JP2005089376A - Bleaching agent and antioxidant - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a bleaching agent and an antioxidant by discovering an active ingredient having an excellent bleaching action and an antioxidant action. <P>SOLUTION: An extract of a plant belonging to the genus Tribulus having the excellent bleaching action and antioxidant action is used as the bleaching agent and the antioxidant. The extract of Tribulus terrestris is preferably used as the extract of the plant belonging to the genus Tribulus. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

The present invention relates to a whitening agent and an antioxidant. More specifically, the present invention relates to a whitening agent and an antioxidant containing as an active ingredient an extract of one or more kinds of plants selected from the plant of the genus Tribulus .

  Pigmentation symptoms caused by spots, sunbacus and sunburn are thought to be caused by ultraviolet rays, female hormones, genetic factors, etc., but the factors vary from person to person. Therefore, as a method for improving pigmentation symptoms, a method for suppressing the production of melanin that causes pigmentation, a method for suppressing the activity of the enzyme tyrosinase, a method for suppressing oxidation by ultraviolet rays using an antioxidant, and the like have been studied. Various whitening agents and antioxidants have been reported so far. As whitening agents reported so far, hinokitiol derivatives (see Patent Document 1), ergosterol derivatives (see Patent Document 2), and phenoxyacetic acid derivatives (see Patent Document 3) are known as antioxidants. As an example, an extract of a plant belonging to the genus Sarogase is known (see Patent Document 4). However, the whitening agents and antioxidants that have been reported so far are not always effective, are restricted in use due to safety issues, and give undesirable color and odor to the preparation. At present, sufficient products have not yet been obtained in terms of effects, side effects, and stability.

  In addition, the prior art regarding the whitening agent and antioxidant which use the extract of Trivulus genus plant as an active ingredient was not recognized.

JP 2002-047167 A Japanese Patent Application Laid-Open No. 2002-114685 JP 2001-354511 A Japanese Patent Laid-Open No. 10-182413

  In modern times, people's interest in whitening to improve pigmentation symptoms is very high, and development of better whitening agents and antioxidants is expected. It was made. Accordingly, an object of the present invention is to provide a whitening agent and an antioxidant that exhibit excellent effects.

In order to find a whitening agent and an antioxidant that exhibit excellent effects, the present inventors have examined various whitening effects and antioxidative effects. As a result, it has been found that an extract of one or more kinds of plants selected from the genus Tribulus exhibits an excellent whitening effect and antioxidant effect, and further studies are made to complete the present invention. It came.

That is, the present invention relates to a whitening agent and an antioxidant containing, as active ingredients, one or two or more plant extracts selected from the genus Tribulus . As an extract of a plant selected from plants belonging to the genus Tribulus, an extract of Tribulus terrestris can be preferably used.

  According to the present invention, it is possible to provide a whitening agent and an antioxidant having an excellent effect, and by using the whitening agent, it is excellent in preventing and improving pigmentation symptoms caused by spots, freckles, ultraviolet rays and the like. Show the effect.

The plant used in the present invention is not particularly limited as long as it is a plant belonging to the genus Tribulus . About 20 species of genus Tribulus are known, but due to their relative availability and effectiveness, Tribulus terrestris , Tribulus cistoides , Tribulus maximus ( Tribulus maximus ) or the like can be preferably used, and in particular, Tribulus telestris is preferable from the viewpoint of effectiveness.

  When using these plant species, it is common to use an extract. For extraction, any part of Tribulus plant stem, branch, fruit, leaf, flower, seed, bark, sap, root, bud, etc. may be used, but it is effective to use seed. And preferred from the viewpoint of simplicity.

  In the extraction, it may be used as it is, but considering the extraction efficiency, it is preferable to perform the extraction after performing processing such as shredding, drying, and pulverization. The extraction method is not particularly limited as long as it is a conventional method, and examples include a method of immersing in an extraction solvent and an extraction method using a supercritical fluid or a subcritical fluid. In order to increase the extraction efficiency, stirring or homogenization can be performed in the extraction solvent. Although extraction temperature is not specifically limited, It is appropriate to set it as the temperature below the boiling point of an extraction solvent from about 5 degreeC. Since the extraction time varies depending on the type of extraction solvent and the extraction temperature, it is not particularly limited, but is appropriately about 1 hour to 14 days.

  The extraction solvent is not particularly limited, but water, alcohols (ethanol, methanol, isopropanol, isobutanol, n-hexanol, methyl amyl alcohol, 2-ethylbutanol, n-octyl alcohol, etc., monovalent alcohol, glycerin, ethylene glycol , Ethylene glycol monomethyl ether, propylene glycol, propylene glycol monomethyl ether, propylene glycol monoethyl ether, polyethylene alcohol such as triethylene glycol, 1,3-butylene glycol, hexylene glycol or derivatives thereof, ketones (acetone, methyl ethyl ketone) , Methyl isobutyl ketone, methyl-n-propyl ketone, etc.), esters (ethyl acetate, isopropyl acetate, etc.), ethers (ethyl ether, Sopropyl ether, n-butyl ether, etc.), polar solvents such as acetonitrile, liquid hydrocarbons such as n-pentane, n-hexane, non- (low) polar solvents such as supercritical fluid of carbon dioxide, physiological saline, phosphoric acid A buffer solution, a phosphate buffered saline, etc. can be mentioned, These can be used individually or in mixture of 2 or more types. Examples of the solvent include water and alcohols (ethanol, methanol, isopropanol, isobutanol, n-hexanol, methyl amyl alcohol, 2-ethylbutanol, n-octyl alcohol and the like monovalent alcohols, glycerin, ethylene glycol, ethylene glycol). Use one or more of monomethyl ether, propylene glycol, propylene glycol monomethyl ether, propylene glycol monoethyl ether, triethylene glycol, 1,3-butylene glycol, hexylene glycol and the like) Is preferable from the viewpoint of ease of extraction and effectiveness of the extract, and it is particularly preferable to use a mixture of water and ethanol.

  The extract of Tribulus genus plant can be used as it is, but the concentrated and dried product can be redissolved in water or a polar solvent, or decolorized and deodorized as long as these physiological functions are not impaired. It may be used after a purification treatment such as desalting or a fractionation treatment by column chromatography or the like. The said extract of Trivulus genus plant, its processed material, and a fractionated product can also be used after freeze-drying after each processing and fractionation, and melt | dissolving in a solvent at the time of use.

  A whitening agent containing an extract of Tribulus spp. As an active ingredient has excellent tyrosinase activity inhibitory action and melanin production inhibitory action, and is excellent in preventing and improving various pigmentation symptoms such as spots and freckles Demonstrate the effect.

  An antioxidant comprising an extract of the genus Tribulus as an active ingredient has an excellent radical scavenging action and exhibits an excellent effect in preventing and improving various symptoms caused by oxidative damage.

  In addition, whitening agents and antioxidants containing Tribulus plant extracts as active ingredients can be used alone, but as whitening ingredients or antioxidant ingredients, such as pharmaceuticals, quasi drugs, foods and drinks, cosmetics, etc. By blending with various compositions, a composition having a whitening effect and an antioxidant effect can be obtained. The resulting composition having a whitening effect and an antioxidant effect is effective in preventing and improving pigmentation symptoms such as spots and freckles caused by sunburn and the like and symptoms caused by oxidative disorders.

  The dosage form of the composition containing the whitening agent and antioxidant containing the extract of Tribulus genus plant as an active ingredient is arbitrary, but if the composition is an external preparation for skin, a cleaning agent, a bath preparation, etc., it is a lotion. Can be provided as various dosage forms such as a solubilization system such as emulsion, an emulsification system such as emulsion, a dispersion system such as calamine lotion, an aerosol filled with a propellant, an ointment, a powder, and a granule. In addition, when the composition is an oral drug or food and drink, it should be a general dosage form such as liquid preparations such as drinks and drops, solid preparations such as gums and candy, capsules, powders, granules and tablets. be able to.

  In addition to these, a composition containing a whitening agent and an antioxidant containing an extract of the plant of the genus Tribrus as an active ingredient is usually a pharmaceutical, a quasi-drug, a skin cosmetic, a bath preparation, Detergents, oily ingredients blended in foods, moisturizers, powders, pigments, emulsifiers, solubilizers, detergents, UV absorbers, thickeners, drugs, fragrances, resins, alcohols, nutrient enhancement substances, A seasoning etc. can be mix | blended suitably and also combined use with another whitening agent and antioxidant is also possible.

  The amount of whitening agent and antioxidant containing one or more plant extracts selected from Trivulus plants as active ingredients should be adjusted according to the type and purpose of the composition. However, from the viewpoint of effectiveness and usability, it is preferably 0.0001 to 75% by weight, more preferably 0.001 to 50% by weight, and most preferably 0.01% to the total amount of the composition. ~ 25 weight.

  Below, the manufacture example of the extract of Trivulus genus plant and the test for evaluating each effect | action are demonstrated in detail, However, The technical scope of this invention is not limited at all by this.

[Production Example 1]
An 80-fold amount of 50% by weight ethanol aqueous solution was added to the dried pulverized seeds of the genus Tribrus and extracted for 4 hours while stirring at room temperature. The extract was collected by filtration, and after removing the solvent, a Tribulus plant extract was obtained.

[Production Example 2]
Ten times the amount of water was added to the dried ground pulverized seeds of the genus Tribrus, and the mixture was extracted by refluxing at 90 ° C. for 2 hours. The extract was collected by filtration, and after removing the solvent, a Tribulus plant extract was obtained.

[Production Example 3]
Ten times the amount of methanol was added to the dried ground pulverized seeds of the genus Tribrus and soaked at room temperature for 7 days. The extract was collected by filtration, and after removing the solvent, a Tribulus plant extract was obtained.

[Production Example 4]
The seeds of the genus Tribulus were put into a supercritical extraction apparatus, and extracted using a supercritical fluid of carbon dioxide at 40 ° C. under a pressure of 15 MPa. The extract was collected to obtain a Tribulus plant extract.

[Evaluation of melanin production inhibitory effect]
Next, it shows about evaluation of the melanin production inhibitory effect of Tribulus genus plant extract. As a sample, Tribulus terrestris extract extracted from Tribulus terrestris using Production Example 1 was used.

  The evaluation was performed according to the following procedure. B16 mouse melanoma F0 strain (B16F0) cells were seeded in a 35 mm dish at 2000 cells per dish. After culturing for 24 hours, the culture medium was replaced with Dulbecco's modified Eagle medium (DMEM) supplemented with 5 wt% fetal calf serum (FCS) to which a sample of an arbitrary concentration was added. After culturing for 7 days, the cells were collected using 0.25% trypsin, transferred to a 1.5 mL microtube, and centrifuged to obtain a cell precipitate. Furthermore, instead of the medium to which the sample was added, Dulbecco's modified Eagle medium (DMEM) to which 5 wt% fetal calf serum (FCS) was added was used as a blank, and visual determination was similarly performed. Moreover, the visual judgment was performed by five-step evaluation as shown in Table 1. Table 2 shows the determination results.

  As is clear from Table 2, when a medium supplemented with Tribulus terrestris extract was used, a significant melanin production inhibitory effect was observed. In particular, no blackening was observed when 0.25 mg / mL of Tribulus terrestris extract was added. From this, it was revealed that Tribulus terrestris extract has a high whitening action based on a melanin production inhibitory action.

[Evaluation of Tyrosinase Activity Inhibitory Action]
Next, evaluation of the tyrosinase activity inhibitory action of Tribulus genus plant extracts will be described. As a sample, Tribulus terrestris extract extracted from Tribulus terrestris using Production Example 1 was used.

The evaluation was performed according to the following procedure. Normal human epidermal melanocytes were seeded in a 96-well microplate so as to be 3.0 × 10 ⁴ cells per well. As a seeding medium, Medium154S manufactured by Kurabo Industries Co., Ltd. was used. After 24 hours, the medium was replaced with a medium supplemented with a sample prepared to an arbitrary concentration, and further cultured for 48 hours. Next, the cells were completely dissolved in 75 μL of 1 wt% Triton-X-containing phosphate buffer, and 50 μL thereof was used as a crude enzyme solution. To the crude enzyme solution, 50 μL of a 0.05 wt% L-dopa-containing phosphate buffer as a substrate was added and allowed to stand at 37 ° C. for 2 hours. The absorbance at 405 nm was measured immediately after the addition of the substrate and at the end of the reaction with a microplate reader, and the amount of produced dopamelanin was measured. At the same time, the amount of protein in each well was also measured to determine the amount of dopamelanin produced per unit protein. Relative values were obtained when the value of the blank with no sample added was taken as 100. The evaluation results are shown in Table 3 as relative values with the dopamelanin production amount in the blank with no sample added as 100. In the table, * and ** indicate a significance probability of less than 5% (P <0.05) with respect to the significance probability P value in the t test, and a significance probability of less than 1% (P <0.01). It is represented by **.

  As is clear from Table 3, a significant tyrosinase activity inhibitory action was observed when a medium supplemented with Tribulus terrestris extract was used. In particular, when 0.5 mg / mL of Tribulus terrestris extract was added, a significant tyrosinase activity inhibitory action was observed at a risk rate of less than 1%. From this, it was revealed that Tribulus terrestris extract has a high whitening action based on the tyrosinase activity inhibitory action.

[Evaluation of antioxidant activity]
Next, it shows about the antioxidant effect of Trivulus genus plant extract. As a sample, Tribulus terrestris extract extracted from Tribulus terrestris using Production Example 1 was used.

The evaluation was performed according to the following procedure. 100 μL of a sample solution diluted to an arbitrary concentration with a 50 wt% aqueous ethanol solution was added to a 96-well microplate. Next, 100 μL of a 1,1-diphenyl-2-picrylhydrazyl (DPPH) solution prepared with ethanol to a concentration of 0.2 mM was added to a 96-well microplate. The mixture was left in the dark with stirring, and the absorbance at 516 nm was measured after 24 hours. When the absorbance of the blank with no sample added was (A) and the absorbance when the sample was added was (B), the value of formula (1) was defined as the radical elimination rate. The evaluation results are shown in Table 4.
Formula (1) {1- (B) / (A)} × 100 (%)

As is apparent from Table 4, the Tribulus terrestris extract was found to have an excellent antioxidant effect. From this, it was revealed that Tribulus terrestris extract has a high antioxidant action based on radical scavenging action.

Claims (4)

A whitening agent comprising, as an active ingredient, an extract of one or more kinds of plants selected from the genus Tribulus . The whitening agent according to claim 1, wherein the Tribulus genus ( Tribulus ) plant is Tribulus terrestris . The antioxidant which uses the extract of the 1 type (s) or 2 or more types of plant chosen from a Tribulus genus ( Tribulus ) plant as an active ingredient. The antioxidant of Claim 3 whose Tribulus ( Tribulus ) plant is Tribulus terrestris ( Tribulus terrestris ).