KR101256180B1 - Cosmetic composition comprising extract of thin film outer cover of Luffa Roemer seed for skin whitening - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening containing a thin skin coat extract of Smear cucumber seed as an active ingredient. Thin-film outer skin extracts of the scrubber seed according to the present invention significantly lower melanocyte viability than the scrubber meat extract, better inhibit the tyrosinase activity in melanocytes, and further reduce the melanin content in the melanocytes, skin whitening It can be usefully used in the cosmetic composition for.

Description

Cosmetic composition comprising extract of thin film outer cover of Luffa Roemer seed for skin whitening}

The present invention relates to a cosmetic composition for skin whitening containing a thin skin coat extract of Smear cucumber seed as an active ingredient.

The skin color of a person is determined by the concentration and distribution of melanin in the skin. The melanin pigment, which is produced in melanocytes of human skin, is a phenolic high molecular substance having a complex form of black pigment and protein and plays an important role in blocking skin damage caused by ultraviolet rays. It is reported that the action of tyrosinase in melanocytes is the most important for melanin biosynthesis, and tyrosinase is a type of amino acid tyrosine (tyrosine) and dopa (DOPA, dihydroxyphenylalanine), an intermediate of melanin polymer production. Conversion to dopaquinone plays a key role in the skin blackening process.

When such synthesis of melanin occurs excessively in the skin, it may darken the skin tone and cause blemishes and freckles. Therefore, by inhibiting the synthesis of melanin pigment in the skin, not only can brighten the skin tone to realize skin whitening, but also improve skin hyperpigmentation such as spots, freckles, etc. caused by ultraviolet rays, hormones and genetic causes. have.

Conventionally, in order to improve skin whitening or hyperpigmentation, substances having inhibitory activity against tyrosinase such as hydroquinone, ascorbic acid, kojic acid, glutathione and cysteine are used. Has been used. However, hydroquinone exhibits a predetermined whitening effect, but has a problem of limiting the amount of the compound to a very small amount due to severe skin irritation, and ascorbic acid is easily oxidized, causing problems such as discoloration / deodorization in the cosmetics containing it. There is a disadvantage that the manufacturing process of the cosmetic is complicated by the instability in the silver solution. In addition, thiol-based compounds such as glutathione and cysteine not only have a characteristic unpleasant odor, but also have problems with transdermal absorption, and glycosides and derivatives thereof have high polarity, making it difficult to use as a component of a cosmetic composition. In addition, vitamin C has a problem in that it is easily oxidized in the aqueous solution state does not have a lasting effect.

Therefore, in recent years, a lot of research on cosmetic compositions for skin whitening including natural herbal extracts.

On the other hand, Luffa is an annual plant of the gourd, which is also called a loofah, sponge gourd, aquatic plant, aquatic plant, gulf, cheonraksa (天 천) or cheonra (天 羅). The scrubber components so far identified include ash, which supplies nutrients to various cells, regulates osmotic pressure, carotene, which lowers heat and detoxifies, and proteins that produce immune and blood and bleed. In addition, it contains various ingredients such as dietary fiber, calcium, potassium, iron, magnesium and essential amino acids. In addition, the scrubber cucumber is so cold that it removes phlegm that causes a lot of heat in the body. By cooling hot blood, it promotes blood circulation, it is effective for inflammation of acne, sweat, etc., it is anti-inflammatory, it is helpful for preventing constipation and treating sinusitis, it is effective for lung diseases, hangover, summer cold, It is effective for headache, soothing effect, prevents excessive secretion of sebum, prevents clogging of pores, provides sufficient moisture to the skin, and has an excellent anti-itch effect. Antimicrobial and skin cell regeneration effects are known. In addition, scrubber seeds and leaves have a diuretic and detoxification effect, the skin and roots are known to have analgesic anti-inflammatory action. The sap coming from the cut surface of the stem of the loofah is edible and also used as a raw material for cosmetics.

As described above, the various physiological activities of the scrubber cucumber mainly use leaves, fruits, seeds, stems, and the like, and the thin film shells surrounding the seeds of the scrubber cucumber have not been studied.

Therefore, the necessity of the study of the thin film shell which wraps up the seed of the scrubber cucumber discarded is calculated | required.

The inventors of the present invention while studying the thin film shell covering the seed of the discarded scum cucumber cucumber, the thin film shell extract of the scum cucumber cucumber seed has lower melanocyte viability than the scum cucumber meat extract, and the effect of inhibiting tyrosinase activity in melanocytes is excellent. , It was confirmed that the melanin reduces the melanin content in the cells, and completed the present invention.

Therefore, the present invention is to provide a cosmetic composition for skin whitening containing a thin skin coat extract of Smear cucumber seed as an active ingredient.

The present invention provides a cosmetic composition for skin whitening containing a thin skin coat extract of Smear cucumber seed as an active ingredient.

Hereinafter, the present invention will be described in detail.

The thin skin coat extract of the scrubber seed of the present invention is extracted and used as follows. First, peel off the white area surrounding the seedlings of the scumber cucumber seed (seed bark is black with the seed shell), collect it, add it to distilled water, and heat it over a low heat of about 50 to 70 ° C for 1 to 3 hours. The thin film skin of the heated scrubber seed is taken out, drained and powdered, and then put in distilled water, treated with an ultrasonic cleaner, filtered and lyophilized to obtain a thin film skin extract of the scrubber seed in powder form.

Freeze-drying is a method of freezing a sample containing an aqueous solution or water, depressurizing it to below the water vapor pressure at that temperature, and subliming water to dry it. The freeze dryer consists of a sample drying chamber into which a sample is frozen in advance, a vacuum pump or a diffusion pump, and a condenser which condenses and removes the sublimed water vapor in between. Freeze-drying occurs at low temperatures, so it is suitable for concentration of aqueous solution because of less deterioration of the sample due to heat and good solubility of the sample after drying. In addition, lyophilization allows long-term, stable preservation of samples with unstable biological activity in aqueous or aqueous solutions.

The thin-film sheath extract of the scrubber seed according to the present invention significantly lowers melanocyte viability than the scrubber meat extract, has a better inhibitory effect on tyrosinase activity in melanocytes, and further reduces melanin content in melanocytes. Therefore, the thin skin coat extract of the scrubber seed according to the present invention can be usefully used in the cosmetic composition for skin whitening.

The cosmetic composition for skin whitening of the present invention may contain one or more known active ingredients having a skin whitening effect together with the thin skin coat extract of the scrubber seed.

The cosmetic composition for skin whitening of the present invention, in order to have a skin whitening effect, in addition to the thin film skin extract of the scrubber seed as an active ingredient, additives commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, Cosmetic compositions can be prepared including conventional adjuvants and / or carriers such as vitamins, pigments and flavorings. In addition, the cosmetic composition may further include a skin absorption promoting substance, in order to enhance the skin whitening effect.

The cosmetic composition for skin whitening of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants -Can be formulated with, but not limited to, a cleansing-containing, oil, powder foundation, emulsion foundation, wax foundation, spray, and the like. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition 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, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. have.

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

If the formulation is a solution or emulsion, a solvent, solubilizer or emulsifier is used as carrier component, for example water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, Fatty acid esters of 1,3-butylene glycol oil, glycerol aliphatic esters, polyethylene glycols or sorbitan.

If the formulation is a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Microcrystalline cellulose; Aluminum metahydroxide; Bentonite; Agar or tracant and the like can be used.

If the formulation is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an 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.

In the cosmetic composition for skin whitening of the present invention, the content of the thin-film skin extract of the scrubber seed may include 0.0001 to 15% by weight, preferably 0.001 to 5% by weight based on the total weight of the composition. If the content of the thin skin coat extract of the loofah S. oi seed is less than 0.0001% by weight, the whitening effect is too weak, and if it exceeds 15% by weight, the increase in the whitening effect due to the increase in content is insufficient and the stability of the formulation is not secured. have.

The thin-film sheath extract of the scrubber seed according to the present invention significantly lowers melanocyte viability than the scrubber meat extract, has a better inhibitory effect on tyrosinase activity in melanocytes, and further reduces melanin content in melanocytes.

Fig. 1 is a diagram illustrating the appearance of the thin-film sheath (A) and the loofah-like flesh (B) surrounding the loofah seed and seed.
Figure 2 is a diagram showing the survival rate of the melanocytes treated with the thin-film envelope extract of the scrubber seed according to the present invention.
Figure 3 is a view of the melanocyte morphology after the treatment of melanin cells with a thin skin coat extract of the loofah S. oi seed in accordance with the present invention under a microscope.
Figure 4 is a diagram showing the effect of the thin-film outer skin extract of the S. gooseum seed according to the present invention on the inhibition of tyrosinase activity in melanocytes.
Figure 5 is a diagram showing the effect of the thin-film shell extract of the Smear cucumber seed according to the present invention on the melanin content in melanocytes.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example  One  : Loofah  Preparation of Thin-film Skin Extracts of Seeds

The white areas surrounding the seedlings of the loofah Soi-oi seeds (the seed bark is black with the shell of the seeds) were peeled off with tweezers. Tweezers were used because the white areas surrounding the seedlings of the scrubber seed were attached to the hands. 1 liter of distilled water was added to 20 g of the thin film skin of the collected Sumi-oi seeds and heated on a low heat of about 50-70 ° C. for 2 hours. The thin film skin of the heated scrubber seed was taken out, and water was removed, and it was powdered into fine powder using a mortar and pestle under liquid nitrogen (about -70 ° C). Put the powder of the thin film skin of the scrubber seed into 1 l of distilled water and treat it for 30 minutes with an ultrasonic cleaner for further dissolution. Then, put the sample obtained by filtration with a filter paper into a freeze-dryer and freeze-dry the thin film of scrubber seed powder. Envelope extract was obtained. The lyophilized fractions were placed in 200 ml of distilled water to treat the cells and treated for 30 minutes in an autoclave to prepare fractions (liquid state) for melanocytes.

The external appearance of the thin-film outer shell (A) and the scumber meat quality (B) which wrap the seed and the seed | soil cucumber are shown in FIG.

Comparative example  One  : Loofah  Preparation of Meat Extract

1 L of distilled water was added to 100 g of scumber meat and heated over a low heat of about 50 to 70 ° C. for 2 hours. The heated scrubber meat was taken out to remove water, and then powdered into fine powder using a mortar and pestle under liquid nitrogen (about -70 ° C). The scrubber meat powder was added to 1 l of distilled water, and then treated with an ultrasonic cleaner for 30 minutes to further dissolve. Then, the sample obtained by filtration with a filter paper was put into a freeze dryer and freeze-dried to obtain a powdery scrubber meat extract.

Experimental Example  One  : Measurement of melanocyte viability

In order to determine the effect of the thin-film sheath extract of the scrubber seed according to the present invention on melanin cell survival, the following experiment was performed.

1. Melanin Cell Culture

Melanocytes (B16F10) used in this experiment were purchased from Seoul National University Cell Line Bank. 500 ml of cell culture medium (dulbecco's modified eagle medium; DMEM) was prepared, and 5 ml of penicillin / streptomycin and 10% FBS (fetal bovine serum) were added thereto. 50 ml of 10% phosphate buffered saline (PBS) was prepared as a medium used for growing (cultured) cells. FBS was dissolved at room temperature to inactivate the remaining complement, and then treated for 30 minutes for thermal inactivation (56 ° C. constant temperature bath). The medium was used after filtration with a 0.2 μm membrane filter. 25 ml of medium was poured into a Petri dish and seeded with melanin B16F10 (1.2 × 10 ⁵ cells). Cell inoculation was performed on a clean bench.

2. Loofah  Treatment of Seed Thin Film Cortex Extract

Well plates were prepared and 500 μl (1 × 10 ⁴ cells contained) of melanocytes (B16F10) were dispensed into each well and incubated in a CO ₂ incubator for 24 hours. 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1, 2, 3, 4, 5 mg of the thin skin coat extract of the scrubber seed prepared in Example 1 and the scrubber meat extract prepared in Comparative Example 1, respectively Each well containing melanocytes was inoculated at a / ml concentration. DMSO was used as a negative control and arbutin (50 μg / ml) was used as a positive control.

3. Melanin cell viability measurement and microscopic observation

1 ml of trypsin-EDTA solution was added to the cell line in culture on the plate, and then reacted at 37 ° C. for 1 minute to separate the cells attached to the plate to recover the cells. 20 μl of MTT solution was added to each well and incubated in a CO ₂ incubator (37 ° C., 5% CO ₂ ) for 4 hours. After incubation, 200 μl of DMSO solution was added and reacted again in a CO ₂ incubator (37 ° C., 5% CO ₂ ) for 1 hour. 1.5 ml of each medium was placed in a centrifuge tube and centrifuged at 13,000 rpm at 4 ° C. The absorbance was then measured at 570 nm with a spectrophotometer. In addition, after 72 hours after the thin skin coat extract of the loofah S. cucumber seed according to the present invention was treated with melanocytes, the morphology of the melanocytes was observed under a microscope.

The survival rate of melanocytes treated with the thin-film sheath extract of the scrubber seed according to the present invention is shown in FIG. 2, and after 72 hours of treatment with the melanin-like thin-film sheath extract of the scrubber seed according to the present invention The result of observing with a microscope is shown in FIG.

As shown in FIG. 2, after 72 hours of treatment with the thin skin coat extract of the Smear cucumber seed according to the present invention on the melanocytes, the melanocyte viability according to the concentration of the thin skin coat extract of the Smear cucumber seed is significantly lowered as the concentration is higher. Confirmed. In addition, it was confirmed that the melanin cell viability of the thin-film envelope extract of the loofah myoi seed according to the present invention was significantly lower than the loofah extract, and it was confirmed that melanin cell death was about 25% or more.

In addition, as shown in FIG. 3, after 72 hours of treatment of the thin-film envelope extract of the loofah S. oi seed according to the present invention on melanocytes, the melanocyte activity of the thin-film envelope extract of the loofah S. oi seed was 2.0 mg / ml. It is observed that this is greatly reduced and the density is greatly reduced. In addition, when the concentration of the thin-film skin extract of the scrubber seed was 5.0 mg / ㎖, it was confirmed that the number of melanocytes in the microscope screen was reduced enough to count.

Therefore, it can be seen that the thin-film sheath extract of the scrubber seed according to the present invention inhibits melanocyte activity.

Experimental Example  2  : In melanocyte Tyrosinase  Active measurement

In order to determine the effect of the thin-film sheath extract of the scrubber seed according to the present invention on the inhibitory effect of tyrosinase activity in melanocytes, the following experiment was carried out.

Specifically, melanin cells (B16F10) (1.2 × 10 ⁵ cells) were dispensed into the culture dish, and the cells were stabilized for 24 hours in a CO ₂ incubator (37 ° C., 5% CO ₂ ). Dispensing the thin-film outer skin extract of the scrubber cucumber seed prepared in Example 1 and the scrubber meat extract prepared in Comparative Example 1 each 0, 0.25, 0.5, 0.75, 1 mg / ㎖ in a culture dish, CO ₂ incubator (37 ° C., 5% CO ₂ ) was incubated for 72 hours. 1 ml of trypsin-EDTA was added thereto, followed by reaction at 37 ° C. for 1 minute to separate the cells attached to the medium to recover the cells. 150 μl of 1 M phosphate buffer (pH 6.8) and 20 μl of 3 mM L-tyrosine solution were added. The reaction was started by adding 10 μl of 2,500 U / ml mushroom-derived tyrosinase. At this time, 0.2 μM α-MSH reagent (melanocyte stimulating hormone, melanocyte stimulating hormone, 0.5ml) was added to investigate whether the reaction was positive. The cells were incubated at 37 ° C. for 30 minutes, and the absorbance was measured at 405 nm after 1 hour. Distilled water was used as a negative control group, and arbutin (50 µg / ml) was used as a positive control group.

The results are shown in Table 1 and FIG. 4.

Tyrosinase Activity in Melanocytes with Different Concentrations of Thin Film Skin Extracts from α-MSH Tyrosinase activity Comparative Example 1
(Scenic cucumber meat extract)
(Mg / ml) 0 0.2 μM 2.4 0.25 0.2 μM 2.34 0.5 0.2 μM 2.11 0.75 0.2 μM 1.86 One 0.2 μM 1.55 Example 1
(Thin-skinned Skin Extract of Smear-Oil Seeds)
(Mg / ml) 0 0.2 μM 2.42 0.25 0.2 μM 2.07 0.5 0.2 μM 1.86 0.75 0.2 μM 1.8 One 0.2 μM 1.5 Voice Control Group (DMSO) 0 One Positive control group (arbutin) (50 µg / ml) 0.2 μM 1.35

As shown in Table 1 and Figure 4, it was confirmed that the thin-film sheath extract of the loofah S. cucumber seed according to the present invention has a better inhibitory effect on tyrosinase activity than the loofah extract.

Experimental Example  3  : Determination of melanin content in melanocytes

In order to determine the effect of the thin-film sheath extract of the scrubber seed according to the present invention on the melanin content in the melanocytes, the same procedure as in Experiment 2 was carried out to measure the melanin content in the melanocytes.

The results are shown in Table 2 and FIG.

Melanin Contents in Melanocytes According to Concentrations of Thin Film Skin Extracts α-MSH Melanin content Comparative Example 1
(Scenic cucumber meat extract)
(Mg / ml) 0 0.2 μM 2.45 0.25 0.2 μM 2.25 0.5 0.2 μM 2 0.75 0.2 μM 1.94 One 0.2 μM 1.62 Example 1
(Thin-skinned Skin Extract of Smear-Oil Seeds)
(Mg / ml) 0 0.2 μM 2.47 0.25 0.2 μM 2 0.5 0.2 μM 1.81 0.75 0.2 μM 1.79 One 0.2 μM 1.55 Voice Control Group (DMSO) 0 One Positive control group (arbutin) (50 µg / ml) 0.2 μM 1.35

As shown in Table 2 and Figure 5, it was confirmed that the melanin content in the melanocytes treated with the thin skin coat extract of the loofah myoi seed according to the present invention reduced the melanin content more than the group treated with loofah myi meat extract.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation example  One  : Flexible cosmetics (content: weight%)

Thin-film Skin Extract of Smear Cucumber Seeds 0.01

Glycerin 3.0

Butylene Glycol 2.0

Propylene glycol 2.0

Carboxy Vinyl Polymer 0.1

Ethanol 10.0

Triethanolamine 0.1

Preservatives, trace pigments, trace fragrances, trace amount of purified water

Total 100.0

Formulation example  2  : Nourishing lotion (content: weight%)

Thin-film Skin Extract of Smear Cucumber Seeds 0.01

Beeswax 4.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.5

Liquid Paraffin 5.0

Squalane 5.0

Caprylic / Capric Triglycerides 5.0

Glycerin 3.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Carboxy Vinyl Polymer 0.1

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances, trace amount of purified water

Total 100.0

Formulation example  3  : Nutritional Cream (Content: Weight%)

Thin-film Cortex Extract of Loofah Seeds 0.005

Beeswax 10.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.5

Liquid Paraffin 10.0

Squalane 5.0

Caprylic / Capric Triglycerides 5.0

Glycerin 5.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances, trace amount of purified water

Total 100.0

Formulation example  4  : Massage  Cream (Content: Weight%)

Thin-film Cortex Extract of Loofah Seeds 0.005

Beeswax 10.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.8

Liquid Paraffin 40.0

Squalane 5.0

Caprylic / Capric Triglycerides 4.0

Glycerin 5.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances, trace amount of purified water

Total 100.0

Formulation example  5  : Pack (content: wt%)

Thin-film Cortex Extract of Loofah Seeds 0.005

Polyvinyl Alcohol 13.0

Sodium Carboxymethylcellulose 0.2

Allantoin 0.1

Ethanol 5.0

Nonylphenyl Ether 0.3

Preservatives, trace pigments, trace fragrances, trace amount of purified water

Total 100.0

Claims (3)

A cosmetic composition for skin whitening containing a thin film skin extract of Smear cucumber seed as an active ingredient. The cosmetic composition for skin whitening according to claim 1, wherein the content of the thin-film sheath extract of the loofah seed is contained in an amount of 0.0001 to 15 wt% based on the total weight of the composition. The skin of claim 1, wherein the composition is prepared in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder. Whitening cosmetic composition.

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