KR101068976B1 - Manufacturing method of nelumbinis semen extract with alpha-glucosidase inhibitory effect and cosmetic composition comprising the extract - Google Patents

Abstract

The present invention relates to a method for producing a specific extract of lotus root meat, which inhibits alpha-glucosidase enzyme activity, characterized in that the extract of lotus root is extracted with an aqueous ethanol solution and fractionated into butanol. The soft extracts of the soft muscle prepared by the preparation method according to the present invention have a markedly improved effect of inhibiting alpha-glucosidase and tyrosinase activity and inhibiting melanogenesis.

Description

Manufacturing method of Nelumbinis semen extract with alpha-glucosidase inhibitory effect and cosmetic composition comprising the extract}

The present invention relates to a cosmetic composition for whitening that inhibits alpha-glucosidase enzyme activity and a manufacturing method for enhancing the whitening effect of certain herbal medicines.

In general, the skin color is determined by the content, distribution, and the like of melanin. In particular, it is associated with the number and distribution of melanosomes produced in intracellular melanocytes and released out of the cell. Melanin is widely present in animals, plants and microorganisms as an important defense means of protecting the living body from harmful ultraviolet rays in sunlight. However, hyperpigmentation of the skin may be caused by various factors such as hormonal abnormalities, genetic diseases, and ultraviolet irradiation after inflammatory reactions of the skin. The main factors are due to abnormalities in melanin synthesis and distribution.

The melanin production process is a series of oxidative polymerization reactions starting from tyrosine, a relatively stable substance. In this process, tyrosinase plays the most important role. Dopa quinone, an intermediate produced by the action of this enzyme, is unstable and is oxidatively polymerized into melanin via dopa chrome.

Recently, the screening method of whitening substances interferes with melanin biosynthesis mechanisms such as inhibitors of existing tyrosinase activity, alpha-melanocyte stimulating hormone antagonists, and inhibition of N-glycosylation. There is also a great deal of research in the field of chemicals ( The Biochemical Journal , 344 (3), 659-665, 1999).

Tyrosinase, which plays an important role in melanin biosynthesis, is a glycoprotein and is produced through glycosylation, a glycoprotein synthesis process in vivo. If there is a problem in the glycosylation process and an abnormality occurs in the sugar portion of tyrosinase, the tyrosinase does not move to the melanosomal, a place of intracellular melanin biosynthesis, or it does not show the activity of tyrosinase. Creation cannot be made. Article lycopene actual migration process are those of the enzyme alpha important for many enzymes involved - the Cedar gluconate kinase (The Journal of Biological Chemistry , 272 (25), 15796-15803, 1997). If the activity of the enzyme can be inhibited, the glycosylation process of tyrosinase is inhibited and melanin production will not be achieved, which may have a whitening effect.

Therefore, the technical problem to be achieved by the present invention is to provide a cosmetic composition for whitening exhibiting a whitening effect by inhibiting alpha-glucosidase.

Another technical problem to be achieved by the present invention is to provide a method for producing a cosmetic composition excellent in whitening effect.

In order to achieve the above technical problem, the present invention provides a cosmetic composition for skin whitening, characterized in that it contains as an active ingredient soft lotus extract, in particular, soft extract produced by a specific manufacturing method.

Lotus is the fruit of Nelumbo nucifer a Gaertner, a perennial herb belonging to the water lily family, which is a long gray oval string of dark gray or dark brown color. The color is light red on the outside, milky white or sulfur on the inside, in which the turquoise embryo is kept and the quality is solid. Remove embryos and use only meat.

The inventors of the present invention, while studying a cosmetic composition using natural products, the extract of lotus root is excellent in the inhibitory effect of alpha-glucosidase and tyrosinase activity and melanin production, and even more surprisingly, the whitening of the lotus root extract prepared by a specific manufacturing method The present invention was completed by finding that the effect is excellent.

Therefore, preferably, the present invention is to extract the butanol fraction of the extract prepared by first extracting the soft meat with an ethanol aqueous solution (preferably, 50-90 wt% ethanol aqueous solution, more preferably 70-80 wt% ethanol aqueous solution) The present invention provides a cosmetic composition for skin whitening, wherein the inhibitory effect of alpha-glucosidase and tyrosinase activity and the inhibition of melanin biosynthesis of lotus root extract prepared by this method are significantly higher than those of other extracts outstanding.

The present invention also provides a step of preparing a primary extract by extracting (S1) soft-edible extract with ethanol aqueous solution; And (S2) provides a method for producing a whitening effect for improving the whitening and the whitening effect of the whitening and extracting the whitening effect of the first extract with butanol, and preferably a method for enhancing the whitening effect of the extract , The ethanol aqueous solution of step (S1) is an ethanol aqueous solution of 50-90% by weight of ethanol, more preferably an ethanol aqueous solution of 70-80% by weight of ethanol.

More preferably, in the above method, the primary extract is decompressed and concentrated and again dissolved in an aqueous ethanol solution having an ethanol content of 5-20% by weight (more preferably, 7-13% by weight), followed by step (S2). It provides a method characterized in that it is used for fractionation, and the use of this specific method can further increase the whitening effect of the lotus root extract more than expected. Most preferably, the first extraction is carried out for 4 to 20 hours at a temperature condition of 50-95 ℃, the reduced pressure and concentration is a rotary evaporator is used.

The present invention also provides a cosmetic composition for skin whitening comprising as an active ingredient the extract of lotus root meat prepared by the production method according to the present invention.

In the cosmetic composition of the present invention, the active ingredient extract of soft lotus meat is preferably included in the amount of 0.01 to 50% by weight relative to the total weight of the cosmetic composition. If the content of the extract is less than 0.01% by weight, it is not preferable because it is not able to sufficiently achieve the desired useful effect originally, and if the content of the extract as the active ingredient exceeds 50% by weight relative to the total weight of the cosmetic, It may cause a problem, and thus there is a concern that the cosmetic appearance or the use of the cosmetic may not be sufficient cosmetic effects.

The cosmetic composition prepared by containing the soft extract of the extract is not particularly limited in the formulation, for example, cosmetics, essences, lotions, creams, packs, foundations, such as foundations, makeup bases, color tone It can be prepared in various formulations of the product.

The cosmetic composition of the present invention, in addition to the lotus extract according to the present invention in the range that does not inhibit the effect of the present invention, moisturizers, thickeners, surfactants, emulsifiers, preservatives, antioxidants, alcohols, fragrances, pH adjusters, natural extracts, etc. Of course, other chemical ingredients for imparting effects such as skin moisturizing and skin aging can be blended in an appropriate amount.

The present invention provides a cosmetic composition for skin whitening, characterized in that it contains an extract of the soft lotus meat prepared by a specific manufacturing method as an active ingredient. The present invention also provides a specific manufacturing method that can increase the whitening effect of the lotus extract.

Hereinafter, examples and the like will be described in detail to help understand the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

Example  1 to 5

After putting 50 g of lotus root meat into the extractor, 500 mL of 75% ethanol aqueous solution was added thereto, followed by repeated reflux cooling extraction twice for 4 hours. The extract filtrate was filtered through Whatman No. 2 filter paper and concentrated under reduced pressure with a rotary evaporator to obtain an ethanol extract (Example 1). After dissolving this by adding 1 L of 10% aqueous ethanol solution, the same amount of dichloromethane, ethyl acetate, butanol and water were sequentially added to obtain Examples 2 to 5 (see Table 1).

division Extraction Method Example 1 75% ethanol solution Example 2 Dichloromethane fractionation layer Example 3 Ethyl acetate fractionation layer Example 4 Butanol fractionation layer Example 5 Water fractionation layer

Test Example  1: alpha Glucosidase  Activity Inhibitory Effect Experiment

100 μL of 0.1 M phosphate buffer solution and 200 μL of any one of Examples 1 to 5, acarbose and deoxynojirimycin were added to 10 μL of an enzyme solution of alpha-glucosidase (Megazyme Co., Ltd.). It left for 37 minutes at 37 degreeC. 100 μL of the substrate 2 mM ρ-nitrophenyl-α-D-glucopyranoside was added thereto, followed by enzymatic reaction at 37 ° C. for 10 minutes, and the absorbance was measured at 405 nm. At this time, acarbose and deoxynorimycin were used as positive controls.

Using the following equation 1 to calculate the enzyme activity inhibition rate as shown in Table 2, and the experiment was performed three times each to represent the average value ± standard deviation. In the following table, IC ₅₀ is a concentration of the sample required to inhibit alpha-glucosidase activity by 50%, which is a value used in comparison. The smaller the value, the better the activity inhibition rate.

Test substance IC ₅₀ (μg / mL) Example 1 1.908 ± 0.195 Example 2 12.429 ± 1.332 Example 3 1.570 ± 0.226 Example 4 0.558 ± 0.081 Example 5 8.550 ± 1.569 Akabos 447.731 ± 18.904 Deoxynojirimycin 569.799 ± 15.605

As shown in Table 2, Example 3 in the inhibitory effect of alpha-glucosidase activity showed a slightly increased inhibitory effect compared to Example 1, and in Example 2 and 5 showed a reduced effect. On the other hand, Example 4, which is a specific extract, significantly improved the effect of inhibiting alpha-glucosidase activity. In addition, Examples 1 to 5 of the present invention was confirmed to have a superior alpha-glucosidase activity inhibitory effect compared to the positive control group acarbose and deoxynojirimycin.

Test Example  2: Experiment of Inhibitory Effect of Tyrosinase Activity

In order to measure the whitening effect of the extract of the present invention, the effect of inhibiting tyrosinase activity was measured. Examples 1 to 5 0.9 mL, 0.1 mL of 0.1M phosphate buffer (pH 6.8) and 1.0 mL of 1.5 mM L-tyrosine solution were mixed well, and warmed up at 37 ° C. for 10 minutes. After 0.1 mL of mushroom tyrosinase (1,500 units / mL) was added and reacted at 37 ° C for 10 minutes, the absorbance was measured at 475 nm.

By using Equation 2 to calculate the enzyme activity inhibition rate as shown in Table 3 below, the experiment was performed three times each to represent the average value ± standard deviation. The values shown in the table below are expressed as tyrosinase activity inhibition rates at 1 mg / mL concentrations of Examples 1-5.

Test substance Inhibition rate of tyrosinase activity (%) Example 1 2.174 ± 0.283 Example 2 7.341 ± 2.138 Example 3 13.062 ± 2.274 Example 4 32.723 ± 2.961 Example 5 7.468 ± 3.695

As shown in Table 3, Examples 2 to 5 of the present invention showed an improved inhibitory effect on tyrosinase activity compared to Example 1, in particular, Example 4 showed a significantly improved tyrosinase inhibitory effect .

Test Example  3: mouse Melanoma  Inhibitory Effects of Melanin Biosynthesis on Cells

B16F1 melanoma cells were dispensed in Dwell (Dulbecco's Modified Eagle's Medium, HyClone Lab., USA) with 10% bovine serum to 1 x 10 ⁵ cells / well in 6 well plates at 5% CO ₂ , 37 ° C. Incubated for 24 hours. After removing the culture solution and replacing Example 4 by diluting the medium to 20, 50, 100 μg / mL concentration, and added to the final 0.2 μM α-MSH (melanocyte stimulating hormone, Sigma) and further incubated for 48 hours . As a control, only the α-MSH was added without processing the sample. After incubation, the culture medium was removed, washed with PBS, 1N NaOH containing 10% DMSO was added, and intracellular melanin was dissolved in a 50 ° C thermostat. This solution was measured for absorbance at 490 nm using a microplate reader and corrected with total protein. The experiment was performed three times each, and the average value ± standard error is shown in Table 4 below, and the measured value of the control group was 100%.

Test substance Melanin biosynthesis rate (%) 20 μg / mL 50 μg / mL 100 μg / mL Control group 100 100 100 Example 3 115.245 ± 4.072 100.600 ± 2.852 70.132 ± 4.594 Example 4 82.160 ± 3.614 56.092 ± 5.354 54.291 ± 1.070 Arbutin 72.637 ± 6.837 55.584 ± 7.516 40.610 ± 3.155

As shown in Table 4, Example 4 showed an excellent melanin biosynthesis inhibitory effect.

Formulation example  1 and Comparative formulation example  One

The composition of the whitening skin containing the lotus leaf extract according to Example 4 was prepared as shown in Table 5 below. At this time, the unit of component content is weight%. On the other hand, of the components distinguished by each component number in Table 5 below, components 2 and 3 are first dispersed and dispersed in component 1 (purified water), and then components 4 to 6 are added, and components 7 to 9 are added and mixed in a manner. Prepared. As a comparative formulation example, the remainder of the component constitution except for component 9 and the preparation method thereof were produced in the same manner.

number ingredient Formulation Example 1 Comparative Formulation Example 1 One Purified water Balance Balance 2 Butylene Glycol 2.0 2.0 3 glycerin 1.0 1.0 4 Fiji-1500 0.5 0.5 5 ethanol 4.0 4.0 6 Polysorbate 80 0.2 0.2 7 incense 0.1 0.1 8 antiseptic a very small amount a very small amount 9 Lotus root extract according to Example 4 1.0 -

Formulation Example 2 and Comparative Formulation 3

The composition of the whitening cream containing the soft extract according to Example 4 was prepared as shown in Table 6 below. At this time, the unit of component content is weight%. On the other hand, among the components identified by the component numbers in Table 6 below, components 1 to 7 were first dissolved by heating at a temperature of 70 ° C., and then components 9 to 12 were dissolved and dispersed in component 13, and emulsified in those heated to 70 ° C. . Thereafter, the emulsified product was cooled to a temperature of 56 ° C., and then component 14 was added thereto, stirred, and cooled to room temperature.

number ingredient Formulation Example 3 Comparative Formulation Example 3 One Cetearyl Alcohol 1.0 1.0 2 Glyceryl Stearate / Pig-100stearate 1.0 1.0 3 Polysorbate 60 1.0 1.0 4 Sorbitan sesquioleate 0.3 0.3 5 Cetyloctanoate 6.0 6.0 6 Squalane 8.0 4.0 7 Apricot Kernel Oil 4.0 4.0 8 Dimethicone 2.0 2.0 9 glycerin 5.0 5.0 10 Magnesium aluminum silicate 0.4 0.4 11 Xanthan Gum 0.03 0.03 12 antiseptic a very small amount a very small amount 13 Purified water Balance Balance 14 Lotus root extract according to Example 4 5.0 -

Test Example  4: Skin whitening  Effect and skin irritation sensory test

In order to evaluate the skin whitening improvement effect and skin irritation of the cosmetic composition for skin whitening according to the present invention, a sensory test was conducted using the whitening cream prepared in Formulation Example 2 and Comparative Formulation Example 2. Specifically, in order to measure the whitening effect of the skin when the whitening cream of Formulation Example 1 and Comparative Formulation Example 1 were applied to the skin, 10 women over 20 years of age were used continuously twice a day for 4 weeks.

For the skin whitening effect item in the sensory test, the skin whitening effect of the whitening cream of Formulation Example 1 was relatively evaluated based on the whitening cream of Comparative Formulation Example 1. The sensory evaluation on skin irritation was itching and itching. And phenomena such as erythema. The evaluation was performed based on the five-point method of very good (5 points), excellent (4 points), moderate (3 points), bad (2 points), very bad (1 point), and the results are shown in Table 7 below. Indicated. Skin irritation in Table 7 represents the degree of no skin irritation.

number Skin irritation Skin whitening effect Formulation Example 1 Comparative Formulation Example 1 One 4 4 5 2 5 5 5 3 4 4 4 4 5 4 5 5 4 5 5 6 5 5 4 7 4 4 5 8 5 5 5 9 4 5 5 10 5 5 5 Average 4.5 4.6 4.8

As shown in Table 7, the skin irritation evaluation score for the cosmetic composition of Formulation Example 1 according to the present invention was evaluated very good as 4.5, confirming that the skin irritation degree was low as in Comparative Formulation Example 1 and thus the skin safety was excellent. Could.

In addition, the relative skin whitening effect of the cosmetic composition of Formulation Example 1 compared to Comparative Formulation Example 1 was found to be very excellent with an evaluation score of 4.8 points.

Claims (6)

A cosmetic composition for skin whitening, characterized in that it contains an extract of lotus root meat, which is a butanol fraction of an extract primarily extracted with an aqueous ethanol solution, which has an effect of inhibiting the activity of alpha-glucosidase. delete The cosmetic composition for skin whitening according to claim 1, wherein the ethanol aqueous solution is an ethanol aqueous solution having an ethanol content of 70-80 wt%. (S1) preparing a primary extract by extracting an extract of soft lotus meat with an ethanol aqueous solution; And
(S2) fractionating the primary extract with butanol
Method for producing an excellent white meat extract, characterized in that it comprises a whitening effect. The method of claim 4, wherein the aqueous ethanol solution of step (S1) is an ethanol aqueous solution having an ethanol content of 70-80% by weight. 5. The method according to claim 4, wherein the primary extract is decompressed and concentrated and dissolved again in an aqueous ethanol solution having an ethanol content of 5-20% by weight and used for fractionation in step (S2).