JP2007320950A - Hyaluronidase inhibitor and skin care composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a hyaluronidase inhibitor having excellent hyaluronidase inhibiting effect. <P>SOLUTION: The hyaluronidase inhibitor contains, as an effective component, a mixture prepared by combining a solvent extract from the rootstock of Bergenia ligulata (scientific name: Bergenia ligulata (Wall.) Engl.) and γ-polyglutamic acid having a molecular weight of ≤200,000. The skin care preparation including the mixture has a hyaluronidase-inhibiting action more excellent than that in the single use of γ-polyglutamic acid or the rootstock of Bergenia ligulata. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a hyaluronidase inhibitor and a skin external preparation composition containing the same.

  Hyaluronidase is a hydrolase of hyaluronic acid and is widely distributed in animal tissues. Hyaluronic acid serving as a substrate is a kind of mucopolysaccharide that is abundant in tissues such as skin and joint fluid, and plays a role in the skin such as protecting cells, retaining tissue moisture, and maintaining flexibility. In addition, it plays the role of maintaining the tissue structure / function and maintaining lubricity as a joint fluid.

  The amount of hyaluronic acid in the living body in skin, joints, etc. decreases due to aging, etc., and causes dry skin, rough skin, reduced elasticity, increased wrinkles, or joint pain. In such a state, the hyaluronidase inhibitor suppresses the degradation of hyaluronic acid and contributes to the maintenance of the amount of biological hyaluronic acid.

  On the other hand, hyaluronidase is also known as a pro-inflammatory enzyme. Hyaluronidase is activated during inflammation, destroys the matrix of connective tissue, enhances invasion of inflammatory tissue and vascular permeability, type I allergy It is said that it is likely to mediate in the process of histamine release from mast cells. Therefore, high hyaluronidase inhibitory activity has been observed in indomethacin, aspirin, sodium cromoglycate and the like, which are anti-inflammatory agents and antiallergic agents. Hyaluronidase inhibitors have been studied for the reduction of atopic dermatitis and its associated itching in recent years, the reduction of skin inflammation of sensitive skin, and the prevention and improvement of hay fever.

  Acne that occurs frequently in puberty occurs when acne bacteria proliferate with the increase in sebum secretion. Acne bacteria are known to produce enzymes such as lipase, hyaluronidase, and protease. Inflammation caused by hyaluronidase causes inflammatory acne such as papules and pustules. Therefore, suppressing hyaluronidase activity prevents inflammatory reaction and is considered effective for acne.

As described above, glycyrrhizic acid, sodium cromoglycate, baicalin, indomethacin, aspirin and the like have been confirmed to have high hyaluronidase activity inhibitory activity and are currently used as anti-inflammatory agents. However, these have problems in that there are limitations on the use and use concentration, and side effects are recognized. On the other hand, in order to overcome such problems, a search for a substance having hyaluronidase inhibitory activity from natural products has also been carried out. Examples of substances having hyaluronidase inhibitory activity include rosemary, thyme and Melissa extract (Patent Document 1). ), Saruogase family lichen red snow tea (Patent Document 2), Omega Sasso (Patent Document 3), etc. have been proposed, but the effects and effects are not always sufficient, and effective results are obtained when blended into products. It has not reached.
JP 08-333267 JP2003-212789 JP-A-2005-29556

  The present invention is a hyaluronidase inhibitor and an external preparation for skin intended to suppress inflammation and improve trouble-free skin by applying an inhibitor having a hyaluronidase inhibitory action.

  The present invention relates to Bergenia regulata Bergenia ligulata (Wall.) Engl. The present invention provides a hyaluronidase inhibitor having a synergistic effect by combining a rhizome solvent extract and γ-polyglutamic acid, and a skin external preparation composition containing these.

  The hyaluronidase inhibitor of the present invention has a hyaluronidase inhibitory action superior to the use of γ-polyglutamic acid and Bergenia ligurata alone, and the skin external preparation containing these prevents skin irritation, beautiful skin The state can be maintained.

  Hereinafter, the configuration of the present invention will be described in detail. Bergenia regulater used in the present invention, Bergenia ligulata (Wall.) Engl. Is a plant belonging to the genus Himalaya Yukinoshita, which grows naturally in the northern part of India from the Himalayas. It has been used in Ayurveda, a traditional Indian medicine, to treat cough, fever, diarrhea, and so on. The use site of the present invention is the rhizome. Previously, Bergenia regratata has been reported in patents to prevent aging and eliminate active oxygen. (JP 2004-315492, JP 2004-315491)

  Further, γ-polyglutamic acid used in the present invention is regarded as a main element of the stringing component and the sticky component that are characteristic of natto. Research and utilization of this γ-polyglutamic acid has been conducted for more than 10 years. Water retention is known and used in cosmetics and the like. (JP-A-2004-51533, JP-A-2002-145723, etc.) However, the molecular weight of γ-polyglutamic acid used in the present invention is 200,000 or less, and a hyaluronidase inhibitory action has not yet been found.

  The method for preparing the extract used in the present invention is not particularly limited, and examples thereof include a method of extracting from low temperature to warm using various solvents.

  Specific examples of the extraction solvent include water, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol and other lower monohydric alcohols, glycerin, propylene glycol, dipropylene glycol, and 1,3-butylene. Examples include liquid polyhydric alcohols such as glycol, lower alkyl esters such as ethyl acetate, hydrocarbons such as benzene, hexane, and pentane, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether, tetrahydrofuran, dipropyl ether, and acetonitrile. In addition, one or a mixture of two or more of these can be used.

  The extract used in the present invention may be used as it is, but may be filtered and concentrated as necessary. The extract can also be used after being fractionated and purified by a column chromatography method, a countercurrent distribution method or the like.

  Furthermore, after the above-mentioned product is dried under reduced pressure or freeze-dried, it can be prepared in the form of powder or paste, and can be appropriately formulated and used.

(Production Example 1)
2 kg of 50 vol% ethanol solution is added to 100 g of rhizomes of Bergenia regulata, followed by stirring and extraction at 50 ° C. for 5 hours, cooling and filtration to obtain about 1800 g of extract. The solid content of this extract is adjusted to 1.0% to obtain Extract A.

(Production Example 2)
Using γ-polyglutamic acid adjusted to 1.0% with distilled water as 1, mixtures A and B in which the extract A is mixed at a ratio of 0.01 and 0.1, respectively, are obtained.

(Production Example 3)
3 kg of purified water is added to 200 g of rhizome of Bergenia regulata and extracted with stirring at 80 ° C. for 5 hours. After cooling, it is filtered to obtain about 2600 g of Extract B. The extract B was concentrated to about 200 g under reduced pressure, then treated with a column filled with the synthetic adsorbent Diaion HP-20, and the 10 vol% ethanol solution eluate, 20 vol% ethanol solution eluate and 30 vol after washing with water. Obtain an eluate of% ethanol solution. Each eluate is concentrated under reduced pressure to 50 g, and 50 g of ethanol is added to obtain extracts E, F, and G.

(Production Example 4)
Using γ-polyglutamic acid prepared to 1.0% with distilled water as 1, mixtures C and D in which the extract G is mixed at a ratio of 0.01 and 0.1, respectively, are obtained.

  As a result of measuring the average molecular weight of γ-polyglutamic acid used in Production Examples 2 and 4, the average molecular weight was 60,000. The measuring method analyzed by HPLC using a gel filtration column, and calculated | required the average molecular weight with the light-scattering detector and the differential refractometer.

(Conditions for molecular weight measurement)
GPCMALS measurement GPC-Light scattering measurement conditions GPC equipment: GPC8020 series (manufactured by Tosoh Corporation)
Light scattering detector: DAWNEOS (manufactured by Wyatt Technology)
Concentration detector: differential refractometer (RI detector)
Light scattering detector laser wavelength: 690 nm
Column: α-M (7.8 mm ID × 30 cm) × 2 manufactured by Tosoh Eluent: 50 mM phosphate buffer (pH 7.0) +0.1 M NaCl
Flow rate: 1.0 ml / min
Injection volume: 100 μl
Column temperature: 40 ° C
Detector temperature: 40 ° C
Sample concentration: 0.2%
Pretreatment: Filtration through a 0.5 μm cellulose acetate filter immediately before measurement

  The extract of the present invention can be used as it is, and can also be blended in hair preparations and skin external preparations, but the blending amount is not particularly specified. Although it varies depending on the type, properties, quality, and expected effect of the product to be blended, it is preferably 0.00001 to 2.5% by weight (hereinafter simply referred to as “%”) in terms of dry solid, especially 0.001 to 1.0% is more preferable. When the extract is used, the concentration of the extract is not limited as long as the content of the dry solid content as the solute is within the above range.

  When the plant extract of the present invention is used for a hair preparation or an external preparation for skin, in addition to the above-described components, if necessary, it is usually within the range not impairing the effects of the present invention, such as cosmetics, quasi drugs, pharmaceuticals, etc. Components used for external preparations for skin, such as surfactants, oils, moisturizers, thickeners, antioxidants, UV protection agents, alcohols, powder components, colorants, fragrances, aqueous components, water, various skin nutrients Etc. can be appropriately blended as necessary.

  Further, plants having a physiological activity such as sequestering agents, antiseptic antibacterial agents, cell activators, sebum secretion regulators, anti-inflammatory agents, astringents, whitening agents, active oxygen inhibitors, antiallergic agents, antiaging agents, etc. An extract, a microbial extract, an extract fraction, a purified product, and the like can be appropriately blended.

  The estrogen-like agent of the present invention and the hair preparation and skin external preparation containing the same are not limited to general skin cosmetics, but include pharmaceuticals, quasi drugs, medicinal cosmetics, etc. It is. The dosage form of the hair preparation and skin external preparation of the present invention may be any of a solubilization system, an emulsification system, a powder dispersion system, a powder system, etc., and the use is also a basic cosmetic such as lotion, emulsion, cream, pack, etc. Makeup cosmetics such as foundation, hair nick, hair cream, hair liquid, hair lotion, pomade, hair restorer, hair cosmetics such as shampoo and rinse, toiletries such as soap and body shampoo, bath preparations, etc.

  Next, although an example is given and explained, the present invention is not limited to these examples.

    (Test Example 1) Mixtures A to D obtained from the production examples were used as samples. As a positive control, cromoglycate sodium (manufactured by Fujisawa Pharmaceutical Co., Ltd.) whose inhibitory activity was already known was used in the test. The test method was performed by applying the Morgan-Elson method.

Add 0.1 mL of hyaluronidase (Sigma, Type IV-S, final enzyme activity 400 NFunit / mL) to 0.2 mL of a solution obtained by diluting an appropriate amount of the sample with 0.1 M acetate buffer (adjusted to pH 3.5). After standing at 37 ° C. for 20 minutes, 0.2 mL of a compound 48/80 (Sigma) acetate buffer solution (0.1 mg / mL) is added as an activator, and the mixture is further left at 37 ° C. for 20 minutes. . To this is added 0.5 mL of potassium hyaluronate (manufactured by Wako Pure Chemical Industries) solution (final concentration 0.4 mg / mL), and the mixture is allowed to stand at 37 ° C. for 40 minutes. Next, after 0.2 mL of 0.4N sodium hydroxide solution was added on ice to stop the reaction, 50 mL of water was added to boric acid solution (4.95 g of boric acid, pH 9. with 1N sodium hydroxide solution). (1), then add water to make 100 mL), mix and heat in a boiling water bath for 3 minutes to inactivate the enzyme. Next, it is cooled to room temperature on ice, p-dimethylaminobenzaldehyde reagent (manufactured by Wako Pure Chemical Industries, Ltd., 10 g of 10N hydrochloric acid solution 12.5 mL and acetic acid 87.5 mL are mixed and dissolved, and immediately before use, 10 times with acetic acid. 6 mL) is added and allowed to stand at 37 ° C. for 20 minutes, and then the absorbance is measured at 585 nm. In addition, a sample containing an acetate buffer instead of a sample solution is used as a control, and each sample solution and control is used as a blank containing no enzyme. The inhibitory activity rate is obtained by the following formula, and the sample concentration is adjusted to 50%. The inhibitory activity concentration (IC ₅₀ ) was determined.
Inhibition rate (%) = [1- (absorbance of sample solution−absorbance of sample solution blank) / (absorbance of control solution−absorbance of control solution blank)] × 100

(Test results)
As shown in the test results, a synergistic effect was observed when combined with the inhibitory action alone, and a clear high inhibitory action was confirmed.

  Furthermore, the formulation example of this invention is shown below.

(Example 1) Cream The following components (1) to (10), and separately the following components (11) to (15) are heated and dissolved at 75 ° C. to obtain A solution and B solution, respectively. Liquid B was added to liquid A, emulsified, cooled to 50 ° C. with stirring, and component (16) was added to prepare a cream.
(Ingredient) (wt%)
(1) Jojoba oil 3.0%
(2) Squalane 2.0%
(3) Methyl polysiloxane 0.5%
(4) Stearyl alcohol 0.5%
(5) Cetyl alcohol 0.5%
(6) Tri (capryl / capric acid) glyceryl 12.5%
(7) Glyceryl monostearate 5.0%
(8) 1.5% diglyceryl monostearate
(9) Decaglyceryl monostearate 3.0%
(10) Propyl paraoxybenzoate 0.1%
(11) Xanthan gum 0.1%
(12) Mixture B 2.5%
(13) 1,3-butylene glycol 2.5%
(14) Methyl paraoxybenzoate 0.2%
(15) 66.0% purified water
(16) Fragrance 0.1%

(Example 2) Toner lotion The following components (5) to (8) are mixed and dissolved to give solution A, and separately, the following components (1) to (4) and (9) are mixed and dissolved to give solution B, A liquid and B liquid were mixed uniformly and the lotion was adjusted.
(Ingredient) (wt%)
(1) Glycerol 3.0%
(2) 1,3-butylene glycol 5.0%
(3) Mixture A 10.0%
(4) Polyoxyethylene sorbitan laurate 1.2%
(5) Ethyl alcohol 3.0%
(6) Methyl paraoxybenzoate 0.2%
(7) Fragrance 0.1%
(8) 77.5% purified water

(Example 3) Emulsion The following components (1) to (10), and separately (11) to (14) and (16) are heated and dissolved at 75 ° C. to make A liquid and B liquid, respectively. In addition, the mixture was emulsified, cooled to 50 ° C. with stirring, added with component (15), and an emulsion was prepared.
(Ingredient) (wt%)
(1) Jojoba oil 1.0%
(2) Squalane 2.0%
(3) Behenyl alcohol 1.0%
(4) Tri (capryl / capric acid) glyceryl 2.0%
(5) Tetraglycerin condensed silinoleic acid 0.1%
(6) Propylene glycol monooleate 0.5%
(7) Glycerol monostearate 1.0%
(8) Hexaglyceryl monomyristate 1.0%
(9) Decaglyceryl monomyristate 0.5%
(10) Propyl paraoxybenzoate 0.1%
(11) Mixture C 3.0%
(12) 1,3-butylene glycol 3.0%
(13) Methyl paraoxybenzoate 0.1%
(14) Fragrance 0.1%
(15) Purified water 84.6%

(Example 4) Cleansing Gel The following components (1) to (3), and separately (4) to (6) and (8) were heated and dissolved at 70 ° C. to make A liquid and B liquid, respectively. And stir until uniform. While stirring, the mixture was cooled to 50 ° C. and component (7) was added to prepare a cleansing gel.
(Ingredient) (wt%)
(1) Hexaglyceryl monomyristate 20.0%
(2) Liquid paraffin 59.7%
(3) Paraoxybenzoic acid ester 0.3%
(4) Mixture D 5.0%
(5) Concentrated glycerin 5.0%
(6) Sorbitol 5.0%
(7) Fragrance 0.1%
(8) Purified water 4.9%

(Example 5) Packing agent A phase, B phase, and C phase are each uniformly dissolved, solubilized by adding B phase to A phase, and then uniformly dissolved by adding C phase.
(Ingredient) (wt%)
(Phase A) Dipropylene glycol 5.0%
Polyoxyethylene hydrogenated castor oil 5.0%
(Phase B) Olive oil 5.0%
Tocophenol acetate 0.2%
P-Hydroxybenzoate 0.2%
(Phase C) Sodium bisulfite 0.03%
Polyvinyl alcohol 13.0%
Mixture A 1.0%
Ethanol 7.0%
Purified water 63.77%

(Example 6) Emulsification type foundation A powder part obtained by sufficiently mixing and pulverizing the following components (1) to (6) is A, (7) and (8) are B liquids, and (9) to (12) and (14) Is liquid C. After the liquid C is heated and stirred, A is added and the mixture is homomixed, and the liquid B which is superheated and mixed is added and the mixture is homomixed. Cool to 50 ° C. with stirring, add (13), and further cool to room temperature.
(Ingredient) (wt%)
(1) Titanium dioxide 10.3%
(2) Sericite 5.4%
(3) Kaolin 3.0%
(4) Yellow iron oxide 0.7%
(5) Bengala 0.4%
(6) Black iron oxide 0.2%
(7) Decamethylcyclopentasiloxane 11.5%
(8) Liquid paraffin 8.5%
(9) Sorbitan sesquioleate 3.0%
(10) Mixture B 1.5%
(11) 1,3-butylene glycol 5.0%
(12) Paraoxybenzoic acid ester 0.2%
(13) Fragrance 0.2%
(14) Purified water 50.1%

(Example 7) Solid foundation The following components (1) to (7) are uniformly mixed with a blender, (8) to (14) are added thereto, and the mixture is thoroughly kneaded to prepare.
(Ingredient) (wt%)
(1) Talc 41.9%
(2) Kaolin 18.5%
(3) Sericite 14.0%
(4) Titanium dioxide 3.8%
(5) Yellow iron oxide 2.9%
(6) Bengala 0.5%
(7) Black iron oxide 0.2%
(8) Squalane 8.0%
(9) Isostearic acid 4.0%
(10) Polyoxyethylene sorbitan monooleate 3.0%
(11) Isocetyl octanoate 2.0%
(12) Mixture D 1.0%
(13) Paraoxybenzoic acid ester 0.1%
(14) Fragrance 0.1%

(Example 8) Hair artic (1) to (4) and (7) are added to the following component (5) and dissolved by stirring, and then (6) and (8) are added and further stirred.
(Ingredient) (wt%)
(1) Glycerol 3.0%
(2) L-menthol 0.1%
(3) Assembly extract 2.0%
(4) Mixture C 5.0%
(5) Ethanol 53.0%
(6) Fragrance 0.2%
(8) Purified water 36.7%

(Example 9) Shampoo The following components are heated and mixed uniformly.
(Ingredient) (wt%)
(1) N-coconut oil fatty acid glutamic acid triethanol 25.0%
Amine (2) Lauric acid diethanolamide 5.0%
(3) Potassium myristate 5.0%
(4) Ethylene glycol distearate 2.0%
(5) Polyethylene glycol 400 15.0%
(6) Jojoba oil 1.0%
(7) Mixture D 3.0%
(8) Chloroxylenol 0.1%
(9) Vitamin E 0.1%
(10) Paraoxybenzoic acid ester 0.2%
(11) Fragrance 0.3%
(12) 43.3% purified water

(Example 10) Bath agent (Ingredient) (wt%)
(1) Dry sodium sulfate 40.0%
(2) Sodium bicarbonate 57.5%
(3) Olive oil 0.2%
(4) Mixture C 0.1%
(5) Light anhydrous silicic acid 0.3%
(6) Fragrance 1.7%
(7) Yellow No. 202 (1) 0.2%

(Test example 2) Use effect test The effect test at the time of using the skin external preparation of this invention was done. The usage test was 20 women (21-43 years old) suffering from dryness, wrinkles, and non-skinned skin as a panelist, twice daily in the morning and evening, and 10 people after the face washing for 3 months. It was applied for a while. As a control, 10 people used the mixture C with purified water. The evaluation method was performed according to the following criteria, and the results are shown in Table 2. No one complained of skin abnormalities during use.

"Fine detail"
Effective: Slightly improved foundation paste: Slightly improved foundation paste Invalid or unknown: No change or unknown before use

Claims (2)

  A hyaluronidase inhibitor comprising, as an active ingredient, a composition for external use in which a solvent extract of Bergenia ligrata (scientific name: Bergenia ligurata (Wall.) Engl.) And γ-polyglutamic acid having a molecular weight of 200,000 or less are used in combination.   A topical skin preparation comprising the hyaluronidase inhibitor according to claim 1.