KR101264012B1 - Flavanone compound and Method for manufacturing the same, Cosmetic composition produced therefrom - Google Patents

Abstract

3 ', 5', 5,7-tetrahydroxy-6-methoxyflavanone (3 ', 5'), which has anti-inflammatory and antibacterial properties, can prevent skin aging or wrinkles and has a skin whitening effect. , 5,7-tetrahydroxy-6-methoxyflavanone), 3 ', 5', 5,6-tetrahydroxy-7-O-β-D-glucoflavflavone (3 ', 5', 5,6-tetrahydroxy- 7-O-β-D-glucoseflavanone) and 3 ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β-D-glucoflavflavone (3 ′, 5 ′, 5-trihydroxy- It relates to a flavanone compound comprising at least one of 6-methoxy-7-O-β-D-glucoseflavanone), a preparation method thereof, a cosmetic composition and a preparation method thereof.

Description

Flavanone compound and Method for manufacturing the same, Cosmetic composition produced therefrom

The present invention relates to a flavanone compound and a method for preparing the same, and a cosmetic composition prepared therefrom, and more particularly, to a flavanone compound and a method for producing the flavanone compound, which have anti-inflammatory and antibacterial properties, prevent skin aging and enhance skin whitening. It relates to a cosmetic composition prepared therefrom.

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 can 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, it has an anti-inflammatory and antibacterial performance, can prevent skin aging or wrinkles, and also provides a skin whitening effect and a method for producing the same, a cosmetic composition prepared therefrom The purpose.

As one aspect for achieving the above object, the present invention provides a flavanone compound characterized by the following formula.

[Chemical Formula]

In the formula, R ₁ is hydrogen (H) or glucose (Glucose), R ₂ is hydrogen (H) or methyl (CH ₃ ).

As another aspect, the present invention provides a cosmetic composition comprising the flavanone compound as an active ingredient.

In still another aspect, the present invention provides a method for preparing 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone (3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone), 3 Method for preparing ´, 5´, 5,6-tetrahydroxy-7-O-β-D-glucose flavanone (3´, 5´, 5,6-tetrahydroxy-7-O-β-D-glucoseflavanone) And 3 ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β-D-glucoflavolanone (3 ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β- D-glucoseflavanone) provides a method for producing a flavanone compound comprising at least any one of the production method.

The present invention has the following effects.

First, the flavanone compound according to the present invention and the cosmetic composition comprising the same as an active ingredient can prevent skin aging or wrinkles by inhibiting the production of MMP-1 and enhancing the synthesis of collagen.

Second, the flavanone 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 flavanone compound and the cosmetic composition comprising the same as an active ingredient has an anti-inflammatory effect and antibacterial effect against mold and bacteria by inhibiting inflammatory cytokine expression by ultraviolet irradiation.

Hereinafter, the present invention will be described in detail.

First, the flavanone compound of the present invention will be described in detail.

The flavanone compound of the present invention is represented by the following formula.

[Chemical Formula]

In the formula, R ₁ is hydrogen (H) or glucose (Glucose), R ₂ is hydrogen (H) or methyl (CH ₃ ).

In the above formula, when R ₁ is H and R ₂ is CH ₃ , it is a ₃ ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone compound.

In the above formula, when R ₁ is Glucose and R ₂ is H, it is a ₃ ′, 5 ′, 5,6-tetrahydroxy-7-O-β-D-glucoflavolanone compound.

In the above formula, when R ₁ is Glucose and R ₂ is CH ₃ , it is a ₃ ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β-D-glucose flavanone compound.

The flavanone compound of the present invention can be used for anti-aging or anti-wrinkle use.

Specifically, the flavanone compound of the present invention can be used for preventing skin aging according to the antioxidant effect as can be seen in Experimental Examples 1 and 2 to be described later, MMP as can be seen in Experimental Examples 3 and 4 to be described later It can be used for skin wrinkle improvement as it shows the effect of inhibiting -1 production and enhancing collagen synthesis.

That is, the flavanone compound, as shown in Experimental Examples 1 and 2, can prevent skin aging by preventing oxidation of skin cells and tissues by scavenging free radicals or free radicals. In addition, the flavanone compound, as shown in Experimental Example 3, by inhibiting the production of collagen by inhibiting the production of MMP-1 can prevent the decrease in skin elasticity and the formation of skin wrinkles. In addition, the flavanone compound, as shown in Experimental Example 4, by preventing collagen synthesis can be reduced skin elasticity and skin wrinkles.

The flavanone compounds of the present invention can be used for skin whitening applications.

Specifically, the flavanone compound, as can be seen in Experimental Example 5 to be described later, may be usefully used for skin whitening use by inhibiting melanin production.

That is, the flavanone compound, as shown in Experimental Example 5, can prevent skin discoloration by inhibiting the production of melanin pigment which can cause pigmentation.

The flavanone compounds of the present invention can be used for anti-inflammatory applications.

Specifically, the flavanone compound, as shown in Experimental Example 6, because it can exhibit an anti-inflammatory effect by inhibiting the inflammatory cytokine expression by ultraviolet irradiation can be usefully used for anti-inflammatory applications.

The flavanone compounds of the present invention can be used for antibacterial purposes.

Specifically, the flavanone compound, as shown in Experiment 10, as the cosmetic containing the flavanone compound as an active ingredient exhibits an antibacterial effect, it can be seen that there is an indirect antibacterial effect therefrom.

Next, the manufacturing method of the flavanone compound of this invention is demonstrated in detail.

The flavanone compound is a method of preparing 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone (3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone), 3 ′, 5 ´, 5,6-tetrahydroxy-7-O-β-D-glucoseflavanone (3 ′, 5 ′, 5,6-tetrahydroxy-7-O-β-D-glucoseflavanone) and 3 ′ 3 ', 5', 5-trihydroxy-6-methoxy-7-O-β-D-glucoseflavanone (5 ', 5-trihydroxy-6-methoxy-7-O-β-D-glucoseflavanone) It is prepared, including at least any one of the manufacturing method of).

The 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone can be prepared as in Scheme 1 below.

First, the following compound 1 is prepared by reacting 1,3,5-trihydroxy-6-methoxy benzene with caffeic acid. In this case, the preparation of Compound 1 may be carried out by the reaction at high temperature after the addition of the BF ₃ Et ₂ O catalyst.

Subsequently, the compound 1 is cyclized to prepare the following compound 2. In this case, the compound 2 may be obtained by using a DMSO solvent and cyclizing the compound 1 at a high temperature after adding iodine as a catalyst.

Subsequently, the compound 2 is reduced to prepare compound 3, that is, 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone. In this case, the compound 3 may be obtained by reacting the compound 2 with hydrogen using a palladium catalyst. Meanwhile, 3 ′, 5 ′, 5,6,7-pentahydroxyflavanone, which is Compound 4, is produced as a by-product of the preparation process of Compound 3, and Compound 4, which is a byproduct, is obtained through a conventional separation process. Can be separated from.

[Reaction Scheme 1]

The 3 ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β-D-glucose flavanone can be prepared as in Scheme 2 below.

First, compound 3 is prepared as described above.

Subsequently, the compound 3 is reacted with glucose to prepare compound 6, i.e., 3 ', 5', 5-trihydroxy-6-methoxy-7-O-β-D-glucosflavanone. In this case, Compound 6 may be prepared by adding glucose to Compound 3 using a Grignard reagent in a THF solvent.

[Reaction Scheme 2]

The 3 ′, 5 ′, 5,6-tetrahydroxy-7-O-β-D-glucose flavanone can be prepared as in Scheme 3 below.

First, compound 2 is prepared as described above.

Subsequently, the compound 2 is reduced to prepare compound 4, that is, 3 ′, 5 ′, 5,6,7-pentahydroxyflavanone. In this case, Compound 4 may be obtained by reacting Compound 2 with hydrogen using a palladium catalyst. Meanwhile, compound 3, 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone, is produced as a by-product of the compound 4 preparation process. It can be separated from compound 4.

Subsequently, the compound 4 is reacted with glucose to prepare compound 5, that is, 3 ′, 5 ′, 5,6-tetrahydroxy-7-O-β-D-glucoflavolanone. In this case, Compound 5 may be prepared by adding glucose to Compound 4 using a Grignard reagent in a THF solvent.

Scheme 3

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

The cosmetic composition includes the above-described flavanone compound as an active ingredient.

The flavanone compound may be included in an amount of 0.000001 to 30.0% by weight based on the total cosmetic composition, preferably 0.0001 to 10.0% by weight, more preferably 0.001 to 5.0% by weight. When the content of the flavanone compound is less than 0.000001% by weight, the above-described efficacy may be insignificantly expressed, and when the content of the flavanon compound exceeds 30.0% by weight, the efficacy is not significantly improved. Can fall.

The cosmetic composition according to the present invention contains components conventionally used in cosmetic compositions in addition to the flavanone compound as an active ingredient, for example, conventional auxiliaries and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. It includes.

The cosmetic composition has the effect of skin aging and wrinkle improvement, including the flavanone compound described above, has a skin whitening effect, 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 has an effect of alleviating skin irritation, as can be seen in Experimental Example 8 described later, and also has a moisturizing effect as can be seen in Experimental Example 9 described later.

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  One : Flavanon  Compound manufacturing

1) 3´, 5´, 5,7- Tetrahydroxy -6- Methoxyflavanone  Produce

As in Scheme 1, 1,3,5-trihydroxy-6-methoxy benzene and a caffeic acid derivative are reacted to obtain Compound 1, and then iodine is added to Compound 1 as a solvent, followed by heating to form a ring. Compound 2 was obtained by addition reaction. Thereafter, palladium and hydrogen were added to the obtained compound 2 to reduce the reaction to obtain a compound 3.

The obtained compound 3 was analyzed for structure using NMR and Mass analyzer. Compound 3 was identified as 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone as shown below.

yellow powder ESI-MS m / z : 319 [M + H] + 1H-NMR (MeOD - d 4, 400 MHz): 12.18 (1H, s 5-OH), 10.65 (1H, br, s, 7-OH ), 9.06 (1H, s 3'-OH), 9.01 (1H, s 5'-OH), 6.86 (1H, s, H-4 '), 6.73 (2H, s, H-2', H-6 '), 5.96 (1H, s, H-8), 5.34 (1H, dd, 12.8, 2.8 H-2), 3.66 (3H, s, -OCH3), 3.17 (1H, dd, J = 17.2, 12.4 Hz , H-3), 2.65 (1H, dd, J = 17.2, 12.4 Hz, H-3) 13C-NMR (MeOD - d 4 ,, 100 MHz): 79.21 (C-2), 42.80 (C-3) , 197.74 (C-4), 155.78 (C-5), 130.17 (C-6), 160.16 (C-7), 95.68 (C-8), 158.18 (C-9), 102.54 (C-10), 129.61 (C-1 '), 118.62 (C-2'), 146.38 (C-3 '), 115.00 (C-4'), 145.86 (C-5 '), 116.02 (C-6'), 60.6 ( OCH3).

2) 3´, 5´, 5- Trihydroxy -6- Methoxy -7-O-β-D- Of glucose flavanone  Produce

Compound 3 was obtained as described in the preparation of 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone, and as shown in Scheme 2, using Grignard reagent in a THF solvent at 0 ° C. Glucose was added and reacted with the compound 3, and the compound 6 was obtained.

The obtained compound 6 was analyzed for structure using NMR and Mass analyzer. The compound 6 was identified as 3 ′, 5 ′, 5-trihydroxy-6-methoxy-7-O-β-D-glucoflavanone as shown below.

yellow powder ESI-MS m / z : 463 [M + H] + 1H-NMR (DMSO - d6 , 400 MHz): 12.96 (1H, s, 5-OH), 9.03 (2H, s, 3 ', 5 '-OH), 6.87 (1H, d, H-4'), 6.75 (2H, s, H-2 ', H-6'), 6.30 (1H, s, H-8), 5.39 (1H, dd , J = 12.8, H-2), 4.98 (1H, d, J = 7.2), 3.68 (1H, s, -OCH3), 2.70 (1H, dd, J = 17.2, H-3α) 13C-NMR (DMSO - d6, 400 MHz): 79.51 (C-2), 43.02 (C-3), 198.49 (C-4), 149.95 (C-5), 130.84 (C-6), 159.30 (C-7), 95.16 (C-8), 158.68 (C-9), 103.96 (C-10), 130.29 (C-1 '), 118.77 (C-2'), 146.50 (C-3 '), 115.14 (C-4' ), 145.90 (C-5 '), 116.06 (C-6'), 100.5 (C-1``), 73.8 (C-2 ''), 77.8 (C-3 ''), 70.2 (C-4 ''), 77.3 (C-5``), 61.2 (C-6``), 60.96 (OCH3).

3) 3´, 5´, 5,6- Tetrahydroxy -7-O-β-D- Of glucose flavanone  Produce

Compound 2 was obtained as in the preparation of 3 ′, 5 ′, 5,7-tetrahydroxy-6-methoxyflavanone described above, and palladium and hydrogen were added to the obtained compound 2 as in Scheme 3 to reduce the reaction. Compound 4 was obtained, and then glucose was added and reacted to compound 4 using Grignard reagent in a solvent of THF at 0 ° C. to obtain compound 5.

Obtained compound 5 was analyzed for structure using NMR and Mass analyzer. Compound 5 was identified as 3 ′, 5 ′, 5,6-tetrahydroxy-7-O-β-D-glucoflavolanone as shown below.

yellow powder ESI-MS m / z : 489 [M + H] + 1H-NMR (DMSO - d6 , 400 MHz): 11.79 (1H, s, 5-OH), 9.04 (1H, s, 3'-OH ), 8.99 (1H, s, 5'-OH), 8.00 (1H, s 6-OH), 6.87 (1H, s, H-4 '), 6.74 (2H, s, H-2', H-6 '), 6.29 (1H, s, H-8), 5.36 (1H, dd, J = 15.6, 2.8, H-2), 5.08 (1H, d, J = 4.0, glu), 5.03 (1H, d, J = 5.2, glu), 4.86 (1H, d, J = 7.2, glu), 4.55 (1H, d, J = 5.6 Hz, glu), 3.17 (1H, dd, H-3α), 2.68 (1H, dd) 2.8, 6.8 Hz, H-3β) 13C-NMR (DMSO - d6 , 400 MHz): 79.46 (C-2), 43.43 (C-3), 198.74 (C-4), 149.95 (C-5), 128.5 (C-6), 155.39 (C-7), 94.95 (C-8), 154.15 (C-9), 104.12 (C-10), 130.29 (C-1 '), 118.74 (C-2'), 146.45 (C-3 '), 115.14 (C-4'), 145.90 (C-5 '), 116.02 (C-6'), 100.5 (C-1 ''), 73.8 (C-2 ''), 77.8 (C-3``), 70.2 (C-4 ''), 77.3 (C-5 ''), 61.2 (C-6 '').

Experimental Example  One. NBT Antioxidant effect measurement

The antioxidant effect of the flavanone compounds (Compounds 3, 5, 6) prepared according to Preparation Example 1 was measured using the NBT method.

That is, the active oxygen produced by xanthine and xanthine oxidase was measured by the NBT method, and the flavanon compounds of the present invention evaluated the scavenging rate for the active oxygen. 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 the vial bottle was added 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 vanone compound, 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) The blank of the flavanone compound 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 absorbance (Bt) in the same manner as in 1) except that distilled water is used instead of the flavanone compound 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 flavanone compound, Bt is absorbance at a wavelength of 560 nm after the enzymatic reaction of the blank test, and So is 560 before the reaction of the flavanone compound without enzyme addition. Absorbance at wavelength of nm, Bo is absorbance at wavelength of 560 nm before reaction in the absence of enzyme in blank test.

Treatment concentration (ppm) Active oxygen scavenging rate (%) Compound 3 Compound 5 Compound 6 One 37.1 29.1 29.1 5 55.1 47.4 47.4 10 76.1 60.2 60.2 25 86.5 73.2 73.2 50 92.1 90.7 91.7

As can be seen in Table 1, the flavanone compounds of the present invention increased the antioxidant power in a concentration-dependent manner. In particular, when the flavanone compound concentration was 50ppm or more, the antioxidant power of all the flavanone compounds was 90% or more.

Experimental Example  2 : DPPH Measurement of antioxidant activity

Antioxidant activity of the flavanone compounds (Compounds 3, 5, 6) 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 flavanone compounds is obtained by measuring the absorbance at a wavelength of 560 nm by comparing the degree of DPPH reduction by the flavanone compounds to decrease the absorbance of the blank.

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 flavanone compound to a 96-well plate, stir at high speed, and incubate at 25 ° C. for 10 minutes. Thereafter, the absorbance St 'is measured at a wavelength of 560 nm.

2) The blank of flavanone compounds is measured for absorbance (So ') in the same manner as in 1), except that methanol is used instead of 0.1 mM 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 flavanone compounds 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

Compound 3 Compound 5 Compound 6 Concentration (ppm) Free radical scavenging rate (%) Concentration (ppm) Free radical scavenging rate (%) Concentration (ppm) Free radical scavenging rate (%) One 34.6 10 72.3 10 84.6 2 61.2 50 92.8 50 93.5 5 91.5 100 92.7 100 91.2 10 95.8 250 95.6 250 87.3

As can be seen from Table 2, in the free radical scavenging test by DPPH, the flavanone compound increased the antioxidant power in a concentration-dependent manner. Compound 3 also showed better free radical scavenging rates at much lower concentrations than compounds 5 and 6.

Experimental Example  3: MMP -1 suppression effect

The inhibitory effect of MMP-1 production, a collagen degrading enzyme, of the flavanone compounds (Compounds 3, 5, 6) 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, the experimental group was further incubated for 48 hours in serum-free DMEM medium to which the flavanone compounds were added and serum-free DMEM medium to which no flavanone compounds were added as a control group.

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 3.

&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 (%) Compound 3 Compound 5 Compound 6 One 37.1 29.1 29.1 2.5 55.1 47.4 47.4 5 76.1 60.2 60.2 10 86.5 73.2 73.2 25 92.1 90.7 91.7

As can be seen in Table 3, all the flavanone compounds reduced the activity of MMP-1 in a concentration-dependent manner.

Experimental Example  4: collagen synthesis promoting effect

Collagen synthesis enhancing effect of the flavanone compounds (Compounds 3, 5, 6) 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 cells were further incubated for 48 hours in serum-free DMEM medium to which the flavanone compounds were added as an experimental group and serum-free DMEM medium not containing the flavanone compounds as a control group.

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 4.

&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 (%) Compound 3 Compound 5 Compound 6 One 5.31 4.34 5.78 2.5 14.31 11.31 12.64 5 31.61 34.42 39.63 10 55.78 54.04 61.31 25 71.84 76.54 82.32

As can be seen in Table 4 above, all of the flavanone compounds increased collagen production rate in a concentration-dependent manner.

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

Skin whitening effect of the flavanone compounds (Compounds 3, 5, 6) 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 flavanone compounds during artificial culture of the cells.

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

B16F1 melanin forming cells were divided into 6 well plates at a concentration of 2 × 10 ⁵ per well and attached to the cells, and the cells thus attached were treated with the flavanone compounds at a concentration which 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 flavanone compound 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, 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 amount of melanin in the well to which the flavanone compounds are not added, and B is the amount of melanin in the well to which the flavanone compounds are added.

sample IC ₅₀ Compound 3 0.0049% Compound 5 0.0043% Compound 6 0.0032% Hydroquinone 0.001% Arbutin 0.02%

As can be seen from Table 5, the flavanone compounds significantly inhibited the melanin production of B16F1 melanin forming cells, and in particular, it was more effective than arbutin used as a positive control and showed a hydroquinone-like effect.

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

The inhibitory effect of inflammatory cytokine expression expressed by UV irradiation of flavanone compounds (Compounds 3, 5, 6) 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 flavanone compounds and incubated for 5 hours. The supernatant of each cultured well was taken to quantify IL-1α to determine the inhibitory effect of inflammatory cytokine expression of the flavanone compounds. 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 6.

&Quot; (6) "

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

In this case, Bo "is the amount of IL-1α produced in the wells not irradiated with ultraviolet rays and not treated with flavanone compounds, and Bt" is the amount of IL-1α produced in the wells irradiated with ultraviolet rays and not treated with flavanone compounds. St ″ is the amount of IL-1α produced in wells treated with UV light and treated with flavanone compounds.

Treatment concentration (ppm) Inhibitory rate of inflammatory cytokine expression (%) Compound 3 Compound 5 Compound 6 One 5.4 3.2 2.9 5 10.4 11.6 9.9 10 31.1 29.4 28.3 50 61.4 62.7 59.7 100 75.6 78.3 77.7

As can be seen in Table 6, the flavanone compounds inhibited the production of IL-1α, an inflammatory cytokine by ultraviolet light.

Manufacturing example  2 : Example  1-3 and Comparative example  1-2 Cosmetics  Composition manufacturing

In order to measure the effect on the cosmetic compositions comprising the flavanone compounds prepared according to Preparation Example 1 as an active ingredient, the cosmetic compositions (Examples 1 to 3), glycerin as a moisturizer instead of the flavanone compounds , 1,3-butylene glycol and a cosmetic composition (Comparative Example 1) comprising a sorbitol and a cosmetic composition (Comparative Example 2) not containing all of them were prepared in the content shown in Table 7. In this case, the content unit in Table 7 is the weight percent.

ingredient Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Compound 3 2.0 - - - - Compound 5 - 2.0 - - - Compound 6 - - 2.0 - glycerin - - 5.0 - 1,3-butylene glycol - - - 2.5 - Sorbitol - - - 2.5 - EDTA-2Na 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 Microcrystallin 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 3.0 3.0 Trioctanoine 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 Incense, preservative Suitable amount Suitable amount 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 compositions prepared in Preparation Example 2 (Examples 1 to 3 and Comparative Example 1) 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 compositions, and after 1 hour, apply 20 μl of acetone to the left ear to the experimental group and arachidonic acid as a control to the right ear. Applied. 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 8.

[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.

sample Swelling Anti-inflammatory effect (%) Increased thickness Difference from control group (B-A) Control (purified water) 2.87 (B) - Comparative Example 1 2.84 (A) 0.03 1.04 Example 1 1.39 (A) 1.48 51.57 Example 2 1.30 (A) 1.57 54.70 Example 3 1.20 (A) 1.67 58.19

As can be seen in Table 8, the cosmetic composition of Examples 1 to 3 containing the flavanone compounds of the present invention showed an excellent anti-inflammatory effect than the cosmetic composition of Comparative Example 1.

Experimental Example  8: skin irritation test

Skin irritation test of the cosmetic compositions prepared in Preparation Example 2 (Examples 1 to 3 and Comparative Example 1) was measured by the following method.

Samples were prepared by adding 0.3% lactic acid as a stimulating agent to the cosmetic composition according to Examples 1 to 3 and Comparative Example 1 so that the pH was 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, 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

[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% - One 2 27 4.44 Example 2 + lactic acid 0.3% - 2 2 26 6.66 Example 3 + lactic acid 0.3% - 2 2 26 6.66 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, the cosmetic compositions of Examples 1 to 3 of the present invention in the stimulation relaxation effect experiments by lactic acid decreased the stimulation rate compared to the control cosmetic composition.

Experimental Example  9: moisturizing effect

Skin irritation test of the cosmetic compositions prepared in Preparation Example 2 (Examples 1 to 3 and Comparative Examples 1 and 2) was measured by the following method.

The survey divided 100 healthy adult men and women who feel dry skin into 5 groups (A, B, C, D, E) of 20 randomly. The cosmetic compositions of Examples 1, 2, and 3 were applied to the D and E groups on the face for 8 weeks, twice a day for the cosmetic compositions of Comparative Examples 1 and 2.

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 10 below.

sample Before use 2 weeks after use 4 weeks after use Example 1 23.1 39.9 47.2 Example 2 23.4 38.7 43.2 Example 3 23.2 38.8 46.5 Comparative Example 1 24.3 29.8 31.5 Comparative Example 2 22.7 31.4 31.7

As can be seen in Table 10, the cosmetic compositions of Examples 1 to 3 of the present invention was more excellent than the cosmetic compositions of Comparative Examples 1 and 2 including a general moisturizing agent.

Manufacturing example  3: Example  4-7 and Comparative example  3-4 composition preparation

As shown in Table 11, each of the flavanon compounds prepared according to Preparation Example 1 and the cosmetic compositions (Examples 4 to 7) containing all of them as an active ingredient, and do not include the flavanone compounds Instead of the cosmetics (Comparative Example 3) and the flavanone compounds, a cosmetic (Comparative Example 4) containing methylparaben and imidazolidinylurethane as preservatives was prepared, respectively. At this time, in Table 11, the content unit of each composition component was weight%.

ingredient
Example Comparative example 4 5 6 7 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 Compound 3 0.2 - - 0.1 - - Compound 5 - 0.2 - 0.1 - - Compound 6 - - 0.2 0.1 - - 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 4-7 and the cosmetic composition of Comparative Example 3-4 ( E. coli) 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 12 below.

sample Bacteria (cfu / g) mold
Initial number of bacteria Day 1 Day 7 Day 14 Day 21 Day 28 Example 4 1 x 10 ⁷ 41000 300 200 <100 <250 - Example 5 1 x 10 ⁷ 45000 300 200 <100 <100 - Example 6 1 x 10 ⁷ 44000 250 <100 <100 <100 - Example 7 1 x 10 ⁷ 45000 <100 <100 <100 <100 - Comparative Example 3 1 x 10 ⁷ 54000000 30000000 > 1 × 10 ⁸ > 1 × 10 ⁸ > 1 × 10 ⁸ +++ Comparative Example 4 1 x 10 ⁷ 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 12, the cosmetic composition of Comparative Example 3, which does not use a preservative, the bad smell and mold on the entire surface within 4 weeks, bacterial bacteria significantly increased even after one day. However, the cosmetic compositions of Examples 4-7 including flavanone compounds of the present invention showed a concentration-dependent antimicrobial effect against bacteria and fungi, and Example 7 including all flavanone compounds by 0.1% each was 28 After one day, the same or better preservative strength as Comparative Example 4 including the preservative was shown.

Experimental Example  11: skin wrinkle improvement

Clinical trials were conducted on the skin wrinkle improvement effects of the cosmetic compositions according to Examples 1 to 3. These clinical trials were evaluated through the actual use test of each cosmetic composition as follows.

Thirty test subjects (women aged 20 to 35 years) were applied to the cosmetic composition of Examples 1 to 3 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 13 below.

sample Wrinkle improvement effect Effective rate (%) Great slightly none Example 1 25 4 0 100% Example 2 25 5 0 100% Example 3 26 4 0 100% Comparative Example 1 5 4 21 30

As can be seen in Table 13, the cosmetic compositions according to Examples 1 to 3 of the present invention showed a higher wrinkle improvement effect than the cosmetic composition according to Comparative Example 1. In addition, skin irritation could not be observed in the skin of the subjects applying the cosmetic compositions according to Examples 1 to 3 on the skin.

Claims (13)

delete delete delete delete Flavanone compounds represented by the formula:
[Chemical Formula]

(In the above formula, R ₁ is hydrogen (H) or glucose (Glucose), R ₂ is hydrogen (H) or methyl (CH ₃ ))
Cosmetic composition for preventing skin aging comprising as an active ingredient. Flavanone compounds represented by the formula:
[Chemical Formula]

(In the above formula, R ₁ is hydrogen (H) or glucose (Glucose), R ₂ is hydrogen (H) or methyl (CH ₃ ))
Cosmetic composition for improving skin wrinkles containing as an active ingredient. Flavanone compounds represented by the formula:
[Chemical Formula]

(In the above formula, R ₁ is hydrogen (H) or glucose (Glucose), R ₂ is hydrogen (H) or methyl (CH ₃ ))
Cosmetic composition for skin whitening comprising as an active ingredient. delete The cosmetic composition according to any one of claims 5 to 7, wherein the flavanone compound is contained in an amount of 0.000001 to 30% by weight based on the total weight of the cosmetic. delete delete delete delete