KR101264003B1 - 3′,4′,5,6,7-Pentahydroxyflavanone compound and Method for manufacturing the same, Cosmetic composition produced therefrom - Google Patents

Abstract

The present invention has anti-inflammatory and antibacterial properties, can prevent skin aging or wrinkles, and also has a skin whitening effect, 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound and a method for preparing the same, prepared from the same It relates to a cosmetic composition.

Description

3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound and preparation method thereof, and cosmetic composition prepared therefrom {3 ′, 4 ′, 5,6,7-Pentahydroxyflavanone compound and Method for manufacturing the same , Cosmetic composition produced therefrom}

The present invention relates to a 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound and a method for preparing the same, and a cosmetic composition prepared therefrom. More specifically, the present invention has anti-inflammatory and antibacterial properties and prevents skin aging. And it relates to a 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound and a method for producing the same, and a cosmetic composition prepared therefrom that can enhance skin whitening.

Aging of the skin can occur by activating free radicals, which destroys the antioxidant protective film present in the living body, thereby destroying skin cells and tissues by oxidizing lipids, proteins, polysaccharides and nucleic acids, which are the major constituents of the skin.

In order to prevent such skin aging, it is necessary to prevent skin damage by eliminating free radicals caused by various causes and to restore skin by regenerating and proliferating already damaged cells through active metabolism.

In addition, skin aging may be promoted by matrix metalloproteinase (hereinafter referred to as 'MMP') which is a collagen degrading enzyme. In other words, 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 wrinkles, it is necessary to regulate MMP expression or inhibit its activity in cells.

On the other hand, there is a problem that the skin color changes independently of skin aging, the main cause of the skin color change is pigmentation. In general, pigments affecting skin color include melanin, melanoids, carotene, hemoglobin, etc. Of these, melanin generates free radicals on its own and reduces or oxidizes other substances by catechol or quinone in the melanin structure and It also indicates the nature of free radicals. Therefore, in order to prevent skin discoloration, it is necessary to suppress melanin production.

Accordingly, the development of new materials to prevent skin aging and improve skin whitening is urgently needed.

The present invention is to solve the above problems, 3 ', 4', 5, 6, 7-pentahydroxy flavanone compound that has anti-inflammatory and antibacterial properties, can prevent skin aging or wrinkles, and also have a skin whitening effect And it aims at providing the manufacturing method and the cosmetic composition manufactured from this.

As one aspect for achieving the above object, the present invention provides a 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound represented by the following formula.

[Chemical Formula]

 As another aspect, the present invention, the skin composition for preventing skin aging or wrinkle improvement or skin whitening or anti-inflammatory or antibacterial comprising the 3 ', 4', 5, 6, 7-pentahydroxy flavanone as an active ingredient To provide.

As another aspect, the present invention, the process for preparing compound 1 by reacting 1,3,5,6-pentahydroxybenzene and caffeic acid as shown in the following scheme; Cyclizing the compound 1 to produce the following compound 2; And it provides a method for producing a 3 ', 4', 5, 6, 7-pentahydroxyflavanone (pentahydroxyflavanone) compound comprising the step of reducing the compound 2 to produce the compound 3.

[Reaction Scheme]

The present invention has the following effects.

First, the 3 ', 4', 5,6,7-pentahydroxyflavanone compound according to the present invention and the cosmetic composition comprising the same as an active ingredient inhibits the production of MMP-1 and promotes the synthesis of collagen by aging the skin. Or it can prevent skin wrinkles.

Second, the 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound and the cosmetic composition comprising the same as an active ingredient according to the present invention has a skin whitening effect because it can inhibit melanin production.

Third, the 3 ', 4', 5,6,7-pentahydroxyflavanone compound according to the present invention and the cosmetic composition comprising the same as an active ingredient, anti-inflammatory effect and mold and by inhibiting the inflammatory cytokine expression by ultraviolet irradiation It has an antibacterial effect against bacteria.

1 is a graph showing the free radical scavenging rate (%) according to the concentration of 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound in the experiment of free radical scavenging rate of Experimental Example 2.

Hereinafter, the present invention will be described in detail.

First, the 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound of the present invention will be described in detail.

The 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound of the present invention is represented by the following formula.

[Chemical Formula]

The 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound of the present invention may be used for anti-aging or anti-wrinkle purposes.

Specifically, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound of the present invention may be used for skin aging prevention purposes as it shows antioxidant effects as described in Experimental Examples 1 and 2 described below. As shown in Experimental Example 3 and Experimental Example 4 to be described later, it can be used for skin wrinkle improvement as it shows an effect of inhibiting MMP-1 production and enhancing collagen synthesis.

That is, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, as shown in Experimental Examples 1 and 2, prevents oxidation of skin cells and tissues by scavenging free radicals or free radicals. Therefore, skin aging can be prevented. In addition, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, as shown in Experiment 3, prevents degradation of collagen by inhibiting MMP-1 production, thereby reducing skin elasticity and skin Wrinkles can be prevented. In addition, the 3 ', 4', 5, 6, 7-pentahydroxy flavanone compound, as shown in Experimental Example 4, it is possible to prevent the decrease in skin elasticity and skin wrinkles by promoting collagen synthesis.

The 3 ', 4', 5,6,7-pentahydroxyflavanone compound of the present invention can be used for skin whitening applications.

Specifically, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, as can be seen in Experimental Example 5 to be described later, may be usefully used for skin whitening by inhibiting melanin production.

That is, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, as shown in Experimental Example 5, can prevent skin discoloration by inhibiting the production of melanin pigment which may cause pigmentation. have.

The 3 ', 4', 5,6,7-pentahydroxyflavanone compounds of the present invention can be used for anti-inflammatory applications.

Specifically, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound may exhibit an anti-inflammatory effect by inhibiting inflammatory cytokine expression by ultraviolet irradiation, as shown in Experimental Example 6. It can be usefully used for anti-inflammatory applications.

The 3 ', 4', 5,6,7-pentahydroxyflavanone compounds of the present invention can be used for antimicrobial applications.

Specifically, the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound is the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound as shown in Experimental Example 10. It can be seen that there is an antimicrobial effect indirectly from the cosmetic containing as an active ingredient exhibits an antimicrobial effect.

Next, the manufacturing method of the 3 ', 4', 5, 6, 7- pentahydroxy flavanone compound of this invention is demonstrated in detail.

First, compound 1 is obtained by reacting 1,3,5,6-pentahydroxybenzene and caffeic acid as shown in the following scheme. In this case, the compound 1 may be obtained by the reaction at high temperature after the addition of the BF ₃ Et ₂ O catalyst.

Subsequently, the compound 1 obtained by the following reaction formula is heated, and the cycloaddition reaction is carried out, and the compound 2 is obtained. In this case, Compound 2 may be obtained by using DMSO as a solvent and reacting at high temperature after adding iodine as a catalyst.

Subsequently, compound 2 obtained in the following scheme is reduced to synthesize compound 3, that is, 3 ', 4', 5, 6, 7-pentahydroxyflavanone. In this case, the compound 3 may be obtained by reacting hydrogen using a palladium catalyst.

[Reaction Scheme]

Next, the cosmetic composition of the present invention will be described in detail.

The cosmetic composition includes the aforementioned 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound as an active ingredient.

The 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound may be included in an amount of 0.000001 to 30.0 wt% based on the total cosmetic composition, preferably 0.0001 to 10.0 wt%, and more preferably. Preferably 0.001 to 5.0% by weight. When the content of 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone is less than 0.000001% by weight, the above-described efficacy may be insignificantly expressed, and the 3 ′, 4 ′, 5,6 When the content of, 7-pentahydroxyflavanone is more than 30.0% by weight, the economical efficiency may be lowered because the efficacy is not significantly improved.

The cosmetic composition according to the present invention includes the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound having an anti-aging or skin wrinkle improvement or skin whitening or anti-inflammatory or antibacterial effect as an active ingredient. It will have the above efficacy.

The cosmetic composition according to the present invention includes components commonly used in cosmetic compositions in addition to the active ingredient 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, for example, antioxidants, stabilizers, And conventional adjuvants and carriers such as solubilizers, vitamins, pigments and flavorings.

The cosmetic composition includes the aforementioned 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound, which has an effect on skin aging and wrinkle improvement, skin whitening effect, and anti-inflammatory and antibacterial effect. .

For example, the cosmetic composition, as can be seen in Experimental Example 7, because it can exhibit an anti-inflammatory effect by inhibiting the inflammatory cytokine expression by ultraviolet irradiation can be usefully used for skin anti-inflammatory applications. In addition, the cosmetic composition, as can be seen in Experimental Example 10, because it exhibits an antimicrobial effect can be usefully used for antibacterial purposes.

In addition to the above-described efficacy, the cosmetic composition, as can be seen in Experimental Example 8 to be described later, has an effect of alleviating skin irritation, and also as described in Experimental Example 9 to be described later, has a moisturizing effect.

As such, the cosmetic composition may be used as a useful cosmetic material because it possesses various effects such as skin moisturization in addition to specific effects such as the above-described anti-aging.

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

Specifically, the cosmetic composition 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.

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

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

When the formulation is a solution or emulsion, a carrier, solubilizer or emulsion may be used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, Fatty acid esters of 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan are used.

When the formulation is a suspension, the carrier component may be 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, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation is surfactant-containing cleansing, the carrier component may be aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether. Sulfates, alkylamidobetaines, aliphatic 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: 3 ′, 4 ′, 5,6,7- Pentahydroxyflavanone  Compound manufacturing

As in the following scheme, Compound 1 was obtained by reacting 1,3,5,6-pentahydroxybenzene and caffeic acid. Thereafter, DMSO solvent and an iodine catalyst were added to the compound 1 thus obtained, followed by cycloaddition reaction at a high temperature to synthesize compound 2. Thereafter, hydrogen was added to the compound 2 obtained under a palladium catalyst to reduce the reaction to synthesize compound 3.

Compound 3 thus obtained was analyzed for structure using NMR and Mass analyzer. Compound 3 was identified as 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone as shown below.

yellow powder ESI-MS m / z : 305 [M + H] + 1H-NMR (MeOD - d 4, 400 MHz): 6.97 (1H, d, J = 1.6 Hz, H-2 '), 6.85 (1H, dd, J = 1.6 Hz, H-6 '), 6.79 (1H, d, J = 8.0 Hz H-5'), 5.94 (1H, s, H-8), 5.33 (1H, dd, J = 12.4, 2.8 Hz, H-2), 3.13 (1H, doublet of doublets, J = 12.4, 17.2 Hz, H-3), 2.74 (1H, doublet of doublets, J = 17.2, 3.2 Hz, H-3) 13C-NMR (MeOD - d 4 ,, 100 MHz): 79.77 (C-2), 43.04 (C-3), 196.78 (C-4), 149.23 (C-5), 125.57 (C-6), 156.84 (C-7), 95.44 (C-8), 156.36 ( C-9), 101.97 (C-10), 130.56 (C-1 '), 113.81 (C-2'), 145.30 (C-3 '), 145.77 (C-4'), 115.05 (C-5 ' ), 118.39 (C-6 ')

Experimental Example  One. NBT Antioxidant effect measurement

The antioxidant effect of the 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound prepared according to Preparation Example 1 was measured using the NBT method.

That is, the antioxidant effect of the 3 ', 4', 5,6,7-pentahydroxyflavanone compound is the amount of free radicals produced by xanthine and xanthine oxidase and 3 ', 4', 5 of the present invention. The amount of active oxygen removed by the, 6,7-pentahydroxyflavanone compound was measured through the active oxygen scavenging ratio obtained by the NBT method. This active oxygen scavenging rate was obtained by measuring the absorbance at 560 nm of the blue color produced by reacting the active oxygen produced by xanthine and xanthine oxidase with Nitro Blue Tetrazolium (NBT).

The specific measuring method is as follows.

1) To a vial bottle was added 2.4 ml of 0.05M Na ₂ CO ₃ , 0.1 ml of 3mM xanthine solution, 0.1ml of 3mM EDTA solution, 0.1ml of BSA solution, and 0.1ml of 0.72mM NBT solution, where 3 0.1 ml of a ', 4', 5, 6, 7-pentahydroxyflavanone compound sample is added, and the mixture is left at 25 ° C. for 10 minutes. Then, 0.1 ml of xanthine oxidase solution is added, stirred at high speed, 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) Blank of the sample is obtained by measuring the absorbance (So) in the same manner as in 1) except that distilled water is used instead of 0.1 ml of xanthine oxidase solution in 1).

3) The blank test is obtained by measuring the absorbance (Bt) in the same manner as in 1) except that distilled water is used instead of the sample in 1).

4) Blank in blank test is obtained by measuring the absorbance (Bo) in the same manner as in 3) except that distilled water is used instead of 0.1 ml of xanthine oxidase solution in 3).

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

[Equation 1]

Active oxygen scavenging rate (%) = [1- (St-So) / (Bt-Bo)] × 100

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

Treatment concentration (ppm) Active oxygen scavenging rate (%) One 48.2 2 66.3 5 92.2 10 94.7 20 97.8

As can be seen in Table 1, the 3 ', 4', 5, 6, 7-penta hydroxyflavanone compound of the present footing increased the antioxidant power in a concentration-dependent manner. In particular, when the concentration of 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound was 5 ppm or more, the antioxidant power was 90% or more.

Experimental Example  2 : DPPH Measurement of antioxidant activity

Antioxidant activity of 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound prepared according to Preparation Example 1 was measured using the DPPH method.

This DPPH method is to measure the antioxidant activity by reducing power using free groups of DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. Antioxidant activity of the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound sample was reduced by DPPH by 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound sample. The free radical scavenging rate was obtained by measuring the absorbance at a wavelength of 560 nm compared with the absorbance of the blank test solution.

In this case, 61.88 mg of DPPH free radical (Aldrich Chem. Co., MW = 618.76, 0.1 mM solution) 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 to a 96-well plate, stir at high speed and incubate for 10 minutes at 25 ° C. Thereafter, the absorbance St 'is measured at a wavelength of 560 nm.

2) The blank of the sample is measured for absorbance (So ') in the same manner as in 1) except that methanol is used instead of 0.1mM DPPH solution in 1).

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 in 1).

4) Blank in blank test measures the absorbance (Bo ') in the same manner as in 3), except that methanol was used instead of the 0.1 mM DPPH solution in 3).

The free radical scavenging rate was calculated by the following equation.

&Quot; (2) "

% Free radical scavenging rate = [1- (St'-So ') / (Bt'-Bo')] × 100

As can be seen in Figure 1, in the free radical scavenging test by DPPH 3 ', 4', 5,6,7-pentahydroxyflavanone compound of the present invention increased the antioxidant power in a concentration-dependent, concentration of 5 ppm At least 90% of free radicals were removed.

Experimental Example  3: MMP -1 suppression effect

The inhibitory effect of MMP-1 production, a collagen degrading enzyme, of 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound prepared according to Preparation Example 1 was measured by the following method.

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 at a temperature of 37 ° C. and DMEM medium (Sigma, USA). After 24 hours of incubation, serum-free DMEM medium to which the 3 ', 4', 5,6,7-pentahydroxyflavanone compound sample was added to the experimental group and serum-free DMEM medium not containing the sample as a control group Incubated further for 48 hours.

The supernatants of each of the wells incubated were collected, and the amount of MMP-1 (ng / ml) of the experimental group and the control group was measured using an MMP-1 assay kit (Amersham, USA), and MMP according to Equation 3 below. The inhibition of -1 production was calculated and the results are shown in Table 2.

&Quot; (3) "

% Inhibition of MMP-1 production = [1- (amount of MNP-1 in the experimental group / amount of MMP-1 in the control group)] × 100

Treatment concentration (ppm) Inhibition rate of MMP-1 production (%) One 5.61 2.5 20.10 5 47.81 10 76.40 25 87.88

As can be seen in Table 2 above, the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound of the present invention reduced the activity of MMP-1 in a concentration-dependent manner.

Experimental Example  4: collagen synthesis promoting effect

The collagen synthesis enhancing effect of the 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound prepared according to Preparation Example 1 was confirmed through the following method.

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 sample was added to the serum-free DMEM medium to which the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound sample was added and the serum-free DMEM medium containing no sample to the control group for 48 hours. Incubated with

The supernatant of each additionally cultured well was collected and measured by the collagen kit (Takara, Japan) to measure the amount of procollagen (procollagen) type IC-peptide (PICP) and converted to ng / ml. Was measured.

The collagen production increase rate was calculated according to Equation 4 below, the results are shown in Table 3.

&Quot; (4) "

Collagen production increase rate (%) = [(collage amount of experimental group / collagen amount of control group) -1] × 100

Treatment concentration (ppm) Collagen production increase rate (%) One 3.2 2.5 44.6 5 64.8 10 110.5 25 147.6

As can be seen in Table 3 above, the increase rate of collagen production of the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound of the present invention increased in a concentration-dependent manner.

Experimental Example  5: Effect of Inhibiting Melanogenesis Using B16F1 Melanin-forming Cells

The skin whitening effect of 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound prepared according to Preparation Example 1 was determined by measuring the degree of melanin production inhibition on B16F1 melanin forming cells.

The B16F1 melanin forming cell used in Experimental Example 5 is a cell strain derived from a mouse, and secreted a black pigment called melanin, which was distributed from ATCC (American Type Culture Collection). The melanin production inhibitory effect was confirmed by comparing and evaluating the degree of melanin reduction by treating the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound samples during artificial culture of the cells.

The melanin biosynthesis inhibitory effect of the B16F1 melanin forming cells was confirmed by the following method.

B16F1 melanin forming cells were divided into 6 well plates at a concentration of 2 × 10 ⁵ per well, and the cells were attached, and the cells thus attached were treated with the sample at a concentration that does not cause toxicity and incubated for 72 hours. The cultured cells were detached with trypsin-EDTA and the cells were counted and then centrifuged to recover the cells. Quantification of recovered melanin was performed by slightly modifying the method of Lotan: 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 then measured the absorbance of melanin at a wavelength of 405 nm with a microplate reader, and then quantitated melanin. The melanin production inhibition rate (%) of the sample 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 4.

IC ₅₀ values in Table 4 refer to concentrations of substances that inhibit melanin production by 50%. At this time, the hydroquinone and arbutin known to have melanin production inhibitory effect were used.

[Equation 5]

Melanin production inhibition rate (%) = [(A-B) / A] × 100

In this case, A is the melanin amount of the well to which the sample is not added, and B is the melanin amount of the well to which the sample is added.

division IC ₅₀ 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound 0.00024% Hydroquinone 0.001% Arbutin 0.02%

As can be seen in Table 4, the 3 ', 4', 5, 6, 7-penta hydroxyflavanone compound of the present invention significantly inhibited the production of melanin in B16F1 melanin forming cells, in particular Arbutin used as a positive control group It inhibited melanin production more than hydroquinone.

Experimental Example  6: Effect of Inhibiting Inflammatory Cytokine Expression by Ultraviolet Irradiation

The inhibitory effect of inflammatory cytokine expression expressed by ultraviolet irradiation of 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound prepared according to Preparation Example 1 was evaluated by the following method.

Fibroblasts isolated from human epidermal tissue (Fibroblast) were placed in a 24-well test plate, each 5 × 10 ^4, and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. . These fibroblasts were irradiated with UV 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki Co., Japan), after which PBS was removed and FBS was not added to the cell culture medium (DMEM). Medium) 350 μl was added. Each well to which the cell culture medium was added was treated with the 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound sample and incubated for 5 hours. 150 µl of the supernatant of each cultured well was taken to quantify IL-1α to determine the inhibitory effect of inflammatory cytokine expression of the sample. At this time, the amount of IL-1α was quantified using an enzyme immunoassay (Enzyme-linked Immunosorbent Assay).

Inhibition rate of the inflammatory cytokine (IL-1α) was calculated by the following Equation 6, the results are shown in Table 5.

&Quot; (6) "

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St "-Bo") / (Bt "-Bo")] × 100

In this case, Bo " is the amount of IL-1α generated in the wells not irradiated with UV light and the sample is not treated, and Bt " Is the amount of IL-1α production of the wells that were irradiated and the samples were treated.

Treatment concentration (ppm) Inhibitory rate of inflammatory cytokine expression (%) One 12.12 5 26.84 10 52.45 50 60.71 100 79.94

As can be seen in Table 5, the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound of the present invention inhibited the production of IL-1α, an inflammatory cytokine by ultraviolet light.

Manufacturing example  2 : Example  1 and Comparative example  1 ~ 2 Cosmetics  Composition manufacturing

In order to measure the effect on the cosmetic composition comprising the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound prepared according to Preparation Example 1, the 3 ', 4', 5, Cosmetic composition comprising a 6,7-pentahydroxyflavanone compound (Example 1), glycerin, 1,3- as a moisturizer instead of the 3 ', 4', 5,6,7-pentahydroxyflavanone compound A cosmetic composition including butylene glycol and sorbitol (Comparative Example 1) and a cosmetic composition not including all thereof (Comparative Example 2) were prepared in an amount as shown in Table 6 below. In this case, the content unit in Table 6 is weight%.

ingredient Example 1 Comparative Example 1 Comparative Example 2 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound 5.0 - - 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 of the cosmetic composition prepared in Preparation Example 2 (Example 1 and Comparative Examples 1 and 2) was measured as follows.

Sixteen healthy mice were divided into four groups and four animals were tested per sample. First, clean both ears of rats with ethanol, apply 4 μl of each of the cosmetic composition samples, and after 1 hour, apply 20 μl of acetone to the left ear as an experimental group and arachidone as a control to the right ear. I applied a mountain. After 1 hour, the thickness of both ears was repeatedly measured using a micrometer, and the average value thereof was obtained. The anti-inflammatory effect was calculated according to Equation 7 below, and the results are shown in Table 7.

[Equation 7]

Anti-inflammatory effect (%) = (B-A) / B × 100

At this time, A was the swelling thickness of the ear in the experimental group, B was the swelling thickness of the ear in the control group.

division Swelling Anti-inflammatory effect (%) Increased thickness Difference from control group (B-A) Control (purified water) 2.86 (B) - - Comparative Example 1 2.85 (A) 0.01 0.35 Comparative Example 2 2.71 (A) 0.15 5.24 Example 1 1.70 (A) 1.16 40.56

As can be seen in Table 7, the cosmetic composition of Example 1 containing the 3 ', 4', 5, 6, 7-pentahydroxyflavanone compound is significantly superior to the cosmetic composition of Comparative Examples 1 and 2 Indicated.

Experimental Example  8: skin irritation test

Skin irritation test of the cosmetic composition prepared in Preparation Example 2 (Example 1 and Comparative Examples 1 and 2) was measured by the following method.

Samples were prepared by adding lactic acid 0.3% as an irritant to the cosmetic compositions according to Examples 1 and Comparative Examples 1 and 2 to pH 5.5, and then, 5 × 20 The 0.3% lactic acid solution alone and 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.

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

Criteria

-: No erythema or unusual symptoms

+-: Slightly redder than the surroundings

+: Significantly redder than the surroundings

++: reddens and swells more heavily than the surroundings

[Equation 8]

Stimulus rate (%) = [(+-) number × 1 + (+) number × 2 + (++) number × 3] / [(total number) × 3] × 100

sample Judgment result Stimulation rate (%) ++ + + - - Example 1 + lactic acid 0.3% - 3 7 20 14.4 Comparative Example 1 + lactic acid 0.3% 3 15 11 One 56.66 Comparative Example 2 + 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 8, the cosmetic composition of Example 1 of the present invention in the stimulation relaxation effect experiments by lactic acid has a reduced stimulation rate compared to the control.

Experimental Example  9: moisturizing effect

Moisturizing effect on the cosmetic composition prepared in Preparation Example 2 (Example 1 and Comparative Examples 1 and 2) was measured by the following method.

After surveying 100 healthy adult men and women who feel dry skin through the survey randomly divided into three groups (A, B, C) of 20 people each, the cosmetic composition of Example 1 and Comparative Examples 1 and 2, respectively, 1 day each Apply twice to face for 8 weeks.

The 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 9 below.

sample Before use 2 weeks after use 4 weeks after use Example 1 22.3 39.9 47.9 Comparative Example 1 24.4 32.9 34.7 Comparative Example 2 22.7 30.2 31.3

As can be seen in Table 9, the cosmetic of Example 1 of the present invention was more excellent moisturizing effect than cosmetics containing a general moisturizer (Comparative Examples 1, 2).

Manufacturing example  3: Example  2-5 and Comparative example  3-4 Composition

To the cosmetic composition containing the 3 ', 4', 5, 6, 7-pentahydroxy flavanone compound of Example 1, as shown in Table 10 (Example 2-5), the 3 ', 4', 5 Cosmetics containing no, 6,7-pentahydroxyflavanone compound (Comparative Example 3) and methylparaben as an antiseptic instead of the 3 ', 4', 5,6,7-pentahydroxyflavanone compound. Cosmetics containing Dazolidinylurethane (Comparative Example 4) were prepared, respectively. At this time, the content unit of each composition component in Table 10 was weight%.

ingredient Example Comparative example 2 3 4 5 3 4 Purified water To 100 To 100 To 100 To 100 To 100 To 100 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Butylene Glycol 2.0 2.0 2.0 2.0 2.0 2.0 Propylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 Wax 4.0 4.0 4.0 4.0 4.0 4.0 Macadamia Nut Oil 5.0 5.0 5.0 5.0 5.0 5.0 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 Spices a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount 3 ′, 4 ′, 5,6,7-pentahydroxyflavanone compound 0.1 0.5 1.0 2.0 - - Methylparaben - - - - - 0.2 Imidazolidinylurea - - - - - 0.2

Experimental Example  10: antimicrobial activity test

Antibacterial effect on the bacteria and fungi of the cosmetic composition prepared according to Preparation Example 3 was measured by the following method.

First, in the case of bacteria, 20-30 g of the cosmetic composition of Example 2-5, the cosmetic composition of Comparative Example 3-4 Escherichia coli ( Escherichia 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) was mixed to add 10 ⁷ cfu / g per sample, and then cultured in a 25 ° C. incubator at 7 days intervals to observe the presence of odor and mycelia and spores on the sample surface.

The results are shown in Table 11 below.

division Bacteria (cfu / g) mold Initial number of bacteria Day 1 Day 7 Day 14 Day 21 Day 28 Example 2 1 × 107 41000 500 250 <250 <250 - Example 3 1 × 107 45000 300 200 <100 <100 - Example 4 1 × 107 44000 150 <100 <100 <100 - Example 5 1 × 107 45000 <100 <100 <100 <100 - Comparative Example 3 1 × 107 54000000 30000000 > 1 × 108 > 1 × 108 > 1 × 108 +++ Comparative Example 4 1 × 107 40000 300 <100 <100 <100 -

-: No spawning and mycelium spawning for 8 weeks and good

+: Molding on the wall or lid within 4 weeks

++: mending within 4 weeks and mold on part of the surface

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

As can be seen in Table 11, the cosmetic composition of Comparative Example 3, which did not use a preservative, the bad smell and mold on the entire surface within 4 weeks, after one day, the bacterial bacteria significantly increased. However, the cosmetic composition of Example 2-5 including the 3 ', 4', 5, 6, 7- pentahydroxyflavanone compound of the present invention showed a concentration-dependent antimicrobial effect against bacteria and fungi, After 28 days, the preservatives were similar to or better than the existing preservatives (Comparative Example 4) when used in the 2.0% content. It showed much better antiseptic power than the preservatives.

Experimental Example  11: skin wrinkle improvement

A clinical experiment was conducted on the skin wrinkle improvement effect of the cosmetic composition prepared according to Example 1. These clinical trials were evaluated through the actual use test of each cosmetic as follows.

Thirty test subjects (women aged 20 to 35 years) were applied to the cosmetic composition of Example 1 on the right side of the face, and the cosmetic composition of Comparative Example 1 on the left side of the face, twice a day for 2 consecutive months.

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 12 below.

sample Wrinkle improvement effect Effective rate (%) Great slightly none Example 1 26 3 One 96.6 Comparative Example 1 4 6 20 33.3

As can be seen in Table 12, 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 (10)

delete delete delete delete delete delete delete Preparing a compound 1 by reacting 1,3,5,6-pentahydroxybenzene and caffeic acid as in the following scheme;
Cyclizing the compound 1 to produce the following compound 2; And
Method for preparing a 3 ', 4', 5,6,7-pentahydroxyflavanone compound comprising the step of reducing compound 2 to produce compound 3 below:
[Scheme]

9. The method of claim 8,
The process for preparing the compound 2 is a method of producing a 3 ', 4', 5, 6, 7-pentahydroxy flavanone compound, characterized in that is carried out by heating after adding iodine (I ₂ ). 9. The method of claim 8,
Process for preparing the compound 3 is a method of producing a 3 ', 4', 5, 6, 7-pentahydroxy flavanone compound, characterized in that is carried out using palladium and hydrogen.

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