KR100899847B1 - Cosmetic compositions for skin-whitening - Google Patents

Abstract

The present invention relates to a whitening cosmetic composition containing Yangchun white snow has a skin lightening effect.

Composition, whitening cosmetic, Yangchun white snow

Description

Cosmetic Compositions for Skin-Whitening Containing Extracts of Baekbong-Ryeong, Hemp, Yeonja, and Gumsil {Cosmetic Compositions for Skin-Whitening}

The present invention relates to an extract having a whitening effect, and relates to a cosmetic composition exhibiting an excellent whitening effect when Baekbongyeong, hemp, softwood, and gumsil mixed extracts are mixed into a formulation.

   Human skin color is determined by the amount of melanin, carotene and hemoglobin, of which melanin is the most decisive factor. Melanin is synthesized in melanocytes (melanocytes), which are pigment cells in the basal layer of the skin (epidermis), and are transferred to peripheral keratinocytes (Keratinocytes) to represent human skin color. An abnormally low production of melanin causes skin lesions such as vitiligo. Overproduction, on the contrary, forms one of the main concerns among middle-aged women, blemishes and freckles, and is also closely related to skin cancer (Melanoma).

   The whitening action is described in more detail as follows. Melanin is produced by tyrosine (Tyrosine) through the enzyme and non-enzymatic oxidation reaction in the cell called melanocytes in the basal layer of the skin epidermis, and is transferred to the keratinocytes constituting the epidermis. Important enzymes involved in the biosynthesis of melanin include tyrosinase, tyrosinase-related protein 1 and dopachrome tautomerase. Two enzymes besides tyrosinase have been recently developed and recently attracted attention, but the enzyme that plays a decisive role in the synthesis of melanin is tyrosinase. The first stage of biosynthesis of melanin is caused by tyrosinase. Tyrosinase causes tyrosine to be dopa (Dopa), which makes it dopaquinone (Dopa-quinone). The next steps of these two reactions are known to be possible by non-enzymatic reactions, and tyrosinase alone is known to produce melanin.

   Recently, a new theory of melanin formation following the expression of signal transduction agents in skin cells has been proposed. That is, when the skin is exposed to ultraviolet light, interlukin-alpha, which causes inflammation, is expressed from keratinocytes in the epidermis, and the interleukin-alpha expresses keratinocyte endothelin. To promote. This series of processes increases the formation of melanocytes and activates melanocytes. In the melanoma in the melanocytes thus formed, melanin is formed by tyrosine tyrosine.

   Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A 6-192062), kojic acid (JP-A 56-7710), arbutin (JP-A 4-9315) and some compounds already have tyrosinase inhibitory activity and whitening. Although it is used as a raw material for cosmetics, its use is limited due to poor stability in the prescription system, discoloration, discoloration, off-flavor, efficacy at the biological level, unclear effect and safety issues. And the inhibitory effect of tyrosinase has been demonstrated, but the effect is low in experiments similar to the actual biologic level. Therefore, the inhibitory effect of the melanoma cell culture similar to the biological level is required. Hydroquinone is also regulated as a carcinogenic substance and is prohibited from use. Kojic acid and ascorbic acid are very unstable substances. Browning can occur when stored in small amounts for several weeks at room temperature. Plant natural products are not only safe but also have many beneficial ingredients, making them a good candidate for whitening.

   In order to solve the problems of existing raw materials with whitening effect by suppressing melanin production for a long time, various kinds of abnormal melanin production can be suppressed from high-efficiency plant extracts such as herbal medicines, vegetables, fruits, and flowers. Efforts have been made to find extracts that have an effect.

Poria cocos. (Schw.) Wolf.It grows like a hump from the roots of pine trees many years ago, and the white one is called Baekbok-ryeong, and the pink one is called Red Bok-Ryeong.

As a medicinal crop with medicinal effects, it was used as a medicinal herb rather than as a food. According to Dongbogam, the taste of Baekbokyeong is sweet, boring, and flat. Thus, it has been used a lot as a tonic since ancient times and also used for recovery after illness. The main component of Bokryeong contains β-Pachyman and accounts for about 93% of the dry weight. Triterpenoid compounds contain Pachymic acid, Tumulosic acid, 3-β-Hydroxylanosta-7, 9 (11), and 24-trien-21-oic acid. In addition, it contains Chitin, Protein, Fat, Sterol, Lecithin, Glucose, Adenin, Histidine, Choline, β-Pachyman Degrading Enzyme, Lipase and Protease. It is excellent for cancer suppression, stomach, tonic, pancreas and kidney.

It has been used as a tonic since ancient times. It works on menopause, parenteral, heart, nerve, and bladder to protect the spleen, sputum, and stabilize the mind. The results of pharmacological experiments have shown that diuretic, lowering blood sugar and sedative effects are also known as immune resuscitation.

Hemp ( Dioscorea opposita Thunb) is a perennial herb. Roots are large, clumped columnar with triangular oval leaves facing each other. Stems and leaf stalks are branched.

It is said that Chinese soldiers of the Spring and Autumn Chuo period used large roots of white-flowering plants as food in the summer. It was called `` sanwoo '' because it happened by chance to find food in the mountains. Later, it was used as a good medicine for fire extinguishers, and it was renamed as `` acid medicine '' (山藥).

The main components of hemp are Dioscin, Batatasin, mucin mucin, Amylase and other digestive enzymes, amino acids, and met.

The merits of the town can be summarized in two ways: health and health (健胃), Gangjeong action. Stomach action is to enhance the digestive system functions such as stomach and spleen to enhance the appetite and digestion. Strong action to heal the collapse of the whole body. Symptoms associated with stagnation weakness, urinating, urinary strength, endless symptoms, cysts, impotents and remarkably improves.

Nelumbo nucifera is elliptical or round in shape, 1.2-1.8 cm long and 0.8-1.4 cm in diameter. The surface is pale yellowish brown or reddish brown with papillary papillae at the center of one end and dark brown. The vagina is hard, not easily split, and the skin is thin but does not fall well. The taste is sweet and salty and the property is flat without being biased in either direction. Seeds of the fruit include methylcorridin dimethyl cocurin, N-nornousiferin, nroomumbin, nufarin, armephabin, rotucin, liensinin, isoriensinin and neferin. It is a medicine that has been widely used because it helps the stomach and the spirit and energy, and when it is taken for a long time, it makes the body light, not old, hungry, and has a long life. It acts on the spleen, kidneys and heart. It strengthens the function of the spleen to stop diarrhea and boosts the function of the kidneys, and acts on the heart to stabilize the mind.

Euryale ferox Salisb is dried seed of spiny lotus belonging to the water lily family. It tastes sweet and does not occupy the nature. It acts to protect the spleen and kidney. The thorn lotus is planted in the ponds of the sub-central part of Korea, and it picks up ripe fruits in autumn and cuts the hard bark and dry it in the sun. Seeds contain a lot of starch, and the ingredients in 100g are 4.4g protein, 0.2g fat, 32g carbohydrate, 0.4g fiber, 0.5g ash, 9mg calcium, 110mg phosphorus, 0.4mg iron, vitamin B, vitamin C, It contains trace amounts of nicotinic acid and carotene. Strengthen kidney function, and strengthens rain, gastrointestinal, and digestive function. Relieves symptoms caused by stress. Encourage 精 氣 (regular) to improve memory and brighten eyes and ears. Taking it for a long time can make your body lighter, less hungry, not old and fresh.

   Accordingly, it is an object of the present invention to provide a plant natural extract having excellent skin whitening effect by inhibiting the activity of tyrosinase enzyme and inhibiting melanin production and having no safety problems.

   Another object of the present invention is to provide a cosmetic composition comprising an extract having such efficacy.

   In view of the present invention, the present inventors search for an effective substance for whitening among various natural products of nature in order to find an essential substance having a skin whitening effect, Baekbong-ryeong, hemp, soft and mixed extracts (Yangchun white snow) It was found to show excellent whitening effect and solved stability problem at the same time.

 In addition, the cosmetic composition was found to exhibit a clear whitening effect even in the experimental results of applying directly to the skin of the Yangchun white snow snow at a certain concentration. The present invention is to provide a whitening cosmetic composition containing Yangchun white snow that can fundamentally prevent abnormal melanin formation.

 This extract is added to cosmetics, that is, softening water (skin), nourishing lotion (lotion), nourishing cream, massage cream, essence, packs and the like.

The present invention relates to a whitening cosmetic composition containing Yangchun white snow has a skin lightening effect. That is, it relates to a whitening cosmetic composition, characterized in that it comprises a Baekbongryeong, hemp, softwood, gumsil extract (hereinafter Yangchun Baekseolgo).

As can be seen in the experimental example, Yangchun white snow of the present invention has an excellent inhibitory effect on tyrosinase activity and melanin production, cosmetics prepared by the addition of the extract of the present invention was excellent skin whitening effect.

 The present invention is described in more detail as follows. One or more solvents selected from the group consisting of distilled water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, hydrous butylene glycol, hydrous propylene glycol and hydrolysed glycerin are extracted as dried solvents. Add 5-10 times by volume. Extraction method is a method of extracting by heating at 50-100 ℃ for 2-20 hours or extracting the active ingredient at 5-37 ℃ for 1-15 days while the cooling condenser is installed to prevent the solvent from evaporating. Can be. The extracted Yangchun Baekseol is concentrated under reduced pressure while recovering the solvent evaporated using a distillation apparatus equipped with a cooling condenser, and then added to the cosmetic in an amount of 0.001-10%, preferably 0.01-5%, as its dry weight. .

   The following Examples and Experimental Examples illustrate the content of the present invention, but the content of the present invention is not limited thereto.

<Example 1>

    500g of dried Baekbong-ryeong, hemp, softwood, and gumsil were placed in 5kg of distilled water and extracted by heating at 70 ℃ for 3 hours using an extractor equipped with a cooling condenser. After standing and aged, the product was filtered with a 0.45 μm filter. The filtrate was completely removed from the solvent in a vacuum rotary evaporator to obtain 15.1 g of an extract.

menstruum Yield (kg) Example 2. Water Butylene Glycol (30 w / w%) 4.1 Example 3. Hydrous Propylene Glycol (30 w / w%) 4.1 Example 4. Hydrous ethanol (30 w / w%) 4.4 Example 5. Hydrous ethanol (70 w / w%) 4.3 Example 6. ethanol 4.2 Example 7. Methanol 4.3 Example 8. Butylene glycol 4.0 Example 9. Propylene glycol 4.1 Example 10. glycerin 3.9

Experimental Example 1. Measurement of cytotoxicity

Cytotoxicity in monolayer cell cultures was measured for Yangchunbaek Snow.

   Experiment method

MTT assay is a method widely used for cell proliferation or toxicity by measuring the number of living cells. The living cells are water-soluble and yellow salts of MTT (3- [4,5-dimethylthiazole-2-yl] -2,5 in the mitochondria. It is based on the principle that -diphenyltetrazolium bromide is reduced to a water-insoluble blue formazan derivative by succinate dehydrogenase (also called succinate dehydrogenase or mitochondrial dehydrogenase). The resulting formazan derivative is dissolved by adding a solubilizer (usually dimethylsulfoxide) and then measured for absorbance. In more detail, human fibroblasts cell line (ATCC, CRL-2310) was inoculated in 24well plates at a concentration of 1x10 ⁴ cells / ml. Medium was used DMEM (Dubelcco'S Modified Eagle Medium, BRL, USA) containing 10% bovine serum. 24 hours after the inoculation, DMEM (Dubelcco'S Modified Eagle Medium, BRL, USA) containing 2% of bovine serum was replaced, and 10 μl of the test sample was added at a concentration of 50 μg / ml, followed by 37 ° C. and 5% for 3 days. Incubated in a CO ₂ incubator. After incubation, remove the supernatant and wash again with 200ul of 5% PBS (phosphate buffered saline), add 1.0ml of MTT solution per well, remove MTT after 4 hours, and add 1.0ml of DMSO per well overnight. Absorbance was measured at 570 nm after incubation at ℃. The results are shown in Table 1.

Cytotoxicity in Monolayer Cell Culture of Yangchun Baekseol  Example Cell survival rate (%) Example 1 101.2 Example 2 102.5 Example 3 100.9 Example 4 103.2 Example 5 104.7 Example 6 100.7 Example 7 101.9 Example 8 101.4 Example 9 100.2 Example 10 103.1

Experimental Example 2. Free radical scavenging activity of Yangchunbaek snow

For the Yangchun-baek snowy snow, using DPPH (1,1-Diphenyl-2-picrylhydrazyl radical) according to Park's method (Park, S. N, Journal of Korean Society of Cosmetic Products, 23 (1) , pp75-132, 1997) The radical scavenging effect was measured.

To 1 ml of 0.2 mM DPPH (1,1-diphenyl-2-picrylhydrazyl) solution, 2 ml of Yangchun white snow sugar was added at each concentration, reacted at room temperature for 10 minutes, and the absorbance was measured at 520 nm. (%) Was obtained. At this time, as a control group, methanol was used instead of DPPH, and 2 ml of Yangchunbaeksulgo was added at the same concentration as the experimental group. In addition, as a positive control of the free radical scavenging ability, it was compared with the green tea extract containing a large amount of flavonoid components excellent in the free radical scavenging ability.

Free radical scavenging activity SC ₅₀ was expressed as the concentration (μg / ml) of Yangchun Baekseolgo required to remove 50% of the free radicals.

Looking at the results of the following Table 2, the Yangchun Baekseolgo of the Example compared to the free radical scavenging ability of the green tea extract, it was confirmed that the free radical scavenging ability by showing the SC ₅₀ concentration at a similar or lower concentration.

Free radical scavenging effect of Yangchunbaek snow Example SC ₅₀ (μg / ml) Example 1 56 Example 2 69 Example 3 32 Example 4 47 Example 5 57 Example 6 48 Example 7 52 Example 8 49 Example 9 54 Example 10 37 Green Tea Extract 45

Experimental Example 3. Inhibitory Effect of Yangchun Baekseol on Intracellular Tyrosinase Activity

The inhibitory activity of tyrosinase was measured on the Yangchun-baeksago. Well counts according to all concentrations were prepared in triplicate. B16 melanoma cells (ATCC CRL6323) were seeded in Dulbecco's modified Eagle's Medium (DMEM) with 10% bovine serum and 1% antibiotic at a density of 1 × 10 ⁵ cells and incubated at 5% CO ₂ , 37 ° C. for one day. Thereafter, the cells were treated with DMEM 10% medium treated with α-MSH (melanocyte stimulating hormone; melanocyte stimulating hormone, 0.34 ug / mL, Sigma), and the samples of the examples were incubated for 3 days. Remove the medium and remove the cells by trypsin treatment, put in a 1.5mL tube and centrifuged for 5 minutes at 12,000rpm to discard the supernatant and wash twice with PBS. To the cell precipitate, 1 ml of 80% potassium phosphate buffer solution (pH6.8) containing 1% Triton X-100 and 2 mM PMSF (Phenylmethylsulfonyl Fluoride) was added, and the mixture was shaken for 20 minutes on ice for 1 hour at 4 ° C. Leave it for a while. Then, the supernatant is collected by centrifugation at 4 ° C., 20 minutes, and 12,000 rpm. 0.3 mL of the supernatant and 0.3 mL of 1.5 mM tyrosine are reacted at 37 ° C for 3 hours. The mixture was vigorously shaken and the absorbance was measured at 475 nm. Protein quantification of the supernatant was analyzed according to the Bio-rad protein quantification protocol, the absorbance of the resulting melanin was divided by the amount of protein. The extract of Morus bark used as a control has a good tyrosinase activity inhibitory effect and is widely used as a cosmetic raw material.

 % Tyrosinase activity inhibition = [(A-B) / A] x100

       A: Absorbance at 475 nm of reaction solution without sample

B: absorbance at 475 nm of the reaction solution to which the sample was added

Inhibitory Effect of Yangchun Baekseol on Tyrosinase Activity sample Tyrosinase inhibition rate (%) 50 μg / ml 100 μg / ml Example 1. 35.2 43.8 Example 2. 53.6 72.8 Example 3. 42.7 54.9 Example 4. 37.3 43.9 Example 5. 39 49.1 Example 6. 42 49.5 Example 7. 47.8 55.9 Example 8. 36.2 45.1 Example 9. 48.2 58.6 Example 10. 46.8 63.1 Lettuce extract 33.1 42.5

Experimental Example 4. Experiment of melanin production inhibition using B16 melanoma of Yangchunbaek Snow

   The effects of B16 melanoma on melanin production were measured in Yangchunbaek Snow.

Melanogenesis inhibitory function was assayed at the cellular level through B16 melanoma cell culture. B16 melanoma cells were incubated in Dulbecco's modified Eagle's Medium (DMEM) containing 10% bovine serum at 5% CO ₂ , 37 ° C for 2 days until 1.0 X 10 ⁸ cells / T25 flask. Then, put Yangchun Baekseol and control samples by concentration and incubated for 2 days at 5% CO ₂ , 37 ℃. Then trypsin is added to collect the cells and observe the degree of blackening of the cells. Melanin was quantified by measuring the absorbance at 475 nm, taking the supernatant of melanin-produced and discharged T25 flask, and counting the cells in a ratio of collected cells: tryphan blue = 1: 9 Count the amount of melanin produced per unit cell to show the inhibitory effect. The extract of Morus bark used as a control has a good tyrosinase activity inhibitory effect and is widely used as a cosmetic raw material.

Melanin production = extinction coefficient (0.1498) × A ₄₇₅

Melanin production inhibitory effect = 1-melanin production / sample cell count

Inhibitory Effects of Melanin Biosynthesis on B16 melanoma Example Melanin biosynthesis inhibitory effect (%) 50 µg / ml 100 µg / ml Example 1 23 32 Example 2 48 69 Example 3 35 47 Example 4 29 38 Example 5 25 32 Example 6 17 39 Example 7 22 28.7 Example 8 28 21.3 Example 9 19.7 25 Example 10 27.5 48.6 Morus Extract 38 53

< Prescription Example 1>

   A prescription example of supple cosmetics (skin) among cosmetics containing Yangchun white snow is as follows. Yangchun white snow here is that of Example 2. Comparative Example 1 is a formulation in which an equal amount of 1,3-butylene glycol is added instead of Yangchun Baekseolgo.

Ingredient Prescription Example 1 (% by weight) Comparative Example 1 (% by weight)  Yangchun white snow 2.0 - 1,3-butylene glycol 2.00 4.00 glycerin 3.00 3.00 Phenyltrimethicone 2.00 2.00 Fiji-60 Hydrogenated Castor Oil 0.5 0.50 ethanol 6.00 6.00 antiseptic a very small amount a very small amount incense a very small amount a very small amount Purified water Remaining amount Remaining amount system 100.00 100.00

< Prescription Example 2>

   A prescription example of milk lotion among the cosmetics containing Yangchunbaek snow is as follows. Yangchun white snow here is that of Example 2. Comparative Example 2 is a formulation in which the same amount of 1,3-butylene glycol was added instead of Yangchun Baekseolgo.

Raw material Prescription Example 2 Comparative Example 2   Yangchun white snow 5.0 - 1,3-butylene glycol 4.00 9.00 glycerin 3.00 3.00 Cetearyl Alcohol 0.50 0.50 Polysorbate 20 1.00 1.00 antiseptic a very small amount a very small amount incense a very small amount a very small amount Purified water Remaining amount Remaining amount system 100.00 100.00

< Prescription Example 3>

A prescription example of nourishing cream among cosmetics containing Yangchun Baekseol is as follows. Yangchun white snow here is that of Example 2. Comparative Example 3 is a formulation in which the same amount of glycerin was added instead of Yangchun Baekseolgo.

Raw material Prescription Example 3 Comparative Example 3   Yangchun white snow 5.0 - glycerin 5.00 8.00 Triethanolamine 0.50 0.50 Tocopheryl acetate 1.00 1.00 Hydrogenated Polydecene 5.00 5.00 Squalane 5.00 5.00 Macadamia Nut Oil 12.00 15.00 Polysorbate 60 1.00 1.00 Sorbitan sesquioleate 1.00 1.00 Carboxy vinyl polymer 0.20 0.20 antiseptic a very small amount a very small amount incense a very small amount a very small amount Purified water Remaining amount Remaining amount system 100.00 100.00

< Prescription Example 4>

A prescription example of a massage cream among cosmetics containing Yangchunbaek snow is as follows. Yangchun white snow here is that of Example 2. Comparative Example 4 is a formulation in which the same amount of glycerin was added instead of Yangchun Baekseolgo.

Raw material Prescription Example 4 Comparative Example 4 Yangchun white snow 2.0 - Vaseline 5.00 5.00 Hydrogenated Polydecene 30.00 30.00 Beeswax 3.00 3.00 Polysorbate 60 1.00 1.00 Sorbitan sesquioleate 1.00 1.00 1,3-butylene glycol 5.00 5.00 glycerin 5.00 7.00 antiseptic a very small amount a very small amount incense a very small amount a very small amount Purified water Remaining amount Remaining amount system 100.00 100.00

Test Example 1. Evaluation of the whitening effect

< Evaluation of Whitening Effect at Human Skin Level>

   Twenty healthy men and women affixed an opaque tape with six holes of 1.5 cm in diameter on the upper forearm of the subject, and then treated them with 1.5 ~ 2 times of ultraviolet light (MED; Minimal Erythema Dose). UVB) was irradiated to induce pigmentation of the skin, and two months after application of the test substances, the contrast of the skin was measured using a colorimeter. Each test substance was applied to prescription Examples 1 to 3 and Comparative Examples 1 to 3 twice daily over morning and evening.

The determination of the effect was determined by obtaining an L ^* value representing the contrast of the skin. In general, the L ^* value of Korean skin that does not induce artificial pigmentation is in the range of 50 to 70.

If the test substances are applied, the L ^* value will gradually increase. The difference in skin color (ΔL ^* ) between the start point and the end point of application was calculated according to the following equation, and the whitening effect on the human skin was compared. The measurement results are shown in Table 5.

ΔL ^* = ΔL ^* value on the last day of test substance application-L ^* value before test substance application

Skin contrast measurement result Test substance Whitening Effect (ΔL ^* ) Prescription Example 1 3.93 Comparative Example 1 1.20 Prescription Example 2 5.12 Comparative Example 2 1.56 Prescription Example 3 4.83 Comparative Example 3 0.94

As a result of the investigation, it can be seen that the skin whitening effect is superior to Comparative Examples 1 to 3 prepared without adding the cosmetics of Formulas 1 to 3 prepared by adding Yangchun white snow.
The whitening cosmetic composition is preferably one having a formulation selected from a lotion (skin lotion), nutrition lotion, nutrition cream, massage cream, essence, pack.

Claims (5)

A whitening cosmetic composition containing an extract of Baekbong-ryeong, hemp, softwood, and gumsil having a skin whitening effect. The method of claim 1, Whitening cosmetic composition, characterized in that the extraction solvent is distilled water, methanol, ethanol, propanol, butanol, ethyl acetate, acetone, hexane, propylene glycol, butylene glycol, glycerin alone or a mixture of two or more. The method of claim 1, Baekbong-ryeong, hemp, yeonja, and gumsil extracts were added to the extraction of 5-10 times the volume of the extraction solvent with respect to the weight ratio of dried Baekbong-ryeong, hemp, yeonja, and gumsil, and then 50-100 ° C. using an extractor equipped with a cooling condenser. Whitening cosmetic composition, characterized in that the extraction by heating for 2-20 hours or by extracting the active ingredient by deposition at 5-37 ℃ for 1-15 days. The method of claim 1, Whitening cosmetic composition, characterized in that the extract of the Baeknyeongyeong, hemp, soft and gumsil is contained in an amount of 0.001-10% as dry weight, 0.01-30% as an extract to the total weight of the whitening cosmetic composition. The method of claim 1, The whitening cosmetic composition is a whitening cosmetic composition, characterized in that it has a formulation selected from a lotion (skin lotion), nutrition lotion, nutrition cream, massage cream, essence, pack.