KR20020035656A - Inhibition agent of tyrosinase composed of saururus chinensis(lour) baill extract and cosmetic composition having whitening effect containing the same - Google Patents

Abstract

PURPOSE: Provided are an inhibition agent of tyrosinase composed of Saururus chinensis(lour) baill extract and a cosmetic composition having whitening effects and containing the same. The extract of Saururus chinensis(lour) baill is excellent in tyrosinase inhibition and whitening effects. CONSTITUTION: The extract of Saururus chinensis(lour) baill is prepared by the steps of: adding water or a mixed solvent of water and at least one selected from methanol, acetone, propanol, ethanol, 1,3-butylglycol and propyleneglycol, in the amount of 6-12 L to 1 kg of Saururus chinensis(lour) baill; extracting it at 60-80 deg.C for 6-12 hours; and filtering the extract then maturing it at 1-30 deg.C for 5-10 days. The cosmetic composition contains 0.0001-20 wt.% of the extract to dried weight of cosmetic composition.

Description

Tyrosinase inhibitor consisting of triticale extract and whitening cosmetics containing the same {INHIBITION AGENT OF TYROSINASE COMPOSED OF SAURURUS CHINENSIS (LOUR) BAILL EXTRACT AND COSMETIC COMPOSITION HAVING WHITENING EFFECT CONTAINING THE SAME}

The present invention relates to a whitening cosmetics containing tritical herb extract extracted from tritical plant.

Human skin color is generally determined by the thickness of the skin, 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 present in the basal layer of the skin (epidermis), and are transferred to peripheral keratinocytes (Keratinocytes) to represent human skin color.

Melanocytes are cells of neural capacity that produce melanin and play an important role in protecting the skin from UV rays. Melanocytes are morphologically similar to neurons, such as dendritic protrusions, and have many receptors for signaling agents, growth factors, etc., and thus have many characteristics that show their developmental origin as neurons.

Melanocytes are affected by various factors such as hormones and external conditions such as ultraviolet rays and inflammation. In addition, in order to produce melanin from melanocytes, an enzyme called tyrosinase acts as a substrate using tyrosin in a cell to generate dopaquinone, and spontaneous and enzymatic reactions from dopaquinone Melanin, a black pigment, is produced.

When abnormally low melanin is produced, skin lesions, such as vitiligo, are induced. In contrast, overproduction produces melanoma, blemishes, freckles, and skin cancer (Melanoma).

Therefore, ascorbic acid, kojic acid, arbutin, hydroquinone have been conventionally used to reduce the symptoms of abnormal skin pigmentation such as blemishes, freckles, pigmentation, and excessive melanin pigmentation caused by exposure to ultraviolet light. Although substances with tyrosinase inhibitory activity, such as, have been formulated and used in cosmetics and medicines, ascorbic acid has a relatively low inhibitory effect on tyrosinase activity and low stability of the molecule itself, and thus it is not suitable as a whitening raw material, and kojic acid is applied to tyrosinase active sites. While chelating to the contained copper ions to inhibit the activity of the enzyme is excellent in performance, but because the oxidation reaction is caused by light or oxygen, there is a lot of stability problems are not suitable as a cosmetic.

In addition, hydroquinone is used locally for the treatment of hyperpigmentation such as blemishes, freckles, spots and hyperpigmentation, but it has a strong melanogenesis inhibitory activity, while causing denaturation or lethality of pigment cells and their intrinsic function. Since side effects, such as damage, appear, it is forbidden to use in cosmetics.

As described above, conventional substances having tyrosinase inhibitory activity have been used in cosmetics and medicines, but as mentioned above, such factors as safety problems on the skin, formulations in cosmetics or medicines, insufficient stability and whitening effects, etc. Its use is limited.

In addition to these conventional whitening agents, research has been conducted to find whitening active ingredients in natural products, especially plants. Among them, lettuce skin (Japanese Patent Publication No. 55-44375, Small 64-26507, Small 64-83009, Flat 1) 256587), Licorice (Japanese Patent Laid-Open No. 60-214721, Small 63-23809, Small 64-63506, Hei 1-49706), Peony (Japanese Laid-Open Patent No. 61-246109), Cinnamon (Japanese Laid-Open Patent No. 63-30403) It has been found that a number of plant extracts act on tyrosinase to inhibit melanin production, but they also have many problems to be used above the effective concentration in cosmetics or pharmaceuticals in terms of safety and discoloration potential.

On the other hand, three hundred seconds is a perennial herb belonging to the three hundred genus (Saururaceae), three hundred seconds (Saururus), the upper part of this plant stem is called three hundred seconds (三 白草) because two to three leaves are white.

Three hundred (Saururaceae) is distributed in Korea, Japan, China, North America, Mexico, etc. and classified into five genus 7 species as follows [Wu.C.Y., Acta Phytotaxon, Sinica, 6, 222 (1957)].

Fam. Saururaceae Genus Anemopsis… A. Californica Genus Saururus … S. Cernuus S.Chinensis Genus Houttuynia… H.Cordata Genus Gymnotheca… G. chinensis G. involucrata Genus Circaeocarpus… C. Saururoides

Among these, three native plants ( Sarurus chinensis Baill) and wild wheat ( Houttuynia cordata Thunb) grow wild in Korea. These are all perennial plants that grow in wetlands, with three hundred vines growing on Hyeopjae, Jeju Island, and Ulleungdo, Jeju Island, and the central region of Jeju Island. Is grown.

In the Chinese Medicine Dictionary (Shanghai Science and Technology Publishing House), the three hundred seconds are the tree trunk, Orobaek, Samgebaek, Baekhwayeon, Jeonsambaek , Cheonseongcho (天性 草) is another name. In addition, the three hundred seconds are 30 ~ 90 cm in height, the stem is upright, or the lower side obliquely, the underground stem extends to the side. The leaves are regenerated, egg-shaped or ovate lanceolate, pointed at the end, and lobes are present.

Three hundred seconds in oriental medicine has been used for detoxification, wind poison, stool, hypertension, diuresis, and is also used for the treatment of customary arthritis, such as underwear or external use. Its main ingredients include Quercetin, Quercitrin, Isoquercitrin, and Hyperin in the leaves, and methyl-n-nonylketone (Essential oil) in the outpost. Methyl-n-nonyl ketone) contains rutin in the stem (Chinese medicine dictionary). These quercetin, quelcitrin, isocuelcitrin, hyphenin, rutin and the like are flavonoids (flovonoides), flavonoids such as polyphenols (polyphenols) have an antioxidant action, that is, free radical scavenging action. Zheng et al. Presented flavonoid assays in the outpost. (徐 (, 沙 世 炎: 中草藥 有效 成分 分析 法: 下 冊, 東京, 34 (1984))

Recently, three hundred seconds of pharmacological studies have identified dimethyl terephthalate and quercetin isolated (Kwak Jae-wook, Kyung Hee University, DP518.14-Kwak853) and three hundred seconds above ground components. A study (Lee Sung-woo, Seoul National University, DM 518.19- Lee 381s) is known, and Korean Patent Application Publication No. 96-21001 describes an example of using it as a cosmetic for preventing aging by using its radical scavenging action. Recently, degenerative diseases related to aging and adult diseases such as atherosclerosis, high blood pressure, diabetes, etc. are becoming a social problem, and the oxygen harmful theory is gradually recognized that the cause is caused by free radicals. ((Halliwell, B .: Drug oxidant effects.Drugs 42,569 (1991); Kawasaki, Japan): As an antioxidant. Japan 46,2286 (1988)]

Free radicals are chemically highly reactive chemicals that occur when the skin is irradiated with UV light or during the respiration of cells in the skin. Most of the free radicals associated with aging of the skin are reactive oxygen species, which include superoxide radicals, hydrogen peroxide, hydroxy radicals, and singlet oxygen. The reactive oxygen species are generated during ultraviolet irradiation or in the respiratory process to cause peroxidation of lipids, which are components of cell membranes, through a multi-step chain reaction. Lipid peroxidation also generates various active species such as radicals, aldehydes, epoxides, and damages DNA, RNA, proteins, cell membranes, and cellular structures. Toxicity of these reactive oxygen species is considered to be a major cause of cancer, tissue damage, and aging. (Black HS, Photochem potobiol, 46 (2), 213, 1987) Although aging and melanin production have a common factor due to UV irradiation, their origins are different and they are considered to be UV only. Not only that, but the correlation between aging and melanin is still unknown.

The present inventors have studied steadily in search of a superior tyrosinase activity inhibiting agent in view of the above problems associated with the use of conventional whitening efficacy substances or plant extracts having tyrosinase inhibitory activity in cosmetics and pharmaceuticals. In particular, in order to solve the safety problem, the active substance was searched for natural plants that have been used for herbal or folk remedies for a long time has proven its safety. As a result, it was found that the extract of the genus Triticales exhibits a very excellent tyrosinase activity inhibitory effect in addition to the known effects, to complete the present invention.

1 is a graph showing the inhibitory effect of tyrosinase activity of the three hundred sec extract and Arbutin obtained in Example 2.

Therefore, according to the present invention, there is provided a tyrosinase inhibitor consisting of three hundred sec extract, and further, a whitening cosmetic containing the extract is provided.

Hereinafter, the content of the present invention in more detail as follows.

In the present invention, three hundred seconds, S. genus 300 seconds. S. Chinensis spp. And S. S. Cernuus species can be used in all, but since S. cernus grows in North America, S. Cernuus species can be easily obtained in Korea, considering raw material supply and demand. Preference is given to using S. Chinensis species. In addition, this can be both natural and artificially cultivated, and all of the roots, leaves, stems, etc. of the genus Zinnia can be used for extraction.

The active ingredient having tyrosinase inhibitory activity can be extracted by using water or an organic solvent, such as methanol, acetone, propanol, ethanol, 1,3-butylene glycol, propylene glycol, alone or in a mixture of Triticala plant. .

When using a water or a mixed solvent with water, it is preferable to add 6-12 L of solvent per kg of raw materials for 300 seconds, and to extract for 6 to 12 hours in the temperature range of 60-80 degreeC.

When using an organic solvent only or a mixture as a solvent, it is preferable to add 6-10 L of solvent per kg of a 300 second raw material, and to extract for 12 to 14 hours in the temperature range of 18-23 degreeC.

After the extraction is completed, primary filtration is carried out, and it is aged for 5 to 10 days at room temperature (1 to 30 ° C). When the extract is aged at room temperature for a certain period of time, the perfume, cologne, and wine are aged for a long time to have a soft and subtle aroma and taste. Elution of the active ingredients (vitamin C, tannin, free amino acids, free sugars, etc.) is easier to elution than the extract concentrated under reduced pressure immediately, and the whitening effect is better. At this time, if left for 5 days or less, the medicinal herb smell remains and the active ingredient content is low, if more than 10 days, there is no herbal odor but the content of the active ingredient is no longer increased.

The aged extract is again filtered and concentrated to obtain a solvent completely removed. Concentration is preferably performed under reduced pressure of 25 to 35 cmHg. This is because when it is less than 25 cmHg, the concentration time is long, and when it is 35 cmHg, a large amount of active ingredient to be concentrated is discharged. The concentrated extract is a viscous liquid, sticky dark brown or dark brown. This liquid may be powdered through high temperature drying (80-100 ° C.) or lyophilization, but since there is no difference in efficacy before and after powdering, this step may be omitted to simplify the process.

The three hundred sec extract according to the present invention is an excellent tyrosinase inhibitor, and has an excellent whitening effect. Therefore, it can be added to all cosmetic compositions related to whitening, such as flexible lotion, nourishing lotion, nourishing cream, massage cream, essence and pack, the amount of which is 0.0001 to 20% by weight, in particular 0.01 to 10% by weight, based on the dry weight of the cosmetic composition. It is preferable to make it to an extent. It is difficult to obtain a whitening effect at concentrations of less than 0.0001 weight, and compounding at more than 20 weight is uneconomical in terms of increasing the whitening effect or stability of the product.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are merely illustrative of the present invention and the scope of the present invention is not limited to these examples.

Example

In the following examples, as the raw material for extraction, three hundred seconds artificially cultivated and naturally dried in Busan, roots, leaves, stems were used in the same weight.

Example 1

After extracting 500g of dry finely cut vinegar for 3 hours at 60 ℃ for 6 hours with 5L of water in an extractor equipped with a cooling capacitor, it was filtered through 400 mesh standard, left at room temperature (1 ~ 30 ℃) for 7 days, and then aged again. Filter using standard.

The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 27.5 g of a viscous trichome extract.

Example 2

After extracting 500 g of dry finely divided trichophytes in a water bath (18-23 ° C.) for 12 hours using 1.2 L of methanol, 300 ml of acetone, 300 ml of propanol, and 1.2 L of ethanol, the mixture was filtered through a 400-mesh standard and filtered at room temperature (1∼ 30 days), and aged for 7 days, and filtered again using a 400 mesh standard.

The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 39.5 g of a viscous trichome extract.

Example 3

After extracting 500 g of dry finely divided three hundred scents at 60 ° C. with 1.8 L of water and 1.2 L of 1,3-butylene glycol in an extractor equipped with a cooling condenser for 12 hours, the mixture was filtered through a 400 mesh standard and subjected to room temperature (1∼ 30 days), and aged for 7 days, and filtered again using a 400 mesh standard.

The extract was concentrated under reduced pressure using a vacuum evaporator to obtain 32.7 g of a three hundred sec extract with viscous consistency.

Example 4

After extracting 500g of dry finely cut three hundred seconds with methanol 3L in a water bath (18-23 ℃) for 12 hours, it was filtered through a 400-mesh standard, left at room temperature (1-30 ℃) for 7 days, and then aged again. Filtration was carried out using a mesh standard.

The extract was concentrated under reduced pressure using a vacuum evaporator to obtain 45.9 g of a extract of viscous trichome.

Example 5

After extracting 500 g of dry finely cut vinegar for 3 hours with 3L of ethanol in a water bath (18 ~ 23 ℃), it was filtered through 400 mesh standard, left at room temperature (1 ~ 30 ℃) for 7 days, and then aged again. Filtration was carried out using a mesh standard.

The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 40.4 g of a three hundred sec extract with viscous consistency.

Preparation Example 1

Flexible Lotion (Skin Lotion)

division Raw material name weight( ) One Purified water 83.420 2 Three hundred seconds extract of Example 1 2.000 3 Butylene glycol 5.000 4 glycerin 1.000 5 Modified ethanol 8.000 6 Polyoxyethylene Cured Castor Oil 0.300 7 Paraoxybenzoic Acid Methyl 0.100 8 Yellow No. 4 (0.2) 0.085 9 Spices (E-5177) 0.095

According to the composition, the aqueous phase (the raw materials 1 to 4) and the ethanol phase (the raw materials 5 to 7) were separated and weighed, and then dissolved separately, and the ethanol phase was mixed in the aqueous phase and mixed. Subsequently, additives (the above raw materials 8 to 9) were added and mixed. It was filtered using a membrane filter and then put into a suitable container for production.

Preparation Example 2

Nutrition lotion

division Raw material name weight () One Purified water 76.005 2 Three hundred seconds extract of Example 1 4.000 3 glycerin 5.000 4 Melchillgluses 20 1.500 5 Carboxy Vinyl Polymer 0.200 6 Paraoxybenzoic Acid Methyl 0.100 7 Liquid paraffin 10.000 8 Self-emulsifying monoglycerate glycerin 1.300 9 Stearic acid 0.700 10 Monostearine polyoxyethylene sorbitan (20E.O) 0.600 11 Cetanol 0.300 12 Triethanolamine 0.200 13 Spices (E-5177) 0.095

According to the composition, the aqueous phase (the raw materials 1 to 6) and the oil phase (the raw materials 7 to 11) were separated and weighed, and then mixed and dissolved separately at 80 ± 2 ° C., and the oil phase was mixed with the aqueous phase. Subsequently, the raw material 12, a neutralizing agent, was added thereto, neutralized, and then cooled to 50 ° C. After cooling, the raw material 13 was added and then cooled to 35 ° C. in a suitable container to produce a product.

Preparation Example 3

Nutrition Cream

division Raw material name weight () One Purified water 73.855 2 Three hundred seconds extract of Example 1 5.000 3 glycerin 4.000 4 Methylgluses 20 2.000 5 Carboxy Vinyl Polymer 0.150 6 Paraoxybenzoic Acid Methyl 0.100 7 Liquid paraffin 7.000 8 Self-emulsifying monoglycerate glycerin 1.000 9 Stearic acid 3.000 10 Monostearine polyoxyethylene sorbitan (20E.O) 1.500 11 Cetanol 2.000 12 Triethanolamine 0.300 13 Spices (E-5177) 0.095

According to the composition, it was prepared in the same manner as in Preparation Example 2.

Preparation Example 4

essence

division Raw material name weight () One Three hundred seconds extract of Example 1 5.000 2 glycerin 5.000 3 Butylene glycol 3.000 4 Spices (E-5177) 0.095 5 Rose water 86.905

According to the composition, the raw materials were measured in order, mixed and dissolved, and then filtered through a 200 mesh sieve, and put into a suitable container to produce a product.

Test Example 1

The tyrosinase inhibitory effect and arbutin tyrosinase inhibitory effect on the three hundred sec extracts obtained in Examples 1 to 5 were compared.

Enzyme tyrosinase was isolated and purified from Mushroom (Mushroom) was used Sigma (Sigma). First, L-tyrosine (manufactured by Sigma), which was a substrate, was made into a solution of 0.3 mg / ml in distilled water, and 1.0 ml of this solution was taken in a test tube.

Here, 1.0 ml of potassium phosphate buffer (Ptastium phosphate buffer: pH 6.8, 0.1M) and 0.7 ml of each step dilution solution and arbutin (hardener) of each extract of Examples 1 to 5 were added to the reaction solution, followed by 37 ° C. The reaction was carried out for 10 minutes at. Diluted solvent was ethanol and water 7: 3 or 3: 7 in a total amount of 50ml and as a control, 0.7 ml of diluting solvent was used instead of each extract.

0.1 ml each of tyrosinase (1,250 units / ml) was added to the reaction solution, and the resultant was reacted for 10 minutes in a 37 ° C thermostat. The test tube containing the reaction solution was placed in iced water and quenched to stop the reaction, and the absorbance at wavelength 475 nm was measured with an absorbance spectrometer (160-A, Shimazu).

The tyrosinase inhibitory effect of each extract was determined by the following formula.

The dose (IC ₅₀ = Inhibition Concentration 50) of each sample for reducing tyrosinase activity by 50 was determined from the tyrosinase inhibition rate of the dilution solution of each sample obtained by the above formula. The test results are shown in Table 2 below.

sample IC ₅₀ Example 1 0.039 Example 2 0.017 Example 3 0.030 Example 4 0.025 Example 5 0.021 Arbutin 0.190 IC ₅₀ (Inhibition Concentration 50): The dose of each sample that inhibits tyrosinase activity by 50 in the reaction system.

As shown in Table 2, the three hundred sec extract has very strong tyrosinase activity inhibition compared to arbutin.

Test Example 2

Inhibition test of melanin biosynthesis of B16 melanoma cells to the three hundred sec extract obtained in Example 1 and arbutin and kojic acid. Mouse-derived B16 melanoma cells (American Type Culture Collection, Accession No .: 6323) were inoculated into DMEM (Dulbecco's Modified Eagles Medium (Gibco, USA)) medium containing 4.5 g / L glucose and 10 fetal bovine serum antibiotics. Incubated at 37 ° C in a ml T-flask. After incubating for 24 hours under 5CO ₂ condition, cells were isolated by treatment with 0.05 trypsin containing 0.02EDTA, and then inoculated in 50 ml T-flask and incubated for 48 hours. At this time the cell number was 5 × 10 ⁴ cells / flask. Here, three hundred sec extract, appropriate concentration (10 μl, 50 μl), arbutin (curing agent) and kojic acid (manufactured by Sigma) were diluted in DMEM medium, treated with cultured melanoma cells, and incubated at 37 ° C. for 5 days. After incubation, the medium was removed, the cells were separated by treatment with 1 ml of phosphate buffered saline containing 0.02EDTA and 0.05 trypsin, and then centrifuged (1500 rpm) for 5 minutes to absorb the precipitated melanin at 475 nm. It measured with the photometer (160-A, Shimazu make). The tyrosinase activity inhibitory effect () was calculated | required as a ratio with respect to the melanin quantity in the case of addition of a sample.

sample Inhibition rate () Three hundred sec extract 50 μl 81.2 10 μl 56.0 Arbutin 50 μl 40.1 10 μl 24.2 Kojic acid 50 μl 75.9 10 μl 15.8

As shown in Table 3, it can be seen that the three hundred seconds extract has a much better melanin production inhibitory effect than arbutin, kojic acid.

Test Example 3

Human body application test

A pin chamber (Epitest Ltd) for 50 adult males (25 males and 25 females) with a cream sample (manufactured by the method of Preparation Example 3) containing 5 weights of three hundred sec extracts obtained in Examples 1 to 5 of the present invention. After each sample was applied to the oy agent) and attached to the inner soft skin of the upper arm, skin conditions such as irritation, erythema, itching, and allergy were observed after 24 hours, and again after 24 hours.

Number of skin condition protons / total subjects for sample number sample 24 hours later 48 hours later One Cream containing three hundred seconds extract 5 (weight) obtained in Example 1 0/50 0/50 2 Cream containing three hundred seconds extract 5 (weight) obtained in Example 2 1/50 1/50 3 Cream containing three hundred seconds extract 5 (weight) obtained in Example 3 0/50 0/50 4 Cream containing three hundred seconds extract 5 (weight) obtained in Example 4 0/50 0/50 5 Cream containing 300 seconds extract 5 (weight) obtained in Example 5 0/50 0/50

As shown in Table 4, the cream containing three weights of three hundred seconds extract is a positive reaction only one person in the three hundred seconds extract of Example 2, the rest do not show a positive reaction, it can be confirmed that the safety for human skin is excellent.

Experimental Example 4

The whitening effect of the cosmetic containing the tritical herb extract of the present invention was evaluated through a practical use test. Production Example 4 containing the extract of Triticalum extract and Production Example 4 of the same manufacturing method, except that it did not contain the Extract of Triticalum extract, were used as controls, and 20 females of 20 to 45 years old who had only spots on the face and no other skin diseases The blemish healing effect was tested.

To 20 subjects, the preparation of Example 4 containing triticale extract on the left side of the face and Preparation 4 (control group) containing no triticale extract on the right side were applied twice a morning and in the evening, followed by a proper amount for 2 months. The results were visually observed to evaluate the degree of discoloration of the spots by comparison with the control group.

As shown in Table 5, the cosmetic product of the present invention containing tritical herb extract proved to have a much better whitening effect than the control group.

Compared with the arbutin and kojic acid developed previously, the tritical herb extract according to the present invention has been shown to be very excellent in terms of safety as well as whitening effect by showing the most excellent tyrosinase inhibitory effect.

In addition, the above three hundred seconds extract can be applied to the skin whitening cosmetics and showed an excellent whitening effect when blended in cosmetics.

Claims (5)

A tyrosinase inhibitor consisting of three hundred seconds extract. Whitening cosmetics containing tritical extract. The whitening cosmetic according to claim 2, wherein the tritical herb extract is contained in an amount of 0.0001 to 20% by weight based on the dry weight of the cosmetic composition. 6 to 12 liters of a mixed solvent of water or water and at least one solvent selected from methanol, acetone, propanol, ethanol, 1,3-butylene glycol and propylene glycol is added per kg of raw material for 300 seconds, and 60 to 80 ° C. 6 to 12 hours after extraction, filtration, and the step of aging for 5-10 days at 1 to 30 ℃, comprising a method for producing a tyrosinase inhibitor according to claim 1. 6 kg of one or more solvents selected from methanol, acetone, propanol, ethanol, 1,3-butylene glycol and propylene glycol were added per kg of raw material for 300 seconds, followed by extraction and filtering at 18 to 23 ° C for 12 to 14 hours. A method for preparing a tyrosinase inhibitor according to claim 1, comprising the step of aging at 1 to 30 ° C. for 5-10 days.