KR20190092098A - Cosmetic composition comprising extract, fraction or compound of Ligustri Fructuse - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of fruits of Ligustrum japonicum, a fraction thereof or a compound of ligustroside isolated therefrom as an active ingredient. The present invention can effectively inhibit melanin synthesis.

Description

Cosmetic composition comprising extract, fraction or compound separated therefrom as an active ingredient {Cosmetic composition comprising extract, fraction or compound of Ligustri Fructuse}

The present invention relates to a cosmetic composition for skin whitening containing the extract, fraction or Ligustroside (Ligustroside) compound isolated therefrom as an active ingredient.

Human skin color is largely determined by the amount of melanin, hemoglobin, carotene, and the like, of which the quinone complex called melanin, which is synthesized by melanin-forming cells in the basal cell layer of the epidermis, plays the most important role. Melanin can be divided into eumelanin and pheomelanin. Eumelanin is a granular pigment that is darker in color, darker brownish in color, larger in size, and more common in Asian or black skin. Pheomelanin is a spray-colored pigment that is light reddish brown (yellow) in color, small in size, and white in skin. Melanin is produced by enzymatic and non-enzymatic oxidation of tyrosine in melanocytes in the basal layer of the epidermis. In the melanin production mechanism, tyrosine in the cell is used as a substrate, and an enzyme called tyrosinase generates dopaquinone, which is a copolymer of melanin through autooxidation and enzymatic reaction from dopaquinone.

Melanin determines human skin color and protects skin from external stress. However, if melanin is excessively produced, pigmentation is deposited on the skin to form blemishes, freckles, spots, blotch, etc. Therefore, inhibition of the synthesis of melanin in skin whitening is a major problem.

As the cause and mechanism of melanogenesis have been revealed, it is possible to reduce the production of melanin by incorporating substances having an inhibitory effect of tyrosinase, an enzyme involved in melanogenesis, into cosmetics or inhibiting some reactions during melanogenesis. The method is generally used. Representative materials used for this purpose include plant extracts such as ascorbic acid, kojic acid, hydroquinone and lettuce extract, licorice extract. However, kojic acid has excellent tyrosinase activity inhibitory effect, while there is a problem in stability, such as lowering the titer due to discoloration and change over time when formulated in cosmetics, there is a limit on the use of large skin irritation. Ascorbic acid has a low inhibitory effect on tyrosinase activity and low stability of the molecule itself, which is not suitable as an inhibitor of melanin production. Hydroquinone has a high irritation to the skin, and its use is limited in cosmetic formulations due to safety issues. Therefore, there is a need for the development of natural substances that suppress melanin synthesis while reducing side effects.

1. Republic of Korea Patent No. 10-1627237

One object of the present invention is Ligustri Fructuse ) It is to provide a cosmetic composition for skin whitening comprising as an active ingredient extract, fraction or Ligustroside (Ligustroside) compound isolated therefrom.

In order to achieve the above object, one aspect of the present invention provides a cosmetic composition for skin whitening comprising the extract, fraction or ligustroside compounds separated therefrom as an active ingredient.

Ligustri Fructuse is a dry medicinal herb from Ligustrum lucidum Aiton, a member of the ash family. It has been used to treat symptoms and the like.

According to one embodiment of the invention, the journey room extract may be prepared according to conventional methods known in the art. Specifically, the grinding chamber may be pulverized, and then, a conventional solvent may be added, and the filtering chamber extract may be prepared under the conditions of temperature and pressure generally used.

The solvent may be selected from the group consisting of distilled water, hexane, alcohol having 1 to 4 carbon atoms, ethyl acetate, chloroform, diethyl ether, dichloromethane, acetone, and mixtures thereof, and preferably alcohol having 1 to 4 carbon atoms. .

Meanwhile, the journey extract includes not only the extract by the solvent extraction method described above, but also an extract that has undergone a conventional purification process. For example, extracts obtained through various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity, or affinity). It may also be included in the journey room extract of the present invention. In addition, the journey room extract may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying.

In the case of further fractionating the journey extract, an organic solvent selected from the group consisting of hexane, chloroform, ethyl acetate, dichloromethane and butanol may be used. For example, journey 70% ethanol extract can be sequentially partitioned into hexane, dichloromethane, ethyl acetate and butanol.

According to one embodiment of the present invention, the ligustroside compound has the structure of Formula 1 below.

According to one embodiment of the invention, the ligustroside compound may be used that is separated from the journey room or synthesized by chemical methods. When using a traveling room, the traveling room 70% ethanol extract may be sequentially partitioned into hexane, dichloromethane, ethyl acetate and butanol, and the butanol fraction may be separated by high performance liquid chromatography to obtain a ligustroside compound.

According to one embodiment of the present invention, the extract, fraction or ligustroside compound may be included in 0.01 to 10% by weight relative to the total weight of the cosmetic composition. When the extract, fraction or ligustroside compound is less than 0.01% by weight, the whitening effect of the cosmetic composition may not appear, and when it exceeds 10% by weight, the degree of increase in the whitening effect is insignificant compared to the increase in content.

The cosmetic composition of the present invention may further include components conventionally used in the cosmetic composition in addition to the above, including the extract, fraction or ligustroside as an active ingredient. Conventional adjuvants such as, for example, stabilizers, solubilizers, vitamins, pigments and flavorings, and carriers.

On the other hand, the cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants- It may be formulated with containing cleansing oil, powder foundation, emulsion foundation, wax foundation, spray, and the like, but is not necessarily limited thereto. More specifically, flexible lotion, gel, water soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water emulsion, water-in-oil emulsion, cleansing foam, cleansing water It can be prepared in the form of a pack, body oil, body lotion, oil-in-water makeup base, water-in-oil makeup base and foundation.

In case the formulation of the cosmetic composition of the present invention is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

In addition, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component when the formulation of the cosmetic composition of the present invention is a powder or a spray, and in particular, in the case of a spray, additionally chlorofluoro Propellants such as rohydrocarbon, propane / butane or dimethyl ether.

In addition, when the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate. Fatty acid esters of propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan may be used.

In addition, when the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester are used as carrier components. Suspending agents, microcrystalline cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

In addition, when the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarco Cinates, 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.

Journey extract, fraction or Ligustroside (Ligustroside) compound of the present invention can be effectively used as a cosmetic composition for skin whitening can effectively inhibit melanin synthesis.

Figure 1 shows the results of confirming the cytotoxicity of the Journey extract according to the extraction solvent.
Figure 2 shows the results confirmed the cytotoxicity of the ethanol extract 70% ethanol extract according to the treatment concentration.
Figure 3 shows the results confirming the melanin synthesis inhibitory level of the Journey extract according to the extraction solvent.
Figure 4 shows the results confirming the melanin synthesis inhibitory level of the ethanol extract 70% ethanol extract according to the treatment concentration.
Figure 5 schematically shows the process of separating the individual fractions from the journey room 70% ethanol extract.
Figure 6 shows the results of confirming the cytotoxicity of the fractions isolated from the ethanol extract of Journey room.
Figure 7 shows the results confirming the melanin synthesis inhibitory level of the fractions isolated from the ethanol extract of journey room.
Figure 8 shows the results of high performance liquid chromatography analysis of the butanol fraction separated from the 70% ethanol extract of the journey room.
Figure 9 shows the results confirming the cytotoxicity of a single peak (peak) separated from the Joules butanol fraction.
Figure 10 shows the results confirming the melanin synthesis inhibition level of a single peak separated from the Journey Butanol fraction.

Hereinafter, one or more embodiments will be described in more detail with reference to Examples. However, these examples are provided to illustrate one or more embodiments illustratively and the scope of the present invention is not limited to these examples.

Example  One: Journey Room  extract, Fraction  Produce

Water, 70% ethanol, hexane or 70% methanol was added to the journey room and extracted at 25 ° C. for 20 hours, and centrifuged at 15 ° C. (8,000 rpm, 20 minutes) to obtain respective extracts. Thereafter, each extract was lyophilized to prepare an extract powder.

Example  2: Journey Room  Confirmation of Whitening Activity of Extracts

2-1. Cytotoxicity Check

B16F10 cells, which are mouse melanocytes (melanocytes), were treated with 1 mg / ml concentration of the tremor extract prepared in Example 1 and incubated for 24 hours. After 24 hours, MTT [3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyl-tetra-zolium bromide] (Sigma Aldrich, USA) was treated at a concentration of 2 mg / ml for an additional 4 hours. Incubation was performed to dissolve formazan (DM) formed by adding DMSO (dimethylsulfoxide). Absorbance was measured at 580 nm with a Microplate reader (Molecular devices, USA) to calculate cell viability according to Equation 1 below.

As a result of confirming the cell survival rate, as shown in Fig. 1, the water extract of the fermentation chamber and the 70% ethanol extract of the fermentation chamber showed 98.7% and 91.6% cell viability, respectively, showing little cytotoxicity. On the other hand, hexane and ethanol extract 70% methanol extract showed a high cytotoxicity of 70% and 46%, respectively.

Based on the results of the experiment, the cytotoxicity of the ethanol extract 70% according to the treatment concentration was confirmed. As a result, as shown in Figure 2, even if the treatment concentration (0.25, 0.5, 1.0 and 2.0 ㎎ / ㎖) of the 70% ethanol extract of the chamber was increased, it was confirmed that there is no significant change in cell viability.

2-2. Whitening activity check

The whitening effect of the Journey extract prepared in Example 1 was tested as follows.

B16F10 cells were dispensed at ⁴ × 10 ⁴ cells / dish concentration in a 60 mm dish and then incubated for 24 hours in a 37 ° C. incubator containing 5% CO ₂ . The medium was discarded, and diluting the route extract in fresh medium was treated with B16F10 cells and further incubated for 72 hours. Then, the cells were washed with PBS (phosphte buffered saline), 1 ml of PBS was put in and scraped with a scraper (scrapper) to recover the e-tube. Cell pellets were obtained by centrifugation (7,000 rpm and 5 minutes 30 seconds) at 4 ° C., and 1 N NaOH and 10% DMSO were added to the pellets, and then left in a 60 ° C. constant temperature bath for about 30 minutes. The supernatant was placed in a 96-well plate, the absorbance was measured at 405 nm, and the melanin synthesis inhibition rate of the tremor extract was determined according to Equation 2 below.

As a result, as shown in FIG. 3, the melanin activity (synthesis amount) was reduced when the 70% ethanol extract was treated, and the melanin synthesis was inhibited by about 36%. On the other hand, 70% methanol extract inhibited melanin synthesis by about 45%, but considering the cell viability, it is determined that the melanin synthesis inhibitory effect is caused by cell death.

Since the melanin synthesis inhibitory effect of the ethanol extract 70% ethanol was the best, the melanin synthesis inhibitory activity according to the treatment concentration was confirmed.

As a result, as shown in Figure 4 it was confirmed that as the treatment concentration increases melanin synthesis inhibitory effect also increased, and when treated at 1.0 mg / ㎖ concentration melanin synthesis is inhibited by about 40%, 2.0 mg / ㎖ concentration The treatment was found to be about 60% inhibition.

Example  3: Journey Room  Isolated from 70% Ethanol Extract Fraction  Whitening activity check

3-1. Fraction  Produce

Fractions were prepared from journey room 70% ethanol extract using the procedure shown in FIG. 5.

1,500 ml of hexane was added to 70% ethanol extract (0.29% recovery, 0.431 g) in the journey room, and then centrifuged to separate the hexane fraction (15.2% recovery and 22.8 g) and the primary water fraction. 1,400 ml of dichloromethane was added to the primary water fraction, followed by centrifugation to further separate the dichloromethane fraction (0.19% recovery rate, 0.285 g) and the secondary water fraction. 1,385 ml of ethyl acetate was added to the secondary water fraction, followed by centrifugation to separate the ethyl acetate fraction (0.16% recovery rate, 0.236 g) and the tertiary water fraction. After the addition of 1,300 ml of butanol to the third water fraction, the mixture was centrifuged to separate the butanol fraction (6.22% recovery, 9.337 g) and the fourth water fraction (7.13% recovery, 10.7 g).

3-2. Fraction  Cytotoxicity Check

The cytotoxicity of each fraction isolated in 3-1 was confirmed using the same method as in Example 2-1.

As a result, as shown in FIG. 6, the hexane fraction and the dichloromethane fraction showed strong cytotoxicity at all concentrations. On the other hand, the ethyl acetate fraction and butanol fraction showed a cell viability of 80% or more when treated with 0.1 or 0.2 mg / ml concentration.

3-3. Fraction  Whitening activity check

The whitening activity of each fraction separated in 3-1 was confirmed using the same method as in Example 2-2.

As shown in FIG. 7, ethyl acetate fraction inhibited melanin synthesis by about 27% or 42% at 0.1 or 0.2 mg / ml treatment concentration, respectively. In addition, the butanol fraction at the same treatment concentration was found to inhibit melanin synthesis to about 33% or 20% level.

Example  4: Journey Room  Butanol From fractions  Confirmation of Whitening Activity of Isolated Compound

4-1. Journey Room  Butanol Fraction HLPC  analysis

From the experimental results of Example 3, it was confirmed that the butanol fraction of lysate was excellent in melanin synthesis while having low cytotoxicity. The lysate butanol fraction was analyzed by HPLC (high performance liquid chromatography, high performance liquid chromatography). As a mobile phase, a mixed solution of acetonitrile (ACN) and 0.5% formic acid was used. Mobile phase gradient is 0 min: ACN 20%; 0-10 minutes: ANC 30%; 10-20 minutes: 40% ACN; 20 to 30 minutes: ACN 50%; And 30 to 40 minutes: ACN 20% conditions.

As a result of the HPLC, it was confirmed that a single peak appeared at 21.78 minutes, 26.30 minutes and 28.07 minutes of retention time (RT) as shown in the chromatogram of FIG. 8. The peaks shown at RT 10.70 min and 13.28 min are solvent peaks.

4-2. Confirm cytotoxicity of single peak

By using the same method as in Example 2-1 was confirmed the cytotoxicity of the individual peaks isolated from the Journey butanol fraction. Single peaks appearing at RT 21.78 minutes, 26.30 minutes and 28.07 minutes are described as peak 1, peak 2 and peak 3 (the same below).

As a result, as shown in FIG. 9, except for the 1.0 mg / ml treatment concentration, it was confirmed that the cell viability was maintained at 80% or more when each peak was treated with melanocytes.

4-3. Confirm whitening activity of single peak

Using the same method as in Example 2-2, the whitening activity of the individual peaks isolated from the Journey Butanol fractions was confirmed. As a result, it was confirmed that peaks 1 to 3 exhibited melanin synthesis inhibitory effects of 49%, 29% and 9% at 0.5 mg / ml treatment concentration, respectively, as shown in FIG. 10.

4-4. Compound Analysis of Peak 1

Since the highest inhibitory effect of melanin synthesis of peak 1, a nuclear magnetic resonance (NMR) analysis was performed according to methods known in the art to identify the material of peak 1. The analysis results are as follows.

¹³ C NMR (100 MHz, MeOD-d4) δ: 173.0, 168.7, 156.8, 155.2, 130.9, 130.7, 130.5, 124.9, 116.2, 109.4, 104.4, 100.9, 95.2, 78.4, 77.9, 77.9, 75.2, 75.0, 74.8 , 72.2, 71.6, 71.5, 65.0, 62.7, 51.9, 41.3, 36.4, 31.8, 13.7.

¹ H NMR (400 MHz, MeOD-d4) δ: 7.52 (s, 1H), 7.05 (d, J = 8.4 Hz, 2H), 6.68 (s, 1H), 6.09 (d, J = 8.4 Hz, 2H) , 5.92 (s, 1H), 4.80 (d, J = 7.8 Hz, 1H), 4.35-4.29 (m, 2H), 4.23-4.18 (m, 1H), 4.02-3.87 (m, 3H), 3.73-3.65 (m, 5H), 3.47 to 3.43 (m, 2H), 3.41 to 3.33 (m, 3H), 3.21 to 3.17 (m, 1H), 2.52 to 2.46 (m, 1H), 1.72 (d, J = 7.0 Hz , 1H).

Through the NMR analysis results, it was confirmed that the material of peak 1 was Ligustroside having the structure of Formula 1 below.

[Formula 1]

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (5)

Ligustri Fructuse ) A cosmetic composition for skin whitening comprising an extract, fraction or Ligustroside compound isolated therefrom as an active ingredient.
The cosmetic of claim 1, wherein the extract is extracted with a solvent selected from the group consisting of distilled water, hexane, alcohol having 1 to 4 carbon atoms, ethyl acetate, chloroform, diethyl ether, dichloromethane, acetone, and mixtures thereof. Composition.
The cosmetic composition according to claim 1, wherein the extract, fraction or ligustroside compound is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition.
The cosmetic composition according to claim 1, wherein the fraction is fractionated with a solvent selected from the group consisting of hexane, chloroform, ethyl acetate, dichloromethane and butanol.
The method of claim 1, wherein the cosmetic composition is a flexible lotion, gel, water-soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water emulsion, water-in-oil Cosmetic composition is a formulation selected from the group consisting of emulsion, cleansing foam, cleansing water, pack, body oil, body lotion, oil-in-water makeup base, water-in-oil makeup base and foundation.

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