.0 4.0 Macadamia Nut Oil 5.0 5.0 5.0 Squalane 3.0 3.0 3.0 Spices a very small amount a very small amount a very small amount The compound of formula (1) 0.2 - - Methylparaben - - 0.2 Imidazolidinylurea - - 0.2

Experimental Example  10: antimicrobial activity test

The antimicrobial effect on bacteria and fungi was measured using a cosmetic prepared according to Preparation Example 3.

First, in the case of bacteria, 20-30 g of each of the cosmetics of Example 2 and the cosmetics of Comparative Examples 3 and 4 ( E. coli ; ATCC 8739), Pseudomonas aeruginosa (ATCC9027) and Staphylococcus aureus (ATCC6538) were mixed and added so that the initial concentration per sample was 10 ⁷ cfu / g. 1 g of each cosmetic was taken at 1, 7, 14, 21 and 28 days intervals while culturing them in a thermostat at 30-32 ° C for 4 weeks, and the number of viable cells was measured.

Next, in the case of the fungus, Candida albicans (Candida albicans ; ATCC10231), Penicillium cytrinum citrinum ; KCTC2123) and Aspergillus niger (ATCC16404) were added and mixed at 10 ⁷ cfu / g. Occurrence was observed.

The results are shown in Table 12 below.

Cosmetics Bacteria (cfu / g) mold Initial number of bacteria Day 1 Day 7 Day 14 Day 21 Day 28 Example 2 1 x 10 ⁷ 41000 300 200 <100 <100 - Comparative Example 3 1 x 10 ⁷ 54000000 30000000 > 1 x ^{10 8} > 1 x ^{10 8} > 1 x ^{10 8} +++ Comparative Example 4 1 x 10 ⁷ 40000 300 <100 <100 <100 -

Criteria

-: No bad smell, mycelia and spores for 8 weeks

+: Mold develops on walls and lids within 4 weeks

++: Odor and mold on the surface within 4 weeks

+++: Odor and mold on the entire surface within 4 weeks.

As can be seen in Table 12, the cosmetic composition of Comparative Example 3, which does not use a preservative, it can be seen that within four weeks of deodorization and mold on the entire surface, the bacterial bacteria significantly increased even after one day. However, the cosmetic composition of Example 2 containing the compound of formula 1 exhibited a significant antimicrobial effect against bacteria and fungi, 0.2% of the compound of formula 1 (example 2) and conventional preservatives (comparative example 4), respectively And 28 days after the use of 0.4% each, Example 2 using 0.2% of the compound of Formula 1 showed similar preservative strength to Comparative Example 4 using 0.4% of the existing preservative, therefore, the compound of Formula 1 Compared with the existing preservatives, it was found to have much superior antiseptic power.

Experimental Example  11. Skin wrinkle improvement

A clinical experiment was conducted on the skin wrinkle improvement effect of the cosmetic composition containing the compound of formula 1 of the present invention. Clinical trials were evaluated through the actual use test of each cosmetic.

Thirty test subjects (women aged 20 to 35 years) were applied to Example 1 on the right side of the face and Comparative Example 1 on the left side of the face for two consecutive days, two times a day.

After completion of the experiment, the skin wrinkle improvement effect was evaluated by visual observation by an experienced doctor before and after using the product for two months. The experimental results are shown in Table 13 below.

Wrinkle improvement Effective rate (%) Great slightly none Example 1 26 4 0 100% Comparative Example 1 3 6 21 30%

As can be seen in Table 13, the cosmetic composition according to Example 1 of the present invention showed a higher wrinkle improvement effect than the cosmetic composition according to Comparative Example 1. In addition, the skin irritation could not be observed in the skin of the subjects applying the cosmetic composition according to Example 1 of the present invention to the skin.

Claims (7)

A cosmetic composition for preventing skin aging comprising the compound of formula 1 as an active ingredient.
[Formula 1]

A cosmetic composition for improving skin wrinkles comprising the compound of formula 1 below as an active ingredient.
[Formula 1]

Cosmetic composition for skin whitening comprising a compound of formula 1 as an active ingredient.
[Formula 1]

An anti-inflammatory cosmetic composition comprising the compound of formula 1 as an active ingredient.
[Formula 1]

An antimicrobial cosmetic composition comprising the compound of formula 1 as an active ingredient.
[Formula 1]

The method according to any one of claims 1 to 5,
The compound of Formula 1 is a cosmetic composition, characterized in that contained in 0.000001 to 30.0% by weight relative to the total weight of the cosmetic composition. The method according to any one of claims 1 to 5,
The cosmetic composition is a formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, cleansing, oils, powder foundations, emulsion foundations, wax foundations, packs, massage creams and sprays. Cosmetic composition, characterized in that consisting of.