JP2011001283A - Apigenin-containing composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a composition stably containing apigenin.SOLUTION: The composition contains not less than 3 ppm apigenin and dibutylhydroxytoluene and has a pH of lower than 7. A method for producing an apigenin-containing composition comprises mixing not less than 3 ppm apigenin with dibutylhydroxytoluene and adjusting the pH of the composition to lower than 7.

Description

  The present invention relates to a composition that stably retains apigenin.

Apigenin is a flavonoid contained in plants such as chamomile ( Matricaria recutita L.), Roman chamomile ( Anthemis nobilis L.), Dahlia ( Dahlia pinnata ), Fujimodoki ( Daphne genkwa ), Sorghum nervosum Bess, Urease activity inhibitory action (patent document 1), antioxidant action (patent document 2), melanin production promoting action (patent document 3, patent document 4), anti-inflammatory action (patent document 5), and pigmentation inhibitory action (patent document) 6) is useful as a component of cosmetics, pharmaceuticals and quasi drugs. Therefore, various compositions containing apigenin or an apigenin-containing plant extract have been reported (Patent Documents 2 to 6).

JP 2004-91338 A JP 2005-289880 A JP-A-9-263534 JP 2004-2264 A JP 2007-8847 A JP 2006-327988 A

  In the process of studying a composition containing apigenin at a high concentration, the present inventors coexisted with a composition containing apigenin, dibutylhydroxytoluene (BHT), which is a substance generally used as an antioxidant. In this case, it was found that there is a problem that apigenin and BHT form an adduct, and as a result, apigenin cannot be stably maintained in the composition.

  Therefore, the present inventors have studied to maintain apigenin stably in the composition in the presence of BHT. As a result, it was found that apigenin can be stably maintained in the composition by adjusting the pH of the composition to less than 7.

That is, the present invention relates to a composition comprising 3 ppm or more of apigenin and dibutylhydroxytoluene and having a pH of less than 7.
The present invention also relates to a method for producing an apigenin-containing composition characterized by blending 3 ppm or more of apigenin and dibutylhydroxytoluene and adjusting the pH of the composition to less than 7.
The present invention also relates to a method for stabilizing apigenin, comprising adjusting the pH of the composition to less than 7 in a composition containing apigenin and dibutylhydroxytoluene.

  According to the present invention, it is possible to provide a composition that stably contains apigenin, and to provide a pharmaceutical, quasi-drug, or cosmetic that can maximize the effects of apigenin as an active ingredient. it can.

In the present invention, apigenin means a compound represented by the following formula.

Apigenin can be obtained by methods commonly known in the art. Typically, it can be obtained from a plant containing apigenin, for example, a plant such as chamomile, roman chamomile, dahlia, Fujimodoki, Koryo or an extract thereof, preferably a high concentration plant extract. Here, chamomile means Matricaria recutita L. of the asteraceae family, and is also called German chamomile or German chamomile. Roman chamomile means Anthemis nobilis L., Asteraceae, and Dahlia means Dahlia pinnata , Asteraceae. Moreover, Fujimodoki means Daphne genkwa of the Chrysomelidae family, and Koryo means Sorghum nervosum Bess of the Gramineae family.
Among the above plants, Asteraceae plants are preferable, and chamomile and roman chamomile are more preferable, and roman chamomile is more preferable in that a high concentration apigenin-containing extract is obtained.

  Apigenin-containing plants include any part of the plants listed above, for example, whole grass, leaves, stems, buds, flowers, buds, wood parts, bark, lichens, roots, rhizomes, corms, corms, tubers, Seeds, fruits, mycorrhiza, resin, etc., or combinations thereof can be used. In the case of chamomile and roman chamomile, it is preferred to use a corolla or a bud. These parts are added to the composition of the present invention as they are, after being pulverized, cut or dried, or further through a process such as extraction.

  Examples of plant extracts containing apigenin include various solvent extracts obtained by conventional methods, or diluted solutions thereof, concentrated solutions thereof, dried powders thereof, or activated carbon treatments thereof. Of these, various solvent extracts are preferred. Or the extract of the said plant marketed can also be used. As a specific example of the extraction method, a plant is immersed in an extraction solvent 5 to 40 times (mass ratio) of the extraction raw material, extracted at room temperature or under reflux heating for 1 day to 1 month, and then filtered to remove the residue. The method of doing is mentioned.

  As the extraction solvent, either a polar solvent or a nonpolar solvent can be used. Specific examples of the extraction solvent include water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; methyl acetate and ethyl acetate. Esters such as tetrahydrofuran; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as squalane, hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as toluene; dichloromethane , Halogenated hydrocarbons such as chloroform and dichloroethane; and carbon dioxide. These can be used as a mixture. Among these, preferred specific examples include an aqueous ethanol solution, an aqueous butanediol solution, and an aqueous propylene glycol solution, more preferably an aqueous ethanol solution and an aqueous butanediol solution, and even more preferably an aqueous ethanol solution. As the said ethanol aqueous solution, 5-99.5 volume% ethanol aqueous solution is preferable, and 10-80 volume% ethanol aqueous solution is more preferable. When using chamomile, a 5-55% by volume ethanol aqueous solution is preferred, and a 10-50% by volume ethanol aqueous solution is more preferred. When using Roman chamomile, a 30 to 99.5% by volume ethanol aqueous solution is preferable, and a 40 to 80% by volume ethanol aqueous solution is more preferable. The amount of the solvent used is about 10 to 40 times (mass ratio). The extraction time is preferably 1 day to 1 month, and the extraction temperature is preferably 5 to 80 ° C.

  More preferably, according to the description in Japanese Patent Application No. 2009-004378, the extract obtained as described above is further subjected to the following three steps of adsorption step, washing step and elution step to extract containing higher concentrations of apigenin. You can get things.

  In the adsorption step, for example, the adsorbate is obtained by bringing the extract and the adsorption carrier into contact or in contact with each other. Preferable specific examples of the adsorption carrier include powdered cellulose, radiolite, celite and the like, preferably powdered cellulose, radiolite and combinations thereof. The adsorption carrier is preferably used in an amount of about 2.5 to 10 g per 200 mL of the extract from the viewpoint of the recovery rate of apigenin. In order to enhance the adsorption of apigenin to the adsorption carrier, it is preferable to concentrate the extract of the apigenin-containing plant after the contact or the like. The concentration may be performed according to a conventional method, and specific examples include concentration under reduced pressure.

The washing step is performed, for example, by bringing the adsorbate obtained in the previous step into contact with 2 to 10 times the volume of water or 20% by volume or less of an aqueous ethanol solution. Contact with water is preferred.
Specific means of washing include, for example, filtration and decantation, preferably atmospheric filtration using filter paper or filter cloth; filter press, pressurized filter paper filter, leaf filter, rotary press, etc. Pressure filtration; vacuum filtration using a rotary drum type continuous filter, vacuum filter, and the like. Normal pressure filtration is preferable in terms of simplicity. When washing by filtration, apigenin is retained in the adsorption carrier and the others are separated as a filtrate.

  The elution step is performed by bringing the washed adsorbate into contact with 2 to 10 times volume of 40 to 99.5% by volume ethanol aqueous solution. Specific examples of elution include the same means as in the above washing. In the case of elution by filtration, apigenin is separated from the adsorption carrier as a filtrate.

  By applying the above three steps, a high-concentration apigenin-containing extract containing 10-30% by mass, preferably 15-25% by mass, of apigenin per evaporation residue is obtained.

  The obtained high-concentration apigenin-containing extract is further subjected to chromatography, liquid-liquid partition, and the like, whereby inactive impurities in the extract can be removed.

  The high-concentration apigenin-containing extract can be used as it is, but it may be used after being diluted, concentrated, or lyophilized, and then prepared into a powder or paste.

  Dibutylhydroxytoluene (BHT) is a well-known substance used as an antioxidant and the like, and a commercially available product (for example, Wako Pure Chemical Industries, Ltd., Code No. 047-29451) can be purchased.

  The composition of the present invention can be produced by blending apigenin and BHT and adjusting the pH of the composition to less than 7. Thereby, adduct formation between BHT and apigenin in the composition is suppressed, and apigenin is stably maintained in the composition.

  The pH of the composition of the present invention is not particularly limited as long as it is adjusted to less than 7, and may be appropriately adjusted according to the use of the composition. The pH value of the composition is preferably less than 7, more preferably 6.8 or less, still more preferably 6.5 or less, and even more preferably within the range of pH 4 to 6.5. The pH of the composition can be adjusted with a pH adjuster. The pH adjuster is not particularly limited as long as it is usually used. For example, inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid; citric acid, acetic acid, lactic acid, succinic acid, glycolic acid, ascorbine Organic acids such as acid, malic acid, fumaric acid, tartaric acid, urea, ε-aminocaproic acid and pyrrolidone carboxylic acid; betaines such as glycine betaine and lysine betaine; inorganic alkalis such as metal hydroxide; guanidine, 2-amino-2 -Organic amines such as methylpropane; alkanolamines such as ammonia, monoethanolamine, diethanolamine, triethanolamine, isopropanolamine, diisopropanolamine; basic amino acids such as arginine and lysine, and combinations thereof.

  In the present invention, when apigenin is stably maintained in the composition, the contents (absolute amounts) of apigenin and BHT in the composition are not particularly limited, and may be appropriately adjusted according to the dosage form, use, etc. of the composition. That's fine. For example, the content of apigenin may be 1 ppm by mass or more with respect to the total mass of the composition, preferably 3 ppm by mass or more, more preferably 5 to 50 ppm by mass, and even more preferably 7 to 25 ppm by mass. is there. The BHT content may be, for example, in the range of 0.001 to 500 ppm by mass, preferably 0.01 to 10 ppm by mass with respect to the total mass of the composition. When the apigenin-containing extract is used in the production of the composition of the present invention, an appropriate amount of the extract is blended with the composition of the present invention so that the apigenin content in the final composition falls within the above range.

  In addition to the above-mentioned apigenin, BHT and pH adjuster, the composition of the present invention may be added with other active ingredients or arbitrary additives in an appropriate amount in accordance with the dosage form, application, etc. of the composition. it can. Examples of additives include excipients, binders, disintegrants, lubricants, diluents, osmotic pressure regulators, emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet absorbers, and humectants. , Thickeners, brighteners, activity enhancers, anti-inflammatory agents, bactericides, fragrances, flavoring agents, flavoring agents and the like. As other active ingredients, for example, in the case of cosmetic compositions, active ingredients other than apigenin, such as moisturizers, whitening agents, UV protection agents, cell activators, detergents, keratolytic agents, makeup ingredients (for example, Other cosmetic / cosmetic ingredients such as makeup base, foundation, funny, powder, teak, lipstick, eye makeup, eyebrow, mascara, etc. may be contained.

  The composition of the present invention can be used as pharmaceuticals, quasi drugs, cosmetics, etc., or as raw materials thereof. The form of the composition is arbitrarily selected according to its use. For example, in the case of pharmaceuticals and quasi-drugs, tablets, pills, capsules, solutions, syrups, powders, granules, etc. for oral administration, injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation And forms for parenteral administration such as solutions, emulsions, suspensions and the like for suppositories and boluses, and forms for topical administration such as creams, lotions, gels, sheets, patches, sticks and the like. As a form when using a composition as cosmetics, the arbitrary forms which can be used for cosmetics, such as cream, emulsion, lotion, suspension, gel, powder, pack, sheet, patch, stick, cake, etc., are mentioned. .

  According to the present invention, since the adduct formation between BHT and apigenin contained in the composition is suppressed, apigenin in the composition is stably maintained. Therefore, according to the present invention, it is possible to provide a composition that stably contains apigenin as an active ingredient and, as a result, maximizes the effect of apigenin. In the composition provided by the present invention, the action of apigenin (for example, urease activity inhibitory action, antioxidant action, melanin production promoting action, anti-inflammatory action, pigmentation inhibiting action, or whitening action) is not reduced. As a result, the composition exhibits an excellent effect.

  The present invention will be described in more detail based on the following reference examples and examples, but the present invention is not limited to these contents. In the following, ppm represents ppm by mass.

Example 1 Stability Evaluation of Apigenin in Composition 20 mg of Roman chamomile was extracted with 200 mL of 50% butanediol at room temperature for 7 days, followed by filtration to obtain 155 mL of Roman chamomile extract. The evaporation residue per mL of the extract was 18.6 mg, and the amount of apigenin was 305 ppm. This extract was used as an apigenin-containing extract in subsequent experiments.

  Solutions having different pH conditions were prepared according to the formulation shown in Table 1 below. BHT was added to each pH condition solution at 0, 1, 10, 100, and 500 ppm to prepare a test solution (apigenin final concentration of about 9 ppm). After each test solution was stored at −5 ° C. and 50 ° C. for 1 month, the amount of apigenin in the solution was determined by HPLC (Inert Sil ODS3 (manufactured by GL Sciences Inc., inner diameter 3.0 mm, length 150 mm, particle diameter 5 μm)). Eluent 50% 25 mM aqueous potassium dihydrogen phosphate / 50% methanol; column temperature 40 degrees; detection UV 340 nm). The ratio of the amount of apigenin at 50 ° C. to the amount of apigenin at −5 ° C. was determined as the apigenin residual rate.

  The results are shown in Table 2 below. When the mass ratio of apigenin to BHT was less than about 1 (BHT 10 ppm or higher) and pH 7 or higher, the apigenin residual ratio was decreased, indicating that the adduct formation between apigenin and BHT proceeded. On the other hand, when the mass ratio of apigenin to BHT was about 1 or more, or when the pH of the solution was less than 7, adduct formation did not occur, and a high residual rate was maintained even after long-term storage. That is, it was shown that apigenin can be favorably retained by adjusting the pH of the solution to less than 7.

  In the test solution having a reduced apigenin residual rate, an adduct of apigenin and BHT of the following formula was formed.

Reference example Whitening effect of apigenin (preparation of test sample)
Using the Roman chamomile extract prepared in Example 1, test samples having the compositions shown in Table 4 were prepared, and the whitening effect of apigenin was evaluated.

(Whitening effect evaluation)
The skin color of the inner side of the upper arm of 10 healthy male and female subjects was measured with a color difference meter (CMS-35FS, manufactured by Murakami Color Research Co., Ltd.), and the initial L ^* value was calculated from the obtained Munsell value. The skin at the site where the skin color was measured was irradiated once with UV rays in the UV-B region, which was twice the minimum erythema amount (2 MED), using a Toshiba FS-20SE lamp. Then, the sample (Test 1, 2, Comparative product 1) was continuously applied to the same site twice a day for 3 weeks. The skin color was measured again with a color difference meter, and the L ^* value was calculated from the obtained Munsell value. The amount of change (ΔL ^* ) with respect to the initial value of the L ^* value of the skin of the test product 1 and 2 and the comparative product 1 applied part was calculated. For each sample, the average value of ΔL ^* values (n = 10) was calculated. Seeking risk rate (p value) by performing a paired t-test with a [Delta] L ^* value the average value of the comparative product 1 coated portion of skin and [Delta] L ^* value the average value of the test sample 1 and 2 coating unit skin whitening evaluation It was used as an index. The results are shown in Table 5.

From the results in Table 2, the 5% by weight Roman chamomile extract blend (containing 15 ppm apigenin) has a significant ΔL ^* value inhibitory effect compared to the comparative product (placebo) and is excellent in whitening effect. It has been shown. Even in the case of the 1% by mass Roman chamomile extract blend (containing 3 ppm apigenin), a tendency to suppress the ΔL ^* value was recognized as compared with the comparative product (placebo), and the whitening effect was estimated.

Preparation Example A whitening composition A pH 6 lotion having the following composition was prepared.
(Composition) (Composition: Mass%)
Roman chamomile extract 5
Ethanol 20
1,3-Butylene Glycol 25
BHT 0.001
Purified water balance

Claims (5)

  A composition comprising 3 ppm or more of apigenin and dibutylhydroxytoluene and having a pH of less than 7.   The composition according to claim 1, which is a whitening composition.   A method for producing an apigenin-containing composition comprising blending 3 ppm or more of apigenin and dibutylhydroxytoluene, and adjusting the pH of the composition to less than 7.   The method according to claim 3, wherein the composition is a whitening composition.   A method for stabilizing apigenin, comprising adjusting the pH of the composition to less than 7 in a composition containing apigenin and dibutylhydroxytoluene.