JP2007297559A - AMINO ACID-MODIFIED-(gamma-POLYGLUTAMIC ACID) OR ITS SALT AND THEIR USES - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an amino acid-modified-(γ-polyglutamic acid) having a high moisture-retaining property and new functions and its salt, to provide a method for producing the compound, and to provide a use thereof. <P>SOLUTION: This amino acid-modified-(γ-polyglutamic acid) or its salt is characterized by amide-bonding a part of all of the side chain α-carboxyl groups ofγ-polyglutamic acid to the amino group of a hydrophilic amino acid. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to amino acid modification (γ-polyglutamic acid) having a new function by modifying γ-polyglutamic acid with a hydrophilic amino acid.

  γ-polyglutamic acid is derived from a polymer of glutamic acid ester-N carboxylic acid anhydride, or produced by fermentation of microorganisms belonging to the genus Bacillus, and is a water-soluble product in which a plurality of glutamic acids are linked by γ-glutamyl bonds It is a biodegradable polymer. Since the salt of γ-polyglutamic acid is excellent in moisturizing property, polyglutamate is particularly useful as a moisturizing agent for various products (shoe creams, paints, paper products, foods, cosmetics, etc.) that require moisturizing properties. Since it is a kind of polypeptide that is a component of NMF (natural moisturizing factor) of skin, its use as a moisturizing agent for cosmetics has been proposed (see Patent Document 1).

  On the other hand, since γ-polyglutamic acid has a carboxyl group derived from the α-carboxyl group of glutamic acid in the side chain, by introducing various functional groups into the side chain carboxyl group, γ-polyglutamic acid is further new to γ-polyglutamic acid. Many modifications with functions (γ-polyglutamic acid) have been proposed. For example, a compound in which taurine having a sulfoalkylamino group is bonded to a side chain carboxyl group of γ-polyglutamic acid to give a new function (Patent Document 2), a polyoxyethylene chain or an alkyl group is bonded to form micelles (Patent Document 3) to which a function is imparted, a temperature responsiveness is imparted by binding amino alcohol (Non-patent Document 1), and a new function is imparted by binding an alkyl group (Non-patent Document 2) And an L-phenylalanine derivative bonded to increase the hydrophobicity and applied to nanoparticles (Non-patent Document 3).

By the way, the moisturizing action is one of the important functions for skin application agents such as external preparations for skin and skin cosmetics and hair care products. Conventionally used moisturizers are mainly polyhydric alcohols such as glycerin, propylene glycol and sorbitol. Recently, pyrrolidone carboxylic acid, which is the main component of natural moisturizing factor NMF (natural moisturizing factor). Salts and lactates, sodium hyaluronate and the like are also used. However, polyhydric alcohol humectants are relatively good in moisturizing properties, but tend to cause stickiness and have a difficulty in use. Pyrrolidone carboxylates and lactates are prone to emulsification inhibition and are not applicable to emulsion forms such as emulsions and creams. In addition to high raw materials, sodium hyaluronate is produced by various methods such as chicken crown, microbial production, and gene recombination, and many safety points are required.
JP 59-209635 A JP 2002-80593 A JP 2003-342367 A Tachibana et al., Polymer Preprints Japan 53 (1), IIIPc125 (2004) M. Morillo et al., Polymer 44, 7557 (2003) M. Matsuzaki et al., Chemistry Letters 33 (4) 398 (2004)

  An object of the present invention is to provide a novel γ-polyglutamic acid derivative [modified- (γ-polyglutamic acid) or a salt thereof] in which a new function is imparted to γ-polyglutamic acid. In particular, an object is to provide a modified (γ-polyglutamic acid) salt that is significantly better in moisture retention than γ-polyglutamate. Another object of the present invention is to provide a method for producing the modification (γ-polyglutamic acid) or a salt thereof, and use of the modification (γ-polyglutamic acid) or a salt thereof.

  The inventors of the present invention have been diligently studied to solve the above-mentioned problems. An amino acid prepared by amide-bonding an amino group of an amino acid having a hydrophilic group to the α-carboxyl group of the side chain of γ-polyglutamic acid It has been found that the salt of modified- (γ-polyglutamic acid) has better moisturizing properties than γ-polyglutamic acid and has the effect of significantly suppressing the adhesion of Staphylococcus aureus to the skin. It was convinced that it was useful not only as a moisturizer but also as a cosmetic preparation and a skin external preparation for skin diseases caused by infection with Staphylococcus aureus such as atopic dermatitis. Furthermore, the present inventors have found that the salt of γ-polyglutamic acid and the above-mentioned amino acid modification- (γ-polyglutamic acid) has an action of suppressing bacterial adhesion to contact lenses, and these compounds are The present inventors have found that it is useful as a compounding component for various contact lens compositions such as contact lens cleaning agents, preservatives, contact lens mounting solutions, and eye drops for mounting contact lenses. The present invention has been developed based on such knowledge.

That is, the present invention includes the following modes.
(1) Amino acid modification— (γ-polyglutamic acid) or a salt thereof (1-1) A part or all of the side chain α-carboxyl group of γ-polyglutamic acid and an amino group of a hydrophilic amino acid are amide-bonded. Amino acid modification- (γ-polyglutamic acid) or a salt thereof.
(1-2) General formula (I)

[In the formula, R is a hydrophilic group represented by the following formula (II),

(In the formula (II), R ₁ represents a lower alkylene group, R ₂ represents a lower alkylene group which may be substituted with a hydroxyl group, and R ₃ represents a hydrogen atom, a hydroxyl group or a carboxyl group. X and y represent , The same or different, indicating 0 or 1.)
m represents an integer of 0 or 1 or more, l represents an integer of 6 to 15000 together with m, and each repeating unit is present randomly or partially forms a block. ]
(1-1) amino acid modification represented by (γ-polyglutamic acid) or a salt thereof.
(1-3) The description of (1-2), wherein the hydrophilic group (II) is a group obtained by removing an amino group from at least one hydrophilic amino acid selected from the group consisting of glycine, serine, aspartic acid and glutamic acid Amino acid modification of-(γ-polyglutamic acid) or a salt thereof.
(1-4) Amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-3) having a molecular weight of 1,000 to 5,000,000.

(2) Amino acid modification— (γ-polyglutamic acid) or a method for producing a salt thereof (2-1) γ-polyglutamic acid and an alkyl ester of a hydrophilic amino acid are converted to 1-ethyl-3- (3-dimethylaminopropyl)- The method for producing amino acid modification- (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-4), comprising a step of reacting in the presence of carbodiimide hydrochloride.
(2-2) It is characterized by reacting γ-polyglutamic acid with an alkyl ester of a hydrophilic amino acid in the presence of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride and 4-dimethylaminopyridine. A method for producing an amino acid modification (γ-polyglutamic acid) or a salt thereof according to (2-1).

(3) Moisturizer (3-1) A moisturizer comprising the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-4) as an active ingredient.

(4) Staphylococcus aureus or Pseudomonas aeruginosa adhesion inhibitor (4-1) γ-polyglutamic acid, amino acid modification according to any one of (1-1) to (1-4)-(γ-polyglutamic acid) And an agent for suppressing the adhesion of Staphylococcus aureus or Pseudomonas aeruginosa, comprising as an active ingredient at least one compound selected from the group consisting of these salts.

(5) External preparation for skin (5-1) An external preparation for skin containing the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-4).

(6) Cosmetics (6-1) Cosmetics containing the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-4).

(7) Hair care agent (7-1) A hair care agent containing the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of (1-1) to (1-4).

(8) Contact lens composition (7-1) γ-polyglutamic acid, amino acid modification according to any of (1-1) to (1-4)-(γ-polyglutamic acid), and salts thereof A composition for contact lenses, comprising at least one compound selected from the group consisting of:

(1) Amino acid modification- (γ-polyglutamic acid) and salts thereof The amino acid modification- (γ-polyglutamic acid) of the present invention [hereinafter also referred to as “amino acid modification-γPGA”] The side chain α-carboxyl group of the linked γ-polyglutamic acid has a structure in which all or part of it is bonded with an amino group of a hydrophilic amino acid.

  The hydrophilic amino acid targeted by the present invention has both an amino group and a carboxyl group, and is hydrophilic as a whole. As long as it has such properties, it is not limited to α-amino acids constituting proteins, and for example, a part or all of the alkyl group may be substituted with a hydroxyl group. The hydrophilic α-amino acids include glycine, serine, threonine, cysteine, asparagine, aspartic acid, glutamine, glutamic acid, lysine, arginine, histidine, 2-aminoadipic acid, 2-aminozberic acid, and 2-aminopimelic acid. And so on. Preferable examples include glycine, serine, aspartic acid, glutamic acid, and 2-aminosuberic acid. The amino acids targeted by the present invention may be β-amino acids and γ-amino acids having amino groups bonded to the 3rd and 4th positions based on the carbon (α-C) to which the carboxyl group is bonded. Good. Specific examples of β-amino acids include 3-aminoadipic acid, 3-aminosuberic acid, and 3-aminopimelic acid. Specific examples of γ-amino acids include 4-aminosuberic acid and 4-aminopimelic acid. can do. These α-, β-, or γ-amino acids may be those in which part or all of the alkyl groups are substituted with hydroxyl groups. Specifically, β-hydroxyaspartic acid, γ- Examples include hydroxyaspartic acid.

  The amino acid targeted by the present invention may be either D-form or L-form. The L form is preferred.

The amino acid modification-γPGA of the present invention modified with these amino acids can be specifically represented by the following general formula (I).
Formula (I)

Here, R means a hydrophilic group represented by the following formula (II).

In the formula (II), R ₁ represents a lower alkylene group, R ₂ represents a lower alkylene group which may be substituted with a hydroxyl group, and R ₃ represents a hydrogen atom, a hydroxyl group or a carboxyl group. Note that x and y are the same or different (independent of each other) and mean an integer of 0 or 1. When x is 0, the carbon at the 2-position (α-C) (indicated by * in the above formula) is directly bonded to the carboxyl group, and when y is 0, the carbon at the 2-position (α-C) Will bind directly to R ₃ . Hereinafter, for convenience, when x is 0, R ₁ is also directly bonded, and when y is 0, R ₂ is directly bonded.

  Here, examples of the lower alkylene group include a linear or branched alkylene group having 1 to 6 carbon atoms, and a linear alkylene group is preferred. Specific examples include a methylene group, an ethylene group, a trimethylene group, a tetramethylene group, a pentamethylene group, and a hexamethylene group. Preferred are a methylene group and an ethylene group.

The lower alkylene group represented by R ₂ may be substituted with a hydroxyl group. The site of the alkylene chain substituted with a hydroxyl group is arbitrary and is not particularly limited. Specifically, depending on the length of the lower alkylene group, the second position, the third position, the fourth position, the fifth position on the basis of the carbon (α-C) (second position) to which the carboxyl group is bonded via R _1. Examples are those in which a hydroxyl group is bonded to any one of the carbon atoms at the 6th, 7th, and 8th positions. The number of hydroxyl groups to be substituted can also be appropriately selected within the range of 1-6. Preferably, it is a lower alkylene group having 1 to 6 carbon atoms which is not substituted with a hydroxyl group or substituted with one hydroxyl group.

Various combinations of R ₁ , R ₂ and R ₃ are not particularly limited, but the hydrophilic group (II) represented by the above formula (II) binds an amino group (—NH ₂ ) and is hydrophilic as described above. Combinations that are amino acids are preferred.

For example, a combination in which x and y are both 0 (when R ₁ and R ₂ are both directly bonded) and R ₃ is a hydrogen atom (group other than -NH ₂ of glycine); x is 0 (R ₁ Is a direct bond), R ₂ is a methylene group, and R ₃ is a hydroxyl group (group except serine —NH ₂ ): x is 0 (R ₁ is a direct bond), R ₂ is a methylene group, R ₃ Is a carboxyl group (group of aspartic acid excluding -NH ₂ ): x is 0 (R ₁ is directly bonded), R ₂ is a hydroxymethylene group, and R ₃ is a carboxyl group (β- Group of hydroxyaspartic acid excluding —NH ₂ ): x is 0 (R ₁ is directly bonded), R ₂ is an ethylene group, and R ₃ is a carboxyl group (a group excluding —NH ₂ of glutamic acid): x is 0 (bond _{R 1} directly), _{R 2} is hydroxyethyl Les Group, the combination _{R 3} is a carboxyl group (beta-or γ- hydroxy groups excluding the -NH ₂ of aspartic acid): x is 0 (bond _{R 1} directly), _{R 2} is a trimethylene group, _{R 3} is carboxyl A combination of groups (excluding -NH ₂ of 2-aminoadipic acid): a combination of R ₁ being a methylene group, R ₂ is an ethylene group, and R ₃ is a carboxyl group (-NH _{2 of} 3-aminoadipic acid Group in which x is 0 (R ₁ is a direct bond), R ₂ is a pentamethylene group, and R ₃ is a carboxyl group (a group in which 2-NH2 of ₂ -aminosuberic acid is excluded): R ₁ is A combination of a methylene group, R ₂ is a tetramethylene group, and R ₃ is a carboxyl group (a group other than -NH _{2 of} 3-aminosuberic acid): R ₁ is an ethylene group, R ₂ is a trimethylene group, and R ₃ is a carboxyl group In Combinations (group excluding the -NH ₂ of 4 Aminozuberin acid): x is 0 (bond _{R 1} directly), _{R 2} is a tetramethylene group, _{R 3} is a combination (2-aminopimelic acid, a carboxyl group -NH ₂ ): R ₁ is a methylene group, R ₂ is a trimethylene group, and R ₃ is a carboxyl group (a group other than -NH _{2 of} 3-aminopimelic acid): R ₁ is an ethylene group, R ₂ Are ethylene groups and R ₃ is a carboxyl group (groups of 4-aminopimelic acid excluding —NH ₂ ).

Preferably, x and y are both 0 (when R ₁ and R ₂ are both directly bonded) and R ₃ is a hydrogen atom (group other than -NH ₂ of glycine); x is 0 (R ₁ is a direct bond), R ₂ is a methylene group, and R ₃ is a hydroxyl group (group other than —NH ₂ of serine): x is 0 (R ₁ is a direct bond), R ₂ is a methylene group, R _A combination in which ₃ is a carboxyl group (a group excluding -NH ₂ of aspartic acid): a combination in which x is 0 (R ₁ is directly bonded), R ₂ is an ethylene group, and R ₃ is a carboxyl group (-NH of glutamic acid ₂ ), R ₂ is a hydroxymethylene group, R ₃ is a carboxyl group (group excluding -NH ₂ of β-hydroxyaspartic acid), x is 0 (R ₁ is directly bonded), R ₂ is a pentamethylene group, _{R 3} is a carboxyl group Certain combinations (group excluding the -NH ₂ of 2 Aminozuberin acid): _{R 1} is a methylene group, _{R 2} is a tetramethylene group, _{R 3} is excluding -NH ₂ combinations (3 Aminozuberin acid is a carboxyl group Group): a combination in which R ₁ is an ethylene group, R ₂ is a trimethylene group, and R ₃ is a carboxyl group (a group other than —NH _{2 of} 4-aminosuberic acid).

More preferably, a combination in which both x and y are 0 (when R ₁ and R ₂ are both directly bonded) and R ₃ is a hydrogen atom (group other than —NH ₂ of glycine); R ₁ is a direct bond), R ₂ is a methylene group, and R ₃ is a hydroxyl group (group other than —NH ₂ of serine): x is 0 (R ₁ is a direct bond), R ₂ is a methylene group, A combination in which R ₃ is a carboxyl group (group of aspartic acid excluding —NH ₂ ): a combination in which x is 0 (R ₁ is directly bonded), R ₂ is an ethylene group, and R ₃ is a carboxyl group (- Group excluding NH ₂ ).

  In general formula (I), the sum of m, which means the number of repetitions of “γ-glutamic acid residue”, and l, which means the number of repetitions of “amino acid bond-γ-glutamic acid residue”, is 6 to 15000, preferably Is an integer from 15 to 7000. m is 0 or an integer of 1 or more (however, it does not exceed 15000), and l is an integer obtained by subtracting the integer of m from the sum of m and l. The molecular weight of the amino acid modification-γPGA is not particularly limited, but is usually 1,000 to 5,000,000, preferably 2,000 to 3,000,000, more preferably 3,000 to 2,000,000. The number of repeating units (m , L) can be set as appropriate. Each repeating unit may partially form a block or may exist randomly.

  As the salt of amino acid modification-γPGA of the present invention, basic salts such as alkali metal salts such as sodium and potassium, alkaline earth metal salts such as calcium and magnesium, ammonium salts, ethanolamine salts, diethanolamine salts, triethanolamine salts, etc. Mention may be made of amine salts. Alkali metal salts such as sodium and potassium are preferred.

  The amino acid modification-γPGA of the present invention is an amino acid ester obtained by protecting all carboxyl groups of an amino acid to be modified by esterification with γ-polyglutamic acid (also simply referred to as “γ-PGA”). It can be prepared by reacting in the presence of 3 dimethylaminopropyl) -carbodiimide.

  The γ-PGA used as the starting material is not limited to the production method such as synthesis method or fermentation method, but the molecular weight is in the range of 1,000 to 5,000,000, preferably 2,000 to 3,000,000, more preferably 3,000 to 2,000,000. It is desirable that it is γ-PGA. Γ-PGA having such a molecular weight can be produced from a microorganism belonging to the genus Bacillus by the method described in Reference Examples described later with reference to the descriptions in Japanese Patent Publication No. 43-24472 and Japanese Patent Application Laid-Open No. 1-174397. In addition, commercially available γ-PGA can be used (for example, Nippon Polygly Co., Ltd., Ichimaru Falcos Co., Ltd., Meiji Food Materials Co., Ltd., etc.).

  Examples of the amino acid ester include a lower alkyl ester having 1 to 6 carbon atoms of an amino acid, preferably a lower alkyl having 1 to 3 carbon atoms, and a methyl ester or an ethyl ester of an amino acid is preferably exemplified. The amino acid ester is preferably used as an acid salt with an organic acid or an inorganic acid, specifically as a hydrochloride. Also, 1-ethyl-3- (3 dimethylaminopropyl) -carbodiimide is preferably used as an acid salt with an organic acid or an inorganic acid, specifically as a hydrochloride salt.

  Here, the ratio of γPGA and amino acid ester used for the reaction is 1 to 1.5 mol, preferably 1.1 to 1.2 mol, of amino acid ester with respect to 1 mol of glutamic acid which is a repeating unit of γ-PGA. A percentage can be mentioned. In this case, 1-ethyl-3- (3 dimethylaminopropyl) -carbodiimide or its acid salt is added to the reaction system in an amount of 1 to 1.5 mol mol, preferably 1.2 to 1.3 mol mol. Is desirable.

  In order to increase the yield of the amino acid modification-γPGA of the present invention, the above reaction is added to 1-ethyl-3- (3 dimethylaminopropyl) -carbodiimide or its acid salt and further in the presence of 4-dimethylaminopyridine. Preferably it is done. 4-dimethylaminopyridine is added to the reaction system at a ratio of 0.05 to 0.15 mol, preferably 0.07 to 0.12 mol, per mol of glutamic acid which is a repeating unit of γPGA. It is desirable to keep it.

  After blending each of these components, the amino acid modification-γPGA ester salt is allowed to react with stirring at a low temperature of 0 to 4 ° C. for about the first 2 hours, and then reacted with stirring for about half a day or one day at room temperature. Produces. By hydrolyzing the ester salt thus formed with an aqueous solution of an alkali metal salt such as sodium hydroxide or potassium hydroxide, an alkali metal salt of amino acid modification-γPGA can be obtained. In addition, the amino acid modification-γPGA ammonium salt or basic amine salt (for example, ethanolamine salt, diethanolamine salt, triethanolamine salt, etc.) is converted into amino acid modification-γPGA (carboxylic acid type) prepared by the method described later. It can be prepared by reacting ammonium or a basic amine. In addition, since this reaction is caused by the coexistence of both, it is not particularly limited. For example, ammonia is passed through an aqueous solution containing amino acid modification-γPGA (carboxylic acid type), or a base such as aqueous ammonia, ethanolamine, diethanolamine, or triethanolamine. A method of adding and concentrating a functional amine, and elution by passing an aqueous solution containing amino acid modification -γPGA (carboxylic acid type) through a cation exchange resin bound with a basic amine such as ethanolamine, diethanolamine or triethanolamine. Examples include a method for recovering the liquid.

  In this case, in order to prepare a salt of amino acid-modified PGA having a weak acid pH suitable for the skin, acid treatment may be performed with hydrochloric acid or the like so that the pH is about 5 after hydrolysis.

  Furthermore, amino acid modification-γPGA (carboxylic acid type) can be prepared by treating the salt of amino acid modification-γPGA obtained above with an acid treatment, preferably with a strong acid having a pH of 2 or less.

  The amino acid modification-γPGA and salts thereof thus obtained can be obtained as a powder form by subjecting to purification treatment such as dialysis as needed and freeze-drying.

(2) Application of moisturizer and external preparation for skin, cosmetics or hair care of the moisturizer The amino acid modification-γPGA and the salt thereof of the present invention described above are excellent as shown in Experimental Example 1 described later. Has a moisturizing action. According to Experimental Example 1, among the amino acid-modified PGA, the moisturizing action of a salt of γ-PGA modified with glutamic acid or serine is significantly superior to a conventionally known moisturizing agent such as hyaluronic acid. The salt of γPGA and aspartic acid modified γPGA and its salt have a moisturizing effect superior to that of acetylhyaluronic acid and γ-polyglutamine having higher moisturizing effect than hyaluronic acid.

Therefore, the amino acid modification-γPGA and salts thereof of the present invention are useful as a humectant. The moisturizing agent may be composed of 100% by weight of the amino acid modification-γPGA of the present invention or a salt thereof, and can be used by being blended with an external preparation for skin, a cosmetic or a hair care agent. Here, the external preparation for skin refers to a composition containing various active ingredients and / or additives, and is mainly used outside the body for pharmaceutical purposes. The cosmetics are compositions containing various active ingredients and / or additives, and are mainly used outside the body for skin care and cosmetic purposes. Furthermore, the hair care agent refers to a composition containing various active ingredients and / or additives, which is mainly used for hair or scalp for the purpose of caring for the hair or scalp or for making hair. These external preparations for skin, cosmetics and hair care agents are classified mainly by active ingredients incorporated therein. Therefore, although it is impossible to clearly distinguish from the product forms, for example as a skin external preparation, solutions, ointments, creams, lotions, emulsions, patches and Pas-up agents, and the like; facial cleanser as cosmetics ( Cream, foam, lotion, milk), body shampoo, cleansing (cream, foam, lotion, milk), marsage cream, cold cream, serum, lotion, milk, moisturizing cream, pack, foundation, lipstick, lip balm, Lip gel, makeup, after shaving cream, sunscreen cream, nail preparation (manicure, pedicure, etc.), hand cream, body lotion (or cream or foam), bath preparation, floccoli sheet, antiperspirant, etc .; for hair care As an agent Chick, hair liquids, mention may be made of hair-setting lotion, hair spray, hair color, hair bleach, permanent agents, hair shampoo, hair rinse, hair treatment, hair cream, hair tonic, and such as hair tonic.

  These external preparations for skin, cosmetics or hair care are prepared in any form (for example, cream, gel, ointment, liquid, emulsion, foam, spray, powder, etc.) depending on the application as described above. be able to. Furthermore, in the case of a bath agent, in addition to the above-mentioned form, it can be provided in the form of a tablet, powder or granule.

  When the moisturizing agent of the present invention is blended with the various products, the moisturizing action can be imparted to the various products according to the amount of the moisturizing agent. The blending ratio of the humectant is not limited, but is preferably 0.0001 to 15% by weight, preferably 0.001 to 10% by weight, preferably in terms of the ratio of the amino acid modification-γPGA or a salt thereof of the present invention. More preferably, the content is 0.01 to 5% by weight.

  In addition to the amino acid modification-γPGA of the present invention or a salt thereof, the above various skin external preparations, cosmetics, or hair care agents may contain other moisturizers as long as the effects of the present invention are not hindered. You can also. Such humectants include polyhydric alcohols such as polyethylene glycol, propylene glycol, glycerin, sorbitol, mannitol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol; sodium hyaluronate, acetyl hyaluronic acid, γ-poly Biopolymers such as glutamic acid, heparin-like substance, sodium chondroitin sulfate, collagen, elastin, keratin; amino acids such as glycine, aspartic acid, arginine; natural moisturizing factors such as sodium lactate, urea, sodium pyrrolidone carboxylate; ceramide, cholesterol, Examples thereof include lipids such as phospholipids; plant extracts such as lavender extract, eucalyptus extract, and peppermint extract.

  In addition to the above moisturizers, for example, anti-inflammatory agents, vitamin agents, antibacterial agents, topical agents for skin external preparations, cosmetics or hair care agents, depending on the purpose and use of pharmaceuticals, cosmetics or quasi drugs. Other medicinal ingredients such as anesthetics and whitening agents can be blended.

  Although not limited, anti-inflammatory agents include glycyrrhizic acid and glycyrrhizic acid derivatives such as dipotassium glycyrrhizinate and monoammonium glycyrrhizinate, glycyrrhetinic acid or its derivatives, allantoin or its derivatives, ε-aminocaproic acid, licorice extract, menthol , Camphor and the like.

  Further, as vitamin agents, retinol derivatives such as retinol, acetic acid retinol, retinol palmitate (vitamins A), retinal, retinoic acid, methyl retinoic acid, ethyl retinoic acid, retinol retinoic acid, vitamin A oil, vitamin A fatty acid ester, etc. Vitamins A, α-carotene, β-carotene, γ-carotene, δ-carotene, lycopene, zeaxanthin, cryptoxanthin, echinone and other provitamins A, dl-α-tocopherol, dl-α-tocopherol acetate, succinate Vitamin E such as acid dl-α-tocopherol and dl-α-tocopherol calcium succinate, riboflavin, flavin mononucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, Vitamin B2 such as riboflavin sodium 5'-phosphate and riboflavin tetranicotinate, dl-α-tocopherol nicotinate, benzyl nicotinate, methyl nicotinate, β-butoxyethyl nicotinate, 1- (4- Nicotinic acids such as methylphenyl) ethyl, ascorbic acid, sodium ascorbate, dehydroascorbic acid, sodium ascorbate phosphate, magnesium ascorbate phosphate, magnesium ascorbate, L-ascorbyl dipalmitate, etc. or vitamin C , Vitamin D such as methyl hesperidin, ergocalciferol, cholecalciferol, vitamin K such as phylloquinone, farnoquinone, γ-oryzanol, dibenzoylthiamine, diben Irtiamine hydrochloride, thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate ester, thiamine monophosphorus Vitamin B1 such as acid ester, thiamine diphosphate ester, thiamin diphosphate hydrochloride, thiamine triphosphate ester, thiamine triphosphate monophosphate, pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, 5'-pyridoxal phosphate, hydrochloric acid Vitamin B6 such as pyridoxamine, vitamin B12 such as cyanocobalamin, hydroxocobalamin, deoxyadenosylcobalamin, folic acid such as folic acid and pteroylglutamic acid, nicotinic acid, nicotine Nicotinic acids such as amide, pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), pantothenic acid such as D-panthecin, D-panthetin, coenzyme A, pantothenyl ethyl ether, and biotin such as biotin and biotisin In addition, vitamin-like agents such as carnitine, ferulic acid, α-lipoic acid and orotic acid can be exemplified.

  Examples of antibacterial agents include isopropylmethylphenol, chlorhexidine hydrochloride, benzalkonium chloride, and cetylpyridinium chloride. Examples of the local anesthetic include eucalyptus oil, eugenol, menthol, camphor, and peppermint oil.

  Furthermore, in addition to the amino acid modification-γPGA or a salt thereof of the present invention, it is usually used depending on the type and form of the skin external preparation, cosmetic or hair care agent, as long as the effects of the present invention are not hindered. Bases or additives can be blended. Examples of such bases and additives include pigments, fragrances, preservatives, pH adjusters, metal sequestering agents, higher alcohols, penetration enhancers, and antioxidants. The pH of these preparations is not limited as long as it does not cause skin irritation, but is in the range of pH 4 to 7, preferably pH 5 to 6, which is a weakly acidic region.

(3) Application of the Staphylococcus aureus adhesion inhibitor and the inhibitor to an external preparation for skin or cosmetics The amino acid modification-γPGA salt and γ-PGA salt of the present invention described above are shown in Experimental Example 2 described later. Thus, it has the effect | action which inhibits the skin adhesion of Staphylococcus aureus.

  Therefore, the amino acid-modified salt of γPGA and the salt of γ-PGA of the present invention can be used as a skin adhesion inhibitor for Staphylococcus aureus. The Staphylococcus aureus skin adhesion inhibitor is impaired or aggravated by the growth of Staphylococcus aureus attached to the skin surface via plasma components exuded from scratches, such as atopic dermatitis It is useful for preventing skin diseases. It is also useful for injuries such as scratches, scratches and cuts where plasma components are exuded to the outside of the body and there is a concern that the above-mentioned damage due to Staphylococcus aureus is aggravated.

  The S. aureus skin adhesion inhibitor may comprise the amino acid modification-γPGA salt or γ-PGA salt 100% by weight of the present invention. Such an inhibitor is practically incorporated into an external preparation for skin, cosmetics or skin care agent to inhibit and prevent the adhesion of Staphylococcus aureus to the skin, or to improve or prevent atopic dermatitis, or It can be put to practical use as an external preparation for skin or cosmetic for preventing suppuration of wounds. Various forms of the external preparation for skin or cosmetics are as described above.

  When formulating as a skin external preparation or cosmetic, the content of the amino acid-modified-γPGA or the salt of γ-PGA in the formulation is not limited as long as the effect of the present invention is exhibited, but usually 0 based on the total amount of the formulation. It is 0.0001 to 15% by weight, preferably 0.001 to 10% by weight, and more preferably 0.01 to 5% by weight.

  The application method may be based on the application method of general skin external preparations, cosmetics, skin care agents, etc., and use by applying an appropriate amount to the outer skin such as skin once to several times a day. Can do.

  By adding a medicinal component other than amino acid modification-γPGA or a salt of γ-PGA to the skin external preparation or cosmetic, the effect of the present invention can be enhanced or further effects can be imparted. Such a medicinal component is not particularly limited as long as it has a medicinal effect on the skin and mucous membranes, and is a component that provides all desirable effects such as healing and improvement of inflammation and damage occurring on the skin and mucous membranes, and skin protection. Say. Specific examples of such medicinal components include anti-inflammatory agents, vitamin agents, moisturizers, antibacterial agents, and local anesthetics. Specific examples thereof are the same as those described above.

  In addition, the external preparation for skin or cosmetics may contain pigments, fragrances, preservatives, surfactants, plant extracts, and pH adjusters according to the types and forms of these products as long as the effects of the present invention are not impaired. Various bases and additives such as sequestering agents, oil components, gelling agents, polyhydric alcohols, higher alcohols, hydrocarbons, penetration enhancers, pigments and antioxidants can also be blended. In addition, the pH of the preparation when the Staphylococcus aureus skin adhesion inhibitor of the present invention is formulated into an external preparation for skin or cosmetics may be any pH that does not cause skin irritation, but is usually pH 4 to 7, preferably It is in the range of slightly acidic to neutral pH 5-7.

(4) Application of the Pseudomonas aeruginosa adhesion inhibitor and the inhibitor to the composition for contact lenses The above-described amino acid modification-γPGA, γ-PGA, or salt thereof of the present invention is shown in Experimental Example 3 described later. Thus, it has the effect | action which inhibits adhesion to the contact lens of Pseudomonas aeruginosa. Among them, γ-PGA and salts of γ-PGA modified with glycine or aspartic acid have remarkably excellent Pseudomonas aeruginosa adhesion inhibitory action.

  For this reason, the amino acid modification-γPGA or γ-PGA salt of the present invention can be used as a contact lens adhesion inhibitor of Pseudomonas aeruginosa. Such a contact lens adhesion inhibitor of Pseudomonas aeruginosa can prevent Pseudomonas aeruginosa contamination of the contact lens and prevent eye diseases (for example, corneal tumors) caused by Pseudomonas aeruginosa infection associated with the use of the contact lens.

  The contact lens adhesion inhibitor of Pseudomonas aeruginosa may comprise 100% by weight of the amino acid-modified γPGA or γPGA salt of the present invention. Such an inhibitor can be practically used as a highly antibacterial contact lens composition having an inhibitory effect on adhesion of Pseudomonas aeruginosa to a contact lens by blending in the contact lens composition. .

  Here, examples of the contact lens composition include a disinfectant for a contact lens, a preservative, a cleaning agent, a cleaning preservative, a protein removing agent, a contact lens wearing agent, and an eye drop or eye wash when wearing a contact lens. it can. The contact lens can be applied to any contact lens such as a hard contact lens, an oxygen permeable contact lens, and a soft contact lens regardless of the type of contact lens.

  The blending ratio of the contact lens adhesion inhibitor of Pseudomonas aeruginosa with respect to these contact lens compositions is not particularly limited as long as it exerts the action of inhibiting the adhesion of Pseudomonas aeruginosa, but for example, the composition for contact lenses is In the case of a contact lens disinfectant, preservative, cleaning agent, cleaning preservative, or protein removing agent, it is usually 0.0001 to 2% by weight in terms of the ratio of amino acid modification-γPGA or γ-PGA salt, Preferably in the range of 0.001 to 1% by weight, more preferably in the range of 0.01 to 0.1% by weight, when the contact lens composition is a contact lens wearing agent, or an eye drop or eye wash when wearing a contact lens Is usually 0.0001 to 1% by weight, preferably 0.001 to 0.5% by weight, more preferably in terms of the ratio of amino acid modification-γPGA or γ-PGA salt It can be exemplified range of .01～0.1 wt%.

  In the contact lens composition of the present invention, other components can be blended in accordance with the type of contact lens composition and its use within a range not departing from the object of the present invention.

  For example, if the contact lens composition is a contact lens disinfectant, preservative, cleaning agent, cleaning preservative, or protein remover, in addition to the base (eg, distilled water, sterile purified water, physiological saline) In addition, bactericides, buffers, isotonic agents, surfactants, chelating agents, thickeners, wetting agents, detergency improvers, fragrances and the like can be blended.

  As the disinfectant, although depending on the contact lens type of hard contact lens, oxygen permeable hard contact lens, and soft contact lens, for example, benzethonium chloride, polyhexamethylene biguanide, polydronium chloride are used unless they are contrary to the object of the present invention. Chlorohexidine gluconate, sorbic acid and its salts, thimerosal, chlorobutanol, phenethyl alcohol, p-oxybenzoic acid esters and the like.

  The buffering agent is used so that the pH of the contact lens preparation of the present invention is in the range of about 4 to 10, preferably 5 to 9, more preferably 6 to 8. As a buffering agent, an acid and a salt thereof, and a base and a salt thereof can be used in combination by a conventional method so as to obtain a desired pH. For example, boric acid, borax, citric acid, sodium citrate, tartaric acid, tartaric acid Examples thereof include sodium, gluconic acid, sodium gluconate, acetic acid, sodium acetate, phosphoric acid, sodium monohydrogen phosphate, sodium dihydrogen phosphate, various amino acids, and combinations thereof.

  The isotonic agent is not particularly limited as long as it is water-soluble and does not show adverse effects such as eye irritation. For example, sodium chloride, potassium chloride, calcium chloride, glycerin, sorbitol, glucose, mannitol, propylene glycol and the like can be mentioned.

  As the surfactant, any of a nonionic surfactant, an anionic surfactant, an amphoteric surfactant, and a cationic surfactant can be used. The surfactant is used as a bactericidal agent, a detergency improving agent or the like. Examples of the anionic surfactant include lauroyl sarcosine sodium, lauroyl-L-glutamate triethanolamine, myristyl sarcosine sodium, and the amphoteric surfactants include, for example, lauryl dimethylaminoacetic acid betaine, 2-alkyl-N- Examples of the nonionic surfactant include polysorbate 80, polyoxyethylene hydrogenated castor oil 60, polyoxyl 40 stearate, polyoxyethylene lauryl, and the like, such as carboxymethyl-N-hydroxyethylimidazolinium betaine and alkyldiaminoglycine hydrochloride. Examples of the cationic surfactant include benzethonium chloride, benzalkonium chloride, cetylpyridinium chloride, and the like.

  Examples of the chelating agent include sodium edetate, sodium citrate, condensed sodium phosphate and the like.

  Examples of the thickener include hydroxyethyl cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose sodium, polyethylene glycol, sodium chondroitin sulfate, polyvinyl alcohol, polyvinyl pyrrolidone and the like.

  Examples of the wetting agent include glycerin, polyethylene glycol, propylene glycol and the like.

  Examples of detergency improvers include proteolytic enzymes, lipolytic enzymes, polysaccharide-degrading enzymes, peroxide salts, and surfactants. Examples of proteolytic enzymes include papain, pancreatin, trypsin, bromelain and the like. Examples of the lipolytic enzyme include phospholipase and pancreatic lipase. Examples of polysaccharide-degrading enzymes include chitosan-degrading enzyme, mucin-degrading enzyme, lysozyme, heparinase, and hyaluronidase. Examples of peroxide salts include percarbonate, perborate, and hydrogen peroxide.

  Examples of the fragrances include menthol, camphor, borneol, geraniol and the like. These may be any of d-form, l-form, and dl-form, and may be blended in the form of essential oil.

  Further, when the contact lens composition is a contact lens mounting solution, an eye drop or an eye wash when the contact lens is mounted, in addition to the contact lens adhesion inhibitor of Pseudomonas aeruginosa, a base (for example, distilled water, sterilized purified water, In addition to physiological saline, medicinal ingredients, wetting agents, stabilizers, suspending agents, buffering agents, tonicity agents, chelating agents, thickening agents, and flavoring agents can be blended.

  Here, the medicinal components include vitamins (for example, retinol palmitate, pyridoxine hydrochloride, tocopherol acetate, etc.), vasoconstrictors (for example, naphazoline hydrochloride, tetrahydrozoline hydrochloride, etc.), anti-inflammatory agents (for example, dipotassium glycyrrhizinate, azulene sulfonic acid) Sodium), amino acids (eg, epsilon aminocaproic acid, aminoethylsulfonic acid, aspartic acid or salts thereof), and antiallergic agents such as antihistamines (eg, chlorpheniramine maleate, diphenhydramine hydrochloride, sodium cromoglycate, etc.) Can be mentioned.

  Examples of the solubilizer include polyvinyl pyrrolidone, polyethylene glycol, propylene glycol, polyoxyethylene hydrogenated castor oil 60, polyoxyl 40 stearate, polysorbate 80, and the like. Preservatives include, for example, paraoxybenzoates, benzalkonium chloride, benzethonium chloride, chlorobutanol, benzyl alcohol, sorbic acid or salts thereof, chlorhexidine gluconate, sodium dehydroacetate, cetylpyridinium chloride, alkyldiaminoethylglycine hydrochloride Etc. Examples of the stabilizer include ascorbic acid, sodium edetate, cyclodextrin, condensed phosphoric acid or a salt thereof, sulfite, citric acid or a salt thereof, and the like. Further, examples of the suspending agent include methyl cellulose, hydroxypropyl cellulose, polyvinyl pyrrolidone, polyoxyethylene hydrogenated castor oil 60, polyoxyl 40 stearate, polyethylene glycol, sodium carboxymethyl cellulose, polyvinyl alcohol and the like. Moreover, as a fragrance | flavor, the above-mentioned thing can be illustrated similarly.

  In addition, when using as an eye drop at the time of wearing a contact lens, the dose of the eye drop is preferably administered at a rate of 5 to 6 times a day, for example, in the case of an adult.

  The contact lens composition is not particularly limited as long as it can take a liquid state at the time of use, and examples thereof include a liquid agent and a solid agent that is dissolved at the time of use. Examples of solid agents include tablets, granules, powders, and lyophilized products, but lyophilized products are preferred in view of the speed of dissolution, aseptic conditions, and the uniform surface of the composition. These can be produced by conventional methods. In addition, the compounding amount and pH of the above-mentioned salt of γ-PGA or amino acid-modified γPGA mean the compounding amount and pH when adjusted to a liquid state at the time of use in the case of a solid agent.

  When the composition for contact lenses of the present invention is, for example, a contact lens disinfectant, the contact lens after wearing is placed in this disinfecting solution and used by dipping for 1 to 12 hours. Further, when the contact lens composition of the present invention is, for example, for cleaning and storage of contact lenses, the contact lens after desorption is rubbed with the cleaning preservative and used by immersing in the cleaning preservative for 1 to 12 hours. Is done. The contact lens can be used by rinsing it with water or a contact lens solution applicable to the eyes when the contact lens is mounted.

  The composition for contact lenses of the present invention containing a salt of γ-PGA or amino acid modification-γPGA has an excellent adhesion inhibitory activity against Pseudomonas aeruginosa.

  Next, the present invention will be described in more detail with reference examples, examples and formulation examples. However, the present invention is not limited to these examples. In addition, unless otherwise mentioned in the following, the compounding quantity of each component shall be shown by weight%.

Reference Example 1 Production of γ-polyglutamic acid and its salt (1) γ-polyglutamic acid γ-polyglutamic acid was produced according to the description in JP-B-43-24472.

Specifically, a synthetic medium (pH 7) containing 5% glucose, 0.3% K ₂ HPO ₄ , 0.15% MgSO ₄ , 0.002% MnSO ₄ , and 0.002% FeSO ₄ is prepared. The flask was dispensed and steam sterilized at 120 ° C. for 10 minutes. Separately sterilized urea was added to a concentration of 0.55%. To this, 2% of Bacillus subtilis 5E strain pre-cultured with 0.5% yeast extract was inoculated, and cultured with shaking at 30-31 ° C. for 72 hours. The liquid became viscous as it was cultured. 1 L of the culture solution obtained as described above was collected, dissolved by adding 30 g of NaCl, and 1.5 L of ethanol was added thereto. The precipitated gummy solid was collected, and water was added thereto to prepare a 0.5% strength aqueous solution. Thereto was added NaCl at a rate of 3 g / 100 mL, the pH was adjusted to 9-10 with 6N-NaOH, and the mixture was centrifuged at 14000 rpm for 30 minutes to obtain a supernatant.

  Ethanol was added to the obtained supernatant to 60% by volume to obtain a gum-like substance again. This substance was dissolved in water (0.5% concentration), adjusted to pH 2 or lower by adding 6N hydrochloric acid, filled into a dialysis tube (cut molecular weight: wt. 2000), and purified by dialysis for 2 days as purified water. . The collected dialysate was freeze-dried to obtain γ-polyglutamic acid (about 15 g) having a molecular weight of about 1 million as a white powder.

(2) Sodium salt of γ-polyglutamic acid After adjusting the pH to 10 by adding 10% sodium hydroxide aqueous solution to the above γ-polyglutamic acid, this was filled in a dialysis tube (cut molecular weight: wt.2000) The solution was dialyzed for 2 days as 5 L of purified water. After dialysis, the γ-polyglutamic acid aqueous solution in the tube is lyophilized to prepare a sodium salt of γ-polyglutamic acid.

Example 1 Production of a salt of glycine-modified (γ-polyglutamic acid) (1) Sodium salt of glycine-modified (γ-polyglutamic acid) γ-polyglutamic acid (γ-polyglutamic acid TYPE-H) having a molecular weight of 800,000 to 1,000,000 Manufactured by Nippon Polyglu Co., Ltd .: the same in the following examples) (1294.7 mg), glycine methyl ester hydrochloride (1205.3 mg: 9.6 mmol), and 4-dimethylaminopyridine (97.7 mg: 0.8 mmol) in water 80 mL Was dissolved with stirring. To this was further added 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (1993.7 mg: 10.4 mmol), and the mixture was stirred at 0 ° C. for 2 hours and then at room temperature overnight. A NaOH aqueous solution in which 5.5 g of sodium hydroxide was dissolved in 30 mL of water was added to the obtained solution under cooling (0 ° C.), and the mixture was further stirred overnight at room temperature. Next, 6N aqueous hydrochloric acid was added to make the solution acidic (about pH 5), and the mixture was stirred at room temperature for 1 hour. The solution thus prepared was filled in a dialysis tube (cut molecular weight: wt. 2000), and purified by dialysis for 3 days while changing the external solution every day, using the external solution as purified water (5 L). After dialysis, the content solution was collected and lyophilized to obtain a sodium salt (about 1056 mg) of glycine-modified- (γ-polyglutamic acid) as a white powder (yield: 63.8%). The powder was identified by IR (KBr method) (VALOR-III: manufactured by JASCO Corporation), 1H-NMR (JEOL JNM-EX400 FTNMR SYSTEM) and elemental analysis (N content). Further, from the results of 1H-NMR, the obtained glycine modified- (γ-polyglutamic acid) Na was found to contain glycine in 65% of all α-carboxyl groups of glutamic acid constituting glycine modified- (γ-polyglutamic acid). It was thought to be an amide bond. The glycine-modified- (γ-polyglutamic acid) Na is a sodium salt of amino acid modification- (γ-polyglutamic acid) in which m is 2000 to 2700 and l is 4000 to 5000 in the general formula (I).

IR (KBr method) cm ^-1 : 2940 (-CH _2- ), 3300, 3080, 1647 (CONH), 1598 (CO _2- )
1H-NMR (ppm): 2.01 & 2.13 (1H & 1H, COC H 2 CH 2 CH, m), 3.71 (65% of 2H, NHCH 2, m), 4.19 (1H, COCH 2 CH 2 CHNH, m)
N content: measured value 11.12% (theoretical value 11.99% as 65% modification rate).
Molecular weight: 1 million to 1.3 million.

  (2) In (1) above, glycine-modified (γ-polyglutamic acid) was produced in the same manner except that 4-dimethylaminopyridine (97.7 mg: 0.8 mmol) was not used. As a result, the production yield was 22.5%, which was 41.3% lower than when 4-dimethylaminopyridine was used. From this, it was found that the production yield of glycine-modified- (γ-polyglutamic acid) was increased by using 4-dimethylaminopyridine.

Example 2 Production of Aspartic Acid Modified- (γ-Polyglutamic Acid) and its Salt (1) Sodium Salt of Aspartic Acid-Modified (γ-Polyglutamic Acid) In Example 1 (1), glycine methyl ester hydrochloride (1205.3 mg : 9.6 mmol) in place of L-aspartic acid dimethyl hydrochloride (1897.2 mg: 9.6 mmol), and otherwise treated in the same manner, aspartic acid modification consisting of white powder-(γ-polyglutamic acid) sodium The salt (about 1037 mg) was obtained (yield: 51.4%). The identification result of the powder is shown below. From the results of 1H-NMR, it is considered that the obtained aspartic acid-modified- (γ-polyglutamic acid) Na has aspartic acid amide-bonded to 57% of the total α-carboxyl group of glutamic acid constituting it. It was. The aspartic acid modification- (γ-polyglutamic acid) Na is a sodium salt of amino acid modification- (γ-polyglutamic acid) in which m is 2600 to 3300 and l is 3500 to 4100 in the general formula (I).

IR (KBr method) cm ^-1 : 2945 (-CH _2- ), 3300, 3078, 1646 (CONH), 1600 (CO _2- )
1H-NMR (ppm): 1.88 & 2.02 (1H & 1H, COCH 2 C H 2 CH, m), 2.30 (2H, COC H 2 CH 2 CH, m), 2.73 (57% of 2H, CH 2 CO 2 H , m), 4.10 (1H, COCH ₂ CH ₂ CHNH, m), 4.45 (57% of 1H, NH-CH-)
N content: measured value 9.58% (theoretical value 9.53% as a modification rate of 57%).
Molecular weight: 1.2 million to 1.6 million.

(2) Aspartic acid modification-(γ-polyglutamic acid) carboxylic acid type Furthermore, the sodium salt obtained in (1) is dissolved in water, and the pH of the solution is acidic (approximately 2 or less) using 6N-HCl. And stirred overnight. Subsequently, this was filled in a dialysis tube (cut molecular weight: wt. 2000), and purified by dialysis for 3 days while changing the external solution every day, using the external solution as purified water (5 L). After dialysis, the content solution was collected and freeze-dried to obtain a carboxylic acid form (about 765 mg) of aspartic acid-modified (γ-polyglutamic acid) as a white powder (yield: 45.9%). The identification result of the powder is shown below.

IR (KBr method) cm ^-1 : 2945 (-CH _2- ), 3300, 3075, 1647 (CONH), 1729 (CO _2- )
1H-NMR (ppm): 1.96 & 2.21 (1H & 1H, COCH 2 C H 2 CH, m & m), 2.40 (2H, COC H 2 CH 2 CH, m), 2.82 (57% of 2H, CH 2 CO 2 H , m), 4.28 (1H, COCH ₂ CH ₂ CHNH, m), 4.65 (57% of 1H, NH-CH-)
N content: measured value 11.03% (theoretical value 11.21% as 57% modification rate)
Molecular weight: 1.2 million to 1.6 million.

  (3) Aspartic acid-modified (γ-polyglutamic acid) Na was produced in the same manner as in (1) above except that 4-dimethylaminopyridine (97.7 mg: 0.8 mmol) was not used. As a result, the production yield was 20.7%, which was 25.2% lower than when 4-dimethylaminopyridine was used. From this, it was confirmed that the production yield of aspartic acid-modified (γ-polyglutamic acid) increases by using 4-dimethylaminopyridine.

Example 3 Production of Salt of Glutamic Acid Modification-(γ-Polyglutamic Acid) (1) Glutamic Acid Modification-Sodium Salt of (γ-Polyglutamic Acid) In Example 1 (1), glycine methyl ester hydrochloride (1205.3 mg: 9.6 mmol) ) Instead of L-glutamic acid diethyl hydrochloride (2301.1 mg: 9.6 mmol), otherwise obtain the sodium salt of glutamic acid-modified (γ-polyglutamic acid) (about 1137 mg) consisting of white powder (Yield: 53.4%). The identification result of the powder is shown below. From the results of 1H-NMR, it was considered that the obtained glutamic acid-modified (γ-polyglutamic acid) Na had glutamic acid amide-bonded to 60% of the total α-carboxyl group of glutamic acid constituting it. . The glutamic acid modification- (γ-polyglutamic acid) Na is a sodium salt of amino acid modification- (γ-polyglutamic acid) in which m is 2400 to 3100 and l is 3600 to 4700 in the general formula (I).

IR (KBr method) cm ^-1 : 2943 (-CH _2- ), 3300, 3075, 1650 (CONH), 1600 (CO _2- )
1H-NMR (ppm): 1.88 & 2.05 (1H & 1H, COCH ₂ CH ₂ CH, m & m), 2.13 (60% of 2H, COCH ₂ CH ₂ CO ₂
H, m), 2.31 (60% of 4H, COCH ₂ CH ₂ CH, CH ₂ CO ₂ H, m), 4.14 (60% of 1H, NH-CH-, m), 4.23 (1H, COCH ₂ CH ₂ CHNH, m)
N content: measured value 9.20% (theoretical value 9.27% as 60% modification rate)
Molecular weight: 1.4 million to 1.8 million.

  (2) Glutamic acid-modified (γ-polyglutamic acid) Na was produced in the same manner as in (1) except that 4-dimethylaminopyridine (97.7 mg: 0.8 mmol) was not used. As a result, the production yield was 19.8%, which was 33.6% lower than when 4-dimethylaminopyridine was used. From this, it was found that the production yield of glutamic acid-modified (γ-polyglutamic acid) was increased by using 4-dimethylaminopyridine.

Example 4 Production of a salt of serine-modified (γ-polyglutamic acid) (1) Sodium salt of serine-modified (γ-polyglutamic acid) In Example 1 (1), glycine methyl ester hydrochloride (1205.3 mg: 9.6 mmol) In the same manner as above, serine methyl hydrochloride (1493.6 mg: 9.6 mmol) was used, and the sodium salt of serine-modified (γ-polyglutamic acid) (about 1219 mg) consisting of white powder was obtained in the same manner. Rate: 68.1%). The identification result of the powder is shown below. From the result of 1H-NMR, it was considered that the obtained serine-modified- (γ-polyglutamic acid) Na had serine amide-bonded to 60% of the total α-carboxyl group of glutamic acid constituting it. . The serine-modified- (γ-polyglutamic acid) Na is a sodium salt of amino acid modification- (γ-polyglutamic acid) in which m is 2400 to 3100 and l is 3600 to 4700 in the general formula (I).

IR (KBr method) cm ^-1 : 2944 (-CH _2- ), 3300, 3080, 1645 (CONH), 1597 (CO _2- )
1H-NMR (ppm): 1.86 & 2.04 (1H & 1H, COCH ₂ CH ₂ CH, m & m), 2.28 (2H, COCH ₂ CH ₂ C
H, m), 3.78 (60 % of 2H, CH 2 OH, m), 4.08 (60% of 1H, NHCH-, m), 4.23 (1H, COCH 2 CH 2 CHNH, m)
N content: measured value 10.14% (60% modification rate, theoretical value 10.77%)
Molecular weight: 1.2 million to 1.5 million.

  (2) Serine-modified (γ-polyglutamic acid) Na was produced in the same manner as in (1) except that 4-dimethylaminopyridine (97.7 mg: 0.8 mmol) was not used. As a result, the production yield was 20.3%, which was 47.8% lower than when 4-dimethylaminopyridine was used. From this, it was found that the production yield of serine-modified (γ-polyglutamic acid) was increased by using 4-dimethylaminopyridine.

Experimental Example 1 Moisturizing Test Using the keratin powder, the water retention ability of the amino acid-modified (γ-polyglutamic acid) produced in Examples 1 to 4 or a salt thereof was measured by the following method.
<Measurement of water retention ability>
(1) Prepare a 5% aqueous test solution using the sample listed in Table 1 as the test sample.
(2) 1 g of keratin powder (manufactured by Tokyo Chemical Industry Co., Ltd.) is put into a beaker having a diameter of 2.5 cm, and 200 μL of the 5% test aqueous solution prepared in (1) is added dropwise thereto.
(3) Put this in a 35 ° C incubator, measure the weight after 1 hour, and calculate the amount of water evaporation from the difference from the weight before placing in the incubator.

  The results are shown in Table 1. In the table, the relative water evaporation rate (%) indicates the water evaporation rate (%) when the water evaporation amount is 100% when water is used as the test sample. In Table 1, γ-polyglutamic acid used in Comparative Example 3 as a test sample is γ-polyglutamic acid (γ-polyglutamic acid TYPE-H, γ-polyglutamic acid TYPE-H having a molecular weight of 800,000 to 1,000,000 used as a starting material in Examples 1-4. Lipidure (registered trademark) Lipidure PMB (ph10) (manufactured by Nippon Oil & Fats Co., Ltd.) used in Comparative Example 1 is a phosphatidylcholine that constitutes a cell membrane. It is a water-soluble polymer having 2-methacryloyloxyethyl phosphorylcholine and butyl methacrylate having the same structure as the polar group, and is known to have moisture retention. In addition, acetyl hyaluronic acid (manufactured by Shiseido) used in Comparative Example 2 is a product also referred to as super hyaluronic acid because it has twice the moisturizing power of hyaluronic acid.

  As can be seen from this result, although γ-polyglutamic acid has almost the same water retention ability (moisturizing action) as acetyl hyaluronic acid having twice the moisturizing power of hyaluronic acid, it is modified with glycine- (γ-polyglutamic acid). The salt of Example 1 (Example 1), aspartic acid modification- (γ-polyglutamic acid) and its salt (Example 2) all had a water retention capacity (moisturizing effect) superior to that of γ-glutamic acid. . Further, the salt of glutamic acid modification- (γ-polyglutamic acid) (Example 3) and the salt of serine modification- (γ-polyglutamic acid) (Example 4) are inferior to γ-polyglutamic acid but have moisture retention. It had a moisture retention capacity (moisturizing effect) superior to Lipidure PMB (ph10). This shows that the amino acid modification- (γ-polyglutamic acid) and salt of the present invention have excellent moisturizing properties.

Experimental example 2 Staphylococcus aureus adhesion inhibitory action (1) Staphylococcus aureus adhesion inhibitory action About the amino acid modification-((gamma) -polyglutamic acid) or its salt manufactured in Examples 1-4 by the following method with respect to epidermis The inhibitory effect on S. aureus adhesion inhibition was evaluated.

(1) Dissolve the sample listed in Table 2 as a test sample in sterilized purified water to prepare a 2% test aqueous solution. In Table 2, γ-polyglutamic acid Na is γ-polyglutamic acid having a molecular weight of 800,000 to 1,000,000 (γ-polyglutamic acid TYPE-H, manufactured by Nippon Polyglu Co., Ltd.) described in Reference Example 1 (2). (Molecular weight: 900,000 to 1.1 million) (the same applies to (2)).
(2) Using a glass slide with double-sided tape attached, keratinocytes are collected from the upper arm of women in their 30s.
(3) 500 μL of the 2% aqueous test solution prepared in (1) is dropped on the keratinocytes of the slide glass and dried in a clean bench for 20 minutes.
(4) Apply 20 μL of 10 ⁷ CFU / MmL bacterial solution to Staphylococcus aureus ATCC6538 and dry for 20 minutes.
(5) Wash 5 times with 1mL physiological saline.
(6) The site where Staphylococcus aureus is applied is collected by the contact plate method (microbe detection medium sheet “Sani thick microbial detection medium sheet”, manufactured by Chisso Corporation), and the number of Staphylococcus aureus is measured after culture. .
(7) S. aureus adhesion inhibition rate (%) of each test sample is compared with the number of S. aureus when phosphate buffered saline [PBS (-)] is used as a test sample (control example). Then, the number of reductions is obtained, and the inhibition rate relative to the control example (S. aureus adhesion inhibition rate 0%) is calculated from the number of reductions.

  The results are shown in Table 2.

  As can be seen from this result, the salt of γ-polyglutamic acid and its amino acid modified product [amino acid modification— (γ-polyglutamic acid) salt] markedly suppressed the adhesion of Staphylococcus aureus to the skin.

(2) Adhesion inhibitory action against Staphylococcus epidermidis (skin resident bacteria) In (1) above, Staphylococcus epidirmidis ATCC12228 is used in place of Staphylococcus aureus ATCC6538. The inhibitory action of each test sample on the adhesion of Staphylococcus epidermidis to the skin was evaluated. In addition, Staphylococcus epidermidis is a skin resident bacteria that normally exists on the surface of healthy skin, and keeps the skin weakly acidic while decomposing sebum together with acne bacteria, and prevents the invasion of bad bacteria from the outside. have. In order to maintain healthy skin, it is important that staphylococcus epidermidis maintain a superior skin condition.

  The results are shown in Table 3.

  As can be seen from the results, the salt of γ-polyglutamic acid and the amino acid modified form thereof [amino acid modification— (γ-polyglutamic acid) salt] are different from Staphylococcus epidermidis, and are resident skin bacterium (good bacteria). Adhesion inhibitory action against is weak, and it is effective for maintaining healthy skin by controlling adhesion to the skin according to the type of bacteria.

Experimental Example 3 Pseudomonas aeruginosa Adhesion Inhibiting Action on Contact Lenses According to the following method, Pseudomonas aeruginosa adhesion inhibitory action on contact lenses was obtained for the amino acid modification- (γ-polyglutamic acid) or salt thereof produced in Examples 1-2. evaluated.
(1) Dissolve the samples listed in Table 4 as test samples in sterilized purified water, and prepare a test aqueous solution of 0.1% concentration at pH 7.5 using 4N hydrochloric acid or sodium hydroxide aqueous solution.
(2) About 100 mL of the test aqueous solution is filled in a polyethylene container, and a soft contact lens (2-week Accuview Group IV, manufactured by Johnson & Johnson) is placed in the container and immersed for 2 weeks. Replace the test aqueous solution with a fresh one every other week.
(3) After 2 weeks, the contact lens is taken out of the test aqueous solution, and immersed in 5 mL of 10 ⁸ CFU / MmL bacterial solution of Pseudomonas aeruginosa ATCC9027 for 1 minute.
(4) After 1 minute, the lens is taken out from the bacterial solution, placed in physiological saline (5 mL), shaken lightly for 1 minute by hand, then replaced in fresh physiological saline (5 mL), and ultrasound (ultrasonic) for 3 minutes. Sonic cleaner, Sono Cleaner 200R Kaijo).
(5) After sonication, the number of bacteria in the physiological saline is measured by the SCDLP agar smear method (n = 3).

  The results are shown in Table 4.

  As can be seen from this result, γ-polyglutamic acid and its amino acid modification [amino acid modification— (γ-polyglutamic acid) salt] markedly suppressed adhesion of Pseudomonas aeruginosa to the contact lens.

Formulation Example 1 Skin Cream (pH 6)
Aspartic acid modification- (γPGA) Na (Example 2) 2.0 (%)
White petrolatum 10.0
Stearyl alcohol 2.0
Glyceryl monostearate 1.0
Polyoxyethylene (40) hydrogenated castor oil 2.0
Glucerin 5.0
1,3-butylene glycol 10.0
Xanthan gum 0.4
Lactic acid or sodium lactate appropriate amount Methylparaben 0.1
Purified water balance
Total 100.0%.

Formulation Example 2 Lotion (pH 5)
Glutamic acid modification- (γPGA) Na (Example 3) 2.0 (%)
1,3-butylene glycol 10.0
Glucerin 5.0
Polyoxyethylene (40) hydrogenated castor oil 0.5
Methylparaben 0.05
Lactic acid or sodium lactate
Purified water balance
Total 100.0%.

Formulation Example 3 Eye drops for use with contact lenses (pH 7)
Aspartic acid modification- (γPGA) Na (Example 2) 2.0 (%)
Polysorbate 80 0.05
l-Menthol 0.005
Hydroxyethyl cellulose 0.07
Sodium chloride 0.5
Boric acid 1.0
Borax 0.2
pH adjuster (hydrochloric acid or sodium hydroxide) Adjust to pH 7
Purified water balance
Total 100.0%.

Formulation Example 4 Contact Lens Preservation Cleaning Solution (pH 7.2)
Aspartic acid modification- (γPGA) Na (Example 2) 0.02 (%)
Polyoxyethylene (40) hydrogenated castor oil 0.02
Hydroxypropyl methylcellulose 0.1
Polyhexamethylene biguanide 0.0001
Sodium chloride 0.7
Sodium hydrogen phosphate 0.2
Sodium dihydrogen phosphate 0.015
pH adjuster (hydrochloric acid or sodium hydroxide) Adjust to pH 7.2
Purified water balance
Total 100.0%.

Claims (12)

  Amino acid modification (γ-polyglutamic acid) or a salt thereof, in which a part or all of the side chain α-carboxyl group of γ-polyglutamic acid and an amino group of a hydrophilic amino acid are amide-bonded. Formula (I)

[In the formula, R is a hydrophilic group represented by the following formula (II),

(In the formula (II), R ₁ represents a lower alkylene group, R ₂ represents a lower alkylene group which may be substituted with a hydroxyl group, and R ₃ represents a hydrogen atom, a hydroxyl group or a carboxyl group. X and y represent , The same or different, indicating 0 or 1.)
m represents an integer of 0 or 1 or more, l represents an integer of 6 to 15000 together with m, and each repeating unit is present randomly or partially forms a block. ]
The amino acid modification- (γ-polyglutamic acid) or a salt thereof according to claim 1, which is represented by:   The amino acid modification-(γ) according to claim 2, wherein the hydrophilic group (II) is a group obtained by removing an amino group from at least one hydrophilic amino acid selected from the group consisting of glycine, serine, aspartic acid, and glutamic acid. -Polyglutamic acid) or a salt thereof.   The amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of claims 1 to 3, having a molecular weight of 1,000 to 5,000,000.   5. The method according to claim 1, further comprising reacting γ-polyglutamic acid with an alkyl ester of a hydrophilic amino acid in the presence of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride. A process for producing amino acid modification- (γ-polyglutamic acid) or a salt thereof.   γ-polyglutamic acid and an alkyl ester of a hydrophilic amino acid are reacted in the presence of 4-dimethylaminopyridine in addition to 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride, A process for producing the amino acid modification- (γ-polyglutamic acid) or a salt thereof according to claim 5.   A moisturizer comprising the amino acid modification (γ-polyglutamic acid) according to any one of claims 1 to 4 or a salt thereof as an active ingredient.   A staphylococcus aureus comprising as an active ingredient at least one compound selected from the group consisting of γ-polyglutamic acid, amino acid modification according to any one of claims 1 to 4 (γ-polyglutamic acid), and salts thereof Or an adhesion inhibitor of Pseudomonas aeruginosa.   A skin external preparation containing the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of claims 1 to 4.   Cosmetics containing the amino acid modification- (γ-polyglutamic acid) or a salt thereof according to any one of claims 1 to 4.   A hair care agent comprising the amino acid modification (γ-polyglutamic acid) or a salt thereof according to any one of claims 1 to 4.   A composition for contact lenses, comprising at least one compound selected from the group consisting of γ-polyglutamic acid, amino acid modification according to any one of claims 1 to 4 (γ-polyglutamic acid), and salts thereof. .