JP2011219365A - Hyaluronidase inhibitor containing brown sugar extract and extract from red bean seed coat - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a hyaluronidase inhibitor and a cosmetic composition capable of improving and preventing wrinkle, flabbiness and inflammation, which are highly safe, and inexpensive to the containable degree in cosmetics used every day, whereas it is found that, when a brown sugar extract extracted from brown sugar and a red bean seed coat extract extracted from a red bean seed coat are mixed together, hyaluronidase inhibitory action is remarkably heightened.SOLUTION: The hyaluronidase inhibitor and the cosmetic composition contain the brown sugar extract and the extract from red bean (Vigna angularis) seed coat.

Description

  The present invention relates to a hyaluronidase inhibitor and cosmetic composition containing brown sugar extract and red bean (Vigna angularis) seed coat extract, the purpose of which is to moisturize the skin, improve and prevent wrinkles and sagging, and inflammation of the skin It is intended to provide a hyaluronidase inhibitor and a cosmetic composition excellent in safety.

Typical changes in human skin aging include wrinkles and sagging. The formation of wrinkles and sagging is considered to be caused by a decrease in the elasticity of the skin, and one of the factors is a decrease in hyaluronic acid due to aging, drying, oxidation, ultraviolet rays and the like.
Hyaluronic acid is a linear high-molecular polysaccharide in which disaccharide units of N-acetylglucosamine and glucuronic acid are linked, and is said to have a molecular weight of 1 million or more. In vivo, it exists in a wide range of extracellular matrix such as joints, glass bodies, skin, and brain.
Hyaluronic acid in the skin retains the lubricity and flexibility of the skin and prevents bacterial infection by retaining water in the cell gap and forming cells in a jelly-like matrix in the tissue. It is considered functional.
Therefore, when hyaluronidase activity, which is a hyaluronic acid-degrading enzyme, increases with aging or the like, the hyaluronic acid is reduced and the water retention of skin cells is lost, so that flexibility may be lost and wrinkles and sagging may be caused.

On the other hand, hyaluronidase is widely distributed in the living body and decomposes hyaluronic acid and is also known as a pro-inflammatory enzyme because it has been confirmed to have enhanced activity at the site of inflammation.
Therefore, the inhibitor of hyaluronidase can prevent the hyaluronic acid from being excessively decomposed, thereby suppressing the decrease in hyaluronic acid, improving and preventing wrinkles and sagging associated with aging, and also in terms of suppressing inflammation. Expected.

Until now, glycyrrhizic acid, cromoglycate sodium, baicalin, indomethacin, aspirin and the like have been confirmed to have high hyaluronidase activity inhibitory activity, and these are currently used as anti-inflammatory agents. However, glycyrrhizic acid and baicalin have limited usage and concentration, and sodium cromoglycate requires strict usage precautions for pregnant women. Furthermore, indomethacin and aspirin each have problems such as local side effects such as rash and itch have been observed.
In order to overcome such problems, a search for a substance having hyaluronidase inhibitory activity from plants has also been carried out. Acercia catechu (patent document 1) and leguminous plant Acacia catechu (patent document 1) It has been reported that extracts of herbal medicines (Peg Asen Yaku), Burbizane lebbeck, Uraria picta, Bauchinia forticata, and Cajanus indicus (Patent Document 2) have a hyaluronidase inhibitory action.
However, these are made from expensive herbal medicines or difficult-to-obtain plants. Hyaluronidase inhibitors and cosmetic compositions used in daily cosmetics are extracted from these plants. Has the problem of becoming too expensive.

Japanese Patent Laid-Open No. 10-053532 Japanese Unexamined Patent Publication No. 7-010768

  The present invention has been made to solve the above-mentioned problems, from brown sugar extract extracted from brown sugar produced from sugarcane that is inexpensive and readily available, and red bean seed coat currently processed as waste. It is found that mixing the extracted red bean seed coat extract significantly increases the hyaluronidase inhibitory effect, and it is inexpensive and safe enough to be contained in daily cosmetics, improving and preventing wrinkles, sagging and inflammation. It is an object of the present invention to provide a hyaluronidase inhibitor and a cosmetic composition that can be used.

The invention according to claim 1 relates to a hyaluronidase inhibitor characterized by containing brown sugar extract and red bean (Vigna angularis) seed coat extract.
The invention according to claim 2 relates to a cosmetic composition comprising the hyaluronidase activity inhibitor according to claim 1.
The invention according to claim 3 further relates to the cosmetic composition according to claim 2, further comprising at least one of a whitening agent, an active oxygen removing agent, an antioxidant and an ultraviolet ray preventing agent.

According to the inventions according to claims 1 and 2, since the mixture of brown sugar extract and red bean (Vigna angularis) seed coat extract inhibits hyaluronidase, the reduction of hyaluronic acid is suppressed and the skin is kept moist and wrinkled. It is possible to provide a hyaluronidase inhibitor and a cosmetic composition that improve and prevent sagging and suppress inflammation.
According to the invention of claim 3, wrinkles and sagging are obtained by including one or more of a whitening agent, an active oxygen remover, an antioxidant and an ultraviolet light inhibitor in addition to the brown sugar extract and the red bean seed coat extract. While improving and preventing, it can be set as the cosmetic composition which can keep skin white, or can prevent the skin damage by active oxygen, oxidation, and ultraviolet rays.

It is a graph showing the hyaluronidase inhibition rate of the mixture of brown sugar extract and red bean seed coat extract. It is a graph showing the hyaluronidase inhibition rate of the red beans seed coat extract.

  The present invention comprises a brown sugar extract extracted from brown sugar produced from sugarcane, which is an inexpensive and readily available plant, and an extract from red bean seed coat treated as waste, and inhibits hyaluronidase. Hyaluronic acid inhibitor that suppresses the degradation of hyaluronic acid, improves or prevents wrinkles and sagging, suppresses inflammation, and is excellent in safety to the skin, and a cosmetic composition containing the hyaluronidase inhibitor It is a thing.

The extraction method of brown sugar extract is demonstrated.
Sugarcane is a crop that is used as a raw material for sugar (sucrose) along with sugar beet with the scientific name Saccharum officinarum. The origin of cultivated species is New Guinea Island and its nearby islands, and it is a plant that is widely cultivated in tropical and subtropical regions around the world.
Prepare and prepare raw sugar (raw sucrose) as raw material from sugarcane, dissolve this crude sugar in an appropriate amount of water, bring it into contact with an adsorbent, adsorb the pigment component, and wash it thoroughly with water. After removal, the adsorbed dye component is extracted and separated by eluting with a solvent.

  Usually, this operation is carried out using an adsorbent column with water or solvent flowing down. The adsorbent is not limited as long as it is a nonpolar polystyrene-based adsorbent resin. For example, Amberlite XAD-1, Amberlite XAD-2 (trade name, manufactured by Rohm and Haas) and Selvachrome XAD-2 (trade name) (Manufactured by Selva) is preferably used. Selvachrome XAD-2 is particularly preferred from the viewpoint of yield. Further, the amount of the adsorbent used is 30 to 300 times (weight ratio), more preferably 50 to 200 times (weight ratio) of the amount of the contained pigment component.

  When the adsorbed dye component is eluted, it is preferable to remove the sugar sufficiently until the sweetness of the washing solution is completely eliminated by washing with water before elution. This is because the sugar remains and the amount of the sugar that can be blended is limited.

The elution of the dye component is not limited as long as it is a lower alcohol having a concentration of 20% by weight or more. Examples of the lower alcohol include methanol or ethanol, propyl alcohol, isopropyl alcohol, butanol, isobutanol, etc., but methanol or ethanol is preferable. This is because it is safe to use in cosmetics and the solvent is easily removed after extraction.
Actually, it is preferable to first elute with 20 to 30% by weight of lower alcohol, and after further elution of the color of the flowing-down solution, it is preferable to further elute with 95 to 99% by weight of high-concentration lower alcohol. The reason for this is that elution with only a low-concentration lower alcohol is not preferable because the yield of the dye component decreases.

  If sugar removal is incomplete and sweetness remains in the eluent, the evaporation residue is mixed with pure lower alcohol such as pure ethanol, insoluble sugar is filtered off and removed, and the pigment component is removed from the filtrate. By recrystallizing, a highly pure pigment component can be obtained.

  Further, as a method for obtaining the dye component, in addition to the above-described method, the crude brown sugar is directly immersed or digested in pure lower alcohol such as methanol or pure ethanol, and the insoluble sugar content is removed by a method such as filtration. A method of obtaining a pigment component by evaporating the liquid to dryness can also be used. The pigment component of such crude brown sugar can be obtained from the raw crude brown sugar in a yield of about 0.05 to 0.3% by weight, is a light brown hygroscopic powder, has a slightly scorching smell, Is a slightly bitter powder.

The eluate (pigment component) thus obtained is evaporated to dryness, and then purified and separated with a silica gel column to obtain a brown sugar extract.
In addition, as a brown sugar extract, Coct Oligo Pure (trade name, manufactured by Osaka Pharmaceutical Research Laboratories) can be used.

Next, a method for extracting the red bean seed coat extract will be described.
The red bean is called Vigna angularis, and is an annual plant of the leguminous cowpea genus native to East Asia. Usually seeds are edible.
In the present invention, commercially available red beans can be used.

  Immerse the red beans in boiling water. Thereafter, the seed coat is separated by grinding, and the separated seed coat is dried to obtain a red bean seed coat.

The extraction solvent used when obtaining the red bean seed coat extract from the obtained red bean seed coat is not particularly limited, and a hydrophilic organic solvent such as water or alcohol or a mixed solution thereof can be suitably used. Examples of the hydrophilic organic solvent include methanol or ethanol, propyl alcohol, isopropyl alcohol, butanol, isobutanol, etc., but ethanol is preferred.
Examples of the extraction method include a cold immersion method and a digestion method, but are not limited thereto.

  The extract may be used as it is, but the extract may be concentrated to dryness under reduced pressure or the extract may be removed by lyophilization, and the dried product may be used as the red bean seed coat extract.

  The form of brown sugar extract and red bean seed coat extract according to the present invention may be any form such as liquid, solid, powder, paste, gel, etc., and the optimum form for inclusion in the cosmetics to be blended Can be arbitrarily selected.

  Cosmetics containing the brown sugar extract and red bean seed coat extract according to the present invention include lotions, milky lotions, beauty essences, general creams, cleansing creams, facial cleansers, packs, shaving creams, sun creams, sunscreen creams , Suntan lotion, sunscreen lotion, cosmetic soap, foundation, funny, powder, lipstick, lip balm, eyeliner, eye cream, eye shadow, mascara, bath cosmetics, shampoo, rinse, hair dye, hair cosmetics, etc. Is available.

  The blending ratio of the brown sugar extract and the red bean seed coat extract according to the present invention can be arbitrarily changed depending on the cosmetics to be contained, but it is preferable to blend the brown sugar extract: red bean seed coat extract at a weight ratio of 1: 1. This is because the inhibitory action of hyaluronidase is high.

  The blending amount of the brown sugar extract and red bean seed coat extract according to the present invention is preferably 0.001% to 0.5% by weight, and more preferably 0.001% to 0.1% by weight. When the amount is less than 0.001% by weight, the inhibitory action of hyaluronidase is not sufficiently exhibited, and even if the amount exceeds 0.5% by weight, no further effect can be expected, which is not preferable in any case.

  The cosmetic composition according to the present invention may contain a whitening agent, an active oxygen remover, an antioxidant, and an ultraviolet ray inhibitor in addition to the brown sugar extract and the red bean seed coat extract.

  Examples of whitening agents include vitamin Cs and derivatives thereof, hydroquinone and derivatives thereof, cysteine and derivatives thereof, grabrizine, grabrene, liquiritin, and isoquiritin.

  Examples of the active oxygen scavenger include mannitol, β-carotene, astaxanthin, rutin and derivatives thereof.

  Examples of antioxidants include vitamin E and its derivatives, dibutylhydroxytoluene.

  As UV inhibitors, homomenthyl salicylate, 2-methoxyhexycin 2-ethylhexyl, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenonesulfonic acid, sodium 2-hydroxy-4-methoxybenzophenonesulfonate 4-t-butyl-4′-methoxy-dibenzoylmethane, titanium oxide, zinc oxide, cerium oxide.

  EXAMPLES Hereinafter, although this invention is demonstrated in detail based on an Example, this invention is not limited to a following example.

<Method for measuring hyaluronidase inhibitory activity>
A sample solution for examining hyaluronidase inhibitory activity was prepared as follows.

(Sample preparation)
(Brown sugar extract)
5 kg of Okinawan brown sugar is dissolved in 25 L of water, poured into a column with an inner diameter of 8 cm, in which polystyrene resin (Amberlite XAD, 2500 g) is dispersed and packed in 1 L of water, and allowed to flow down at a rate of 20 mL / min. The dye component was adsorbed. Next, water was allowed to flow down and washed with water until there was no sweetness, the sugar content was sufficiently removed, 20% methanol was injected, and flowed down at a rate of 10 mL / min to elute the dye from the adsorbent. The flow-down was continued until no color was observed, and the effluent was mixed and evaporated to dryness under reduced pressure at 60 ° C. or lower to obtain about 16 g of brown residue having no sweetness. This was dissolved in 2 L of pure ethanol, heated, cooled, and the precipitated turbid material was filtered off and concentrated under reduced pressure at 60 ° C. or lower, and the residue was dried at 60 ° C. or lower to give a brown powder having no sweetness. Got. The obtained brown powder was further purified and separated by silica gel chromatography to obtain a brown sugar extract.

(Azuki bean seed extract)
After washing the beans with water, immerse them in boiling water for about 10 minutes. Thereafter, the red bean seed coat was separated and dried by grinding.
100 g of dried red bean seed coat was immersed in 500 g of a 30 wt% aqueous ethanol solution at 40 ° C. for 2 hours, followed by filtration to obtain a first extract. The same extraction operation was performed on the red bean seed coat after the first extraction operation to obtain a second extract. The first and second extracts were combined, ethanol and water were removed by concentration to dryness or lyophilization under reduced pressure, and 12.7 g of the resulting solid was used as an adzuki bean seed coat extract.

(Bagasse extract)
About 100 g of bagasse dried from the residue after pressing the sugarcane, 2 L of 50 wt% ethanol was extracted at 55 ° C. for 2 hours, followed by filtration to obtain a first extract. The bagasse residue after the first extraction operation was subjected to the extraction operation under the same conditions to obtain a second extract. The first and second extracts were combined, ethanol and water were removed by concentration to dryness or lyophilization under reduced pressure, and 5.98 g of the resulting solid was used as a bagasse extract.

(Geo extract)
As the Ziou extract, Nippon Ziyu Pharmaceutical Co., Ltd., Ziou dry extract was used.

(Preparation of sample solution)
2 mg of the sample was weighed and dissolved in 2 ml of an appropriate solvent to prepare 1 mg / mL. Distilled water was used as a solvent for dissolving the sample, but if the sample did not dissolve, it was dissolved in 10% DMSO aqueous solution or 10% methanol aqueous solution and used.
At the time of use, this solution was used by diluting so that the final concentrations of the respective substances were 200 ppm, 600 ppm and 1000 ppm. That is, when the brown sugar extract + red bean seed coat extract is described as 200 ppm, it indicates that the final concentration of the brown sugar extract is 200 ppm and the final concentration of the red bean extract is 200 ppm.

There are the following four types of sample solutions.
Sample liquid (1) Brown sugar extract sample liquid (2) Brown sugar extract + bagasse extract sample liquid (3) Brown sugar extract + Diou extract sample liquid (4) Brown sugar extract + red bean seed coat extract

  The reagent used for measuring the hyaluronidase inhibitory activity was prepared as follows.

(Preparation of reagents)
(0.1 M acetate buffer (pH 4.0))
A 0.2 M sodium acetate solution was mixed with 0.2 M acetic acid at a volume ratio of 82:18, and further diluted 2 times.
(Enzyme hyaluronidase solution)
6 mg of hyaluronidase (Type-4S from Bovine Tests, manufactured by Sigma) is weighed and dissolved in 1.5 ml of 0.1 M acetate buffer (pH 4.0) to 4 mg / ml (final concentration 0.8 mg / ml). It was prepared as follows.
(Enzyme Activator Compound 48/80)
Compound 48/80 (manufactured by Sigma) was weighed 2.5 mg and dissolved in 5 ml of 0.1 M acetate buffer (pH 4.0) to a final concentration of 0.5 mg / ml (final concentration 0.1 mg / ml). Prepared.
(Substrate hyaluronate solution)
10 mg of sodium hyaluronate was weighed and dissolved in 12.5 ml of 0.1 M acetate buffer (pH 4.0) to prepare 0.8 mg / ml (final concentration 0.16 mg / ml).
(P-dimethylaminobenzaldehyde solution)
5 g of p-dimethylaminobenzaldehyde was dissolved in 6 ml of 10N hydrochloric acid and 44 ml of acetic acid to prepare. In use, this solution was diluted 10-fold with glacial acetic acid.
(1M boric acid solution)
61.83 g of boric acid (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in distilled water and dissolved in 1 L.

(Measuring method)
100 μl of the sample solution was added to the test tube, 50 μL of the enzyme hyaluronidase solution was further added, and the mixture was incubated at 37 ° C. for 20 minutes. Thereafter, 100 μL of the enzyme activator Compound 48/80 solution was added, and further incubated at 37 ° C. for 20 minutes, 250 μL of the substrate sodium hyaluronate solution was added, the aluminum cap was applied, and the mixture was reacted at 37 ° C. for 40 minutes.
After stopping the reaction by adding 100 μL of 0.4N sodium hydroxide solution, 100 μL of 1M boric acid solution was added and boiling treatment was performed for 3 minutes. After cooling the reaction solution to room temperature, 3 mL of a p-dimethylaminobenzaldehyde solution diluted 10-fold with glacial acetic acid was added, incubated again at 37 ° C. for 20 minutes, transferred to a microplate, and the absorbance at 585 nm was measured using a microplate reader ( Bio-Rad).
As a control, a sample added with 0.1 M acetate buffer (pH 4.0) instead of the enzyme hyaluronidase solution and a sample solvent added instead of the sample solution were treated in the same manner, and the absorbance was measured.
As a positive control, dipotassium glycyrrhizinate having hyaluronidase inhibitory activity was used at a concentration of 0.5 mg / mL (final concentration 100 ppm).
As a result, the hyaluronidase inhibition rate was calculated according to the following formula.
(formula)
Inhibition rate (%) = (1− (S−SB) / (C−CB)) × 100

S: Absorbance when the sample solution is added (test absorbance)
SB: Absorbance (sample absorbance) when 0.1 M acetate buffer (pH 4.0) was added instead of the enzyme hyaluronidase solution
C: Absorbance when the solvent in which the sample is dissolved is added instead of the sample solution (maximum absorbance)
CB: Absorbance when 0.1M acetic acid buffer (pH 4.0) is added instead of the enzyme hyaluronidase solution and the solvent in which the sample is dissolved is added instead of the sample solution (sample solvent absorbance)

(result)
The results are shown in FIG.
When only brown sugar extract was added, even when 1000 ppm was added, it did not show hyaluronidase inhibitory action, whereas when brown sugar extract 1000 ppm and red bean seed extract 1000 ppm were added, hyaluronidase was inhibited by 70% or more. Confirmed to do.
Next, when the hyaluronidase inhibitory action of only the red bean seed coat extract was examined, the inhibition rate was about 30% even when 1000 ppm was added (see FIG. 2).
From the above results, the mixture of brown sugar extract and red bean seed coat extract according to the present invention has increased hyaluronidase inhibitory action compared to the case where each was added alone, so the combination of brown sugar extract and red bean seed coat extract is related to the hyaluronidase inhibitory action. It was shown that there is a synergistic effect.

  Therefore, the composition containing brown sugar extract and red bean seed coat extract inhibits hyaluronidase and makes it difficult to break down hyaluronic acid, thus moisturizing the skin, improving and preventing wrinkles and sagging and suppressing inflammation. It can be said that there is.

  The formulation example and manufacturing method of the cosmetics which mix | blended the brown sugar extract and the red bean seed coat extract are shown below.

Ａ、Ｂ共に８０℃で加温溶解し、Ａをホモミキサーで攪拌しながらＢを徐々に加えて乳化する。攪拌を続けながら冷却し、４０℃になったところでＣを加え、約３５℃で調製を終了する。 (Formulation Example 1) Cream

Both A and B are heated and dissolved at 80 ° C., and B is gradually added and emulsified while A is stirred with a homomixer. Cool while continuing to stir, and at 40 ° C, add C and finish the preparation at about 35 ° C.

Ａ、Ｂ共に８０℃で加温溶解し、Ａをホモミキサーで攪拌しながらＢを徐々に加えて乳化する。攪拌を続けながら冷却し、４０℃でＣを加え、約３５℃で調製を終了する。 (Prescription Example 2) Emulsion

Both A and B are heated and dissolved at 80 ° C., and B is gradually added and emulsified while A is stirred with a homomixer. Cool while continuing to stir, add C at 40 ° C. and finish the preparation at about 35 ° C.

Ａ、Ｂを室温で溶解する。Ａを攪拌しながらＢを加え調製を終了する。 (Formulation Example 3) Lotion

A and B are dissolved at room temperature. Add B while stirring A to complete the preparation.

Ａを８０℃で攪拌しながら均一に混合する。攪拌しながらＢを加え、３０℃まで冷却し自然脱泡させて作製する。 (Prescription Example 4) Shampoo

A is mixed uniformly at 80 ° C. with stirring. Add B while stirring, cool to 30 ° C., and degas naturally.

水を８０℃で攪拌しながら、上記処方例の表に記載の化合物を上から順番に加えて均一に混合した後３０℃まで冷却して作製する。 (Formulation example 5) Conditioner treatment

While stirring water at 80 ° C., the compounds described in the table of the above formulation examples are added in order from the top and mixed uniformly, and then cooled to 30 ° C. to prepare.

  The present invention provides a hyaluronidase inhibitor and cosmetic composition that contains brown sugar extract and red bean seed coat extract, moisturizes the skin, improves and prevents wrinkles, sagging and inflammation, and is excellent in skin safety. can do.

Claims (3)

  Hyaluronidase inhibitor characterized by containing brown sugar extract and red bean (Vigna angularis) seed coat extract.   A cosmetic composition comprising the hyaluronidase activity inhibitor according to claim 1.   The cosmetic composition according to claim 2, further comprising at least one of a whitening agent, an active oxygen scavenger, an antioxidant, and an ultraviolet light inhibitor.

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