JP2018162440A - Composition and use therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a composition containing modified polysaccharides, which has reduced hardening shrinkage and can form a gel with excellent mechanical strength.SOLUTION: A composition contains a modified polysaccharide (A) having a hydroxy aryl group or an unsaturated double bond-containing group as a crosslinkable group, a crosslink promoter (B) for the modified polysaccharide (A), and a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof.SELECTED DRAWING: None

Description

  The present invention relates to compositions and uses thereof.

  When a part of living tissue such as cartilage and bone is damaged, replacement with an artificial medical instrument using a hydrogel is performed at the damaged part. As such hydrogels, the following polymer networks (gels) are known. In other words, it is a polymer network (gel) formed by oxidative coupling of a modified polysaccharide in which a hydroxyphenyl group is introduced into a polysaccharide using an enzyme such as horseradish peroxidase and hydrogen peroxide as a catalyst. (See Patent Documents 1 to 4).

U.S. Pat. No. 8,287,906 US Pat. No. 6,982,298 US Published Patent No. 2012/301441 US Pat. No. 9,132,201

  A gel formed from a modified polysaccharide in which a hydroxyphenyl group is introduced into the polysaccharide has a small curing shrinkage upon gelation. For this reason, when the gel is used as an artificial medical instrument for replacement of a living tissue, the gel is very easy to handle. However, according to the study of the present inventors, the gel is brittle and the mechanical strength is not sufficient.

  An object of the present invention is to provide a composition capable of forming a gel having a small curing shrinkage during gelation and excellent mechanical strength. Moreover, it aims at providing the medical device which has the gel formed from the said composition, the molded object formed from the said gel, and the said molded object. Furthermore, it aims at providing the manufacturing method and solution kit of a gel.

The present inventor has intensively studied to solve the above problems. As a result, it has been found that a composition having the following constitution can solve the above problems, and has completed the present invention. The present invention relates to the following [1] to [15], for example.
[1] The crosslinkable group is selected from a modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group, a crosslinking accelerator (B) of the modified polysaccharide (A), a carboxyl group and a salt thereof. And a polysaccharide (C) having at least one group.
[2] The modified polysaccharide (A) is a modified polysaccharide (A1) having a hydroxyaryl group, and the crosslinking accelerator (B) is a combination of an enzyme (B1) and a peroxide (B2). ] The composition of description.
[3] The modified polysaccharide (A1) having the hydroxyaryl group is at least one selected from phenolic hydroxyl group-modified hyaluronic acid, phenolic hydroxyl group-modified dextran, and phenolic hydroxyl group-modified pullulan. Composition.
[4] The composition according to any one of [1] to [3], wherein the polysaccharide (C) is at least one selected from alginic acid, pectin, and derivatives thereof.
[5] The composition according to any one of [1] to [4], further containing water (E).
[6] The composition according to any one of [1] to [5], further including a metal salt (D).
[7] The composition according to [6], wherein the metal salt (D) is a Group 2 metal salt of the periodic table.
[8] A gel having a structure in which a first network structure and a second network structure exist in an intertwined state, and the first network structure has hydroxyaryl as a crosslinkable group. A network structure formed by crosslinking a modified polysaccharide (A) having a group or an unsaturated double bond-containing group, wherein the second network structure is at least one selected from a carboxyl group and a salt thereof. A gel characterized by a network structure formed by a polysaccharide (C) having a group and a metal cation (D ').
[9] The gel according to [8], which is a hydrogel containing 50 to 99.9% by mass of water.
[10] A molded product comprising 50% by mass or more of the gel according to [8] or [9].
[11] A medical instrument comprising the molded article according to [10].
[12] A step of applying the composition according to any one of [1] to [5] to a damaged part of a biological tissue to gel the composition, and the obtained gel and metal cation (D A method for producing an interpenetrating network-structured gel, comprising: bringing the gel into contact with an aqueous solution containing ').
[13] The composition according to any one of [1] to [5] is gelled, the obtained gel is brought into contact with an aqueous solution containing a metal cation (D ′), and the gel is cured. Interpenetrating network type gel formed by letting
[14] Modified polysaccharide (A) having a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, and having a hydroxyaryl group or an unsaturated double bond-containing group, and the modified polysaccharide (A) A first solution containing any one of the crosslinking accelerators (B) but not both, and one of the modified polysaccharide (A) and the crosslinking accelerator (B) not included in the first solution. A solution kit comprising: a second solution; and a third solution containing a metal salt (D).
[15] A modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group and a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, and an enzyme (B1) And the eleventh solution containing either of the peroxide (B2) but not both, and the enzyme (B1) and the peroxide (B2) that are not included in the eleventh solution A solution kit comprising a twenty-first solution and a thirty-first solution containing a metal salt (D).

  ADVANTAGE OF THE INVENTION According to this invention, the composition which can form the gel which has small cure shrinkage at the time of gelatinization, and was excellent in mechanical strength, the gel formed from the said composition, and the molded object formed from the said gel And a medical device having the molded body, and further a method for producing a gel and a solution kit can be provided.

1 (1), (2) and (3) are photographs of hydrogels of Example 1, Comparative Example 1 and Comparative Example 2, respectively.

Hereinafter, modes for carrying out the present invention will be described.
Each chemical substance exemplified in the present specification, for example, a chemical substance contained in the composition or used in each step can be used alone or in combination of two or more unless otherwise specified. Can be used.

[Composition]
The composition of the present invention comprises a modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group (hereinafter collectively referred to as “crosslinkable group (a1)”) as the crosslinkable group, It contains a crosslinking accelerator (B) of the modified polysaccharide (A) and a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, preferably further containing a metal salt (D). .

  In the composition containing the modified polysaccharide (A), the crosslinking accelerator (B) and the polysaccharide (C), the modified polysaccharide (A) is crosslinked with the modified polysaccharide (A) using the crosslinking accelerator (B). A first network structure formed by crosslinking is obtained. At this stage, a gel (semi-interpenetrating polymer network gel) in which the polysaccharide (C) is entangled with the first network structure can be formed. It is considered that the first network structure and the polysaccharide (C) do not have chemical bonds such as covalent bonds and exist in a state where they are intertwined with each other by physical bonds such as intermolecular forces.

Further, by bringing the semi-interpenetrating polymer network gel into contact with the metal cation (D ′) of the metal salt (D), a second network structure formed by crosslinking the polysaccharide (C) is obtained. It is done. At this stage, a gel (interpenetrating polymer network gel) having a structure in which the first network structure and the second network structure exist in an intertwined state can be formed. The first network structure and the second network structure do not have a chemical bond such as a covalent bond that binds to each other, and are entangled with each other by a physical bond such as an intermolecular force. It is thought that it exists in the state.
Details of the gel structure will be described later.

  The polysaccharide (C) has a property of being crosslinked by a metal cation such as a calcium cation. For example, a gel obtained from alginic acid and a calcium cation has high mechanical strength, but is hard to shrink during gelation, and the resulting gel is difficult to use as a medical instrument such as artificial cartilage. On the other hand, the gel formed from the modified polysaccharide (A) having a crosslinkable group (a1) has small curing shrinkage upon gelation.

  For this reason, the interpenetrating polymer network gel (IPN gel) described above has an excellent effect that it has a high mechanical strength and also has a small curing shrinkage during gelation during the production of the gel. For example, when a plurality of types of gels are mixed, the respective advantages are often canceled out, but the IPN gel of the present invention is a gel that exhibits an excellent effect without canceling out the respective advantages.

<Modified polysaccharide (A)>
The modified polysaccharide (A) is usually a compound in which a crosslinkable group (a1), that is, a hydroxyaryl group or an unsaturated double bond-containing group is introduced into the polysaccharide (a).
Examples of the polysaccharide (a) constituting the modified polysaccharide (A) include hyaluronic acid, alginic acid, xanthan gum, cellulose, guar gum, pullulan, dextran, fructan, mannan, carrageenan, chitin, chitosan, pectin, starch, glycogen, dextrin, Examples include gellan gum and derivatives thereof. Among these, at least one selected from hyaluronic acid, dextran, and pullulan is preferable.

  Examples of the derivative of the polysaccharide (a) include various modified products, and the site to be modified and the modifying group (excluding the crosslinkable group (a1)) should not significantly impair the effects of the present invention. There is no particular limitation. For example, a derivative in which the exemplified polysaccharide (a) has a substituent such as methyl group, ethyl group, hydroxyethyl group, hydroxypropyl group, hydroxyethylmethyl group, hydroxypropylmethyl group, carboxymethyl group, acetyl group, stearoxy group Is mentioned. The derivative may have the substituent alone or in combination.

  Specific examples of the polysaccharide (a) having the substituent include acetylated hyaluronic acid, propylene glycol hyaluronate, propylene glycol alginate, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl methyl cellulose, Examples thereof include carboxymethyl cellulose, stearoxyhydroxypropyl methylcellulose, methyl guar gum, ethyl guar gum, hydroxyethyl guar gum, hydroxypropyl guar gum, hydroxyethyl methyl guar gum, and hydroxypropyl methyl guar gum.

  The polysaccharide (a) may be a salt, and examples thereof include acid addition salts, metal salts, and ammonium salts. Examples of the acid addition salt include inorganic acid salts such as hydrochloride, sulfate, and phosphate, and organic acid salts such as acetate, maleate, fumarate, tartrate, and citrate. Examples of the metal salt include alkali metal salts such as sodium salt and potassium salt. For example, the metal salt of the polysaccharide which has a carboxyl group is mentioned. The polysaccharide having a carboxyl group may be an ester thereof.

  The modified polysaccharide (A) has a hydroxyaryl group or an unsaturated double bond-containing group. Examples of the hydroxyaryl group include a hydroxyphenyl group, and examples of the unsaturated double bond-containing group include a (meth) acryloyl group, which is preferable because it has a smaller curing shrinkage during gelation. Is a hydroxyaryl group.

  The crosslinkable group (a1) is determined depending on the combination with the crosslinking accelerator (B). Examples of the combination include a hydroxyaryl group such as a hydroxyphenyl group, an enzyme (B1), and a peroxide (B2 if necessary). And a combination of an unsaturated double bond-containing group such as a (meth) acryloyl group and a radical polymerization initiator (B3). Among these, a combination of a hydroxyaryl group such as a hydroxyphenyl group, an enzyme (B1), and, if necessary, a peroxide (B2) is preferable.

  Carbon number of a hydroxyaryl group is 6-50 normally, Preferably it is 6-30. In the hydroxyaryl group, the number of hydroxyl groups bonded to the benzene nucleus is usually 1 or more, preferably 1 to 3, more preferably 1 to 2. Thus, a hydroxyaryl group is not limited to a monohydroxyaryl group, and similarly, a hydroxyphenyl group is not limited to a monohydroxyphenyl group.

  Examples of the hydroxyaryl group include a hydroxyphenyl group, a hydroxynaphthyl group, and a hydroxyanthracenyl group. The hydroxyaryl group may have a substituent such as an alkyl group. In the hydroxyaryl group, it is preferable that a hydrogen atom is bonded to a carbon atom in at least one ortho position with respect to the hydroxyl group.

  The unsaturated double bond in the unsaturated double bond-containing group is preferably a carbon-carbon double bond. As an unsaturated double bond containing group, a (meth) acryloyl group and a vinyl group are mentioned, for example, A (meth) acryloyl group is preferable and an acryloyl group is more preferable.

  The modified polysaccharide (A) is preferably a compound in which a hydroxyaryl group is bonded to the polysaccharide (a) by an ester bond, a urethane bond or an amide bond, directly or via an organic group. The ester bond and the urethane bond are preferably bonded to the carbon atom to which the hydroxyl group or carboxyl group that can be contained in the polysaccharide (a) is bonded, and the amide bond is a carboxyl group that can be contained in the polysaccharide (a). It is preferably bonded to the bonded carbon atom. The carboxyl group may be a salt or an ester.

  Examples of the organic group include, for example, an alkanediyl group having 1 to 20 carbon atoms, and a substitution in which at least one selected from an ether bond and a divalent amino group (—NH— or the like) is added to or inserted into the alkanediyl group. An alkanediyl group, an organic group having a peptide bond, specifically, a group derived from an oligopeptide such as a dipeptide or an ester thereof.

  As a method for introducing a hydroxyaryl group into the polysaccharide (a), for example, a polysaccharide (a) having a functional group such as an amino group, an alcoholic hydroxyl group and a carboxyl group, and a functional group capable of reacting with the functional group (for example: And a method of reacting a compound having a carboxyl group, an acid halide thereof, an amino group, an alcoholic hydroxyl group, an epoxy group, an oxetanyl group) and a hydroxyaryl group.

  Examples of the compound having a carboxyl group capable of reacting with an alcoholic hydroxyl group in the polysaccharide (a) or an acid halide thereof and a hydroxyaryl group include 4-hydroxyphenylacetic acid, 4-hydroxyphenylpropionic acid, 4-hydroxy Examples include phenylbutanoic acid, 2- (2,5-dihydroxyphenyl) acetic acid, and acid halides thereof.

  When the polysaccharide (a) has a carboxyl group, its salt or ester, examples of the compound having an amino group capable of reacting with the group and a hydroxyaryl group include tyramine.

  The modified polysaccharide (A) is preferably a phenolic hydroxyl group-modified polysaccharide (A1), more preferably at least one selected from phenolic hydroxyl group-modified hyaluronic acid, phenolic hydroxyl group-modified dextran, and phenolic hydroxyl group-modified pullulan. Specifically, a modified polysaccharide in which a hydroxyphenyl group is introduced into the polysaccharide (a) is preferable, and at least one selected from hydroxyphenyl group-modified hyaluronic acid, hydroxyphenyl group-modified dextran, and hydroxyphenyl group-modified pullulan is more preferable. .

Hereinafter, one embodiment of the modified polysaccharide (A) will be described.
For example, after converting an alcoholic hydroxyl group possessed by a large amount of polysaccharide (a) to a carbonate ester derivative group using a carbonylating reagent, the resulting carbonate ester derivative of polysaccharide (a) and an amine compound having a hydroxyaryl group And react.

  Examples of the carbonylation reagent include nitrophenyl chloroformate, phenyl chloroformate, trichloromethyl chloroformate, and bis (trichloromethyl) carbonate. The amount of the carbonylating reagent is usually 1 to 100 parts by mass with respect to 100 parts by mass of the polysaccharide (a). When the carbonic acid ester derivative that is an intermediate is stable, the following reaction can be carried out after it is once isolated.

  Examples of the amine compound having a hydroxyaryl group include tyramine. The amount of the amine compound is usually 1 to 100 parts by mass with respect to 100 parts by mass of the carbonate ester derivative of the polysaccharide (a).

  The reaction is preferably performed in the presence of a basic compound. Examples of the basic compound include trimethylamine, triethylamine, tributylamine, pyridine, and piperidine.

  The reaction is preferably performed in a solvent. Examples of the solvent include amide solvents such as dimethylformamide and dimethylacetamide; sulfoxide solvents such as dimethyl sulfoxide; halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane and chloroform; aromatics such as benzene, toluene and xylene. Hydrocarbon; ethers such as diethyl ether, tetrahydrofuran, dioxane and the like can be mentioned. In order to improve the solubility of the polysaccharide (a) in the solvent, an alkali metal salt such as lithium chloride can be used.

  In each step, the reaction temperature is usually −10 to 50 ° C., preferably −5 to 30 ° C., and the reaction time is usually 0.1 to 24 hours.

  As a method for introducing an unsaturated double bond-containing group into the polysaccharide (a), for example, the polysaccharide (a) having a functional group such as an amino group, an alcoholic hydroxyl group and a carboxyl group can react with the functional group. A functional group (eg, carboxyl group, its acid halide, amino group, alcoholic hydroxyl group, epoxy group, oxetanyl group) and a compound having an unsaturated double bond (hereinafter also referred to as “double bond-containing compound”), The method of making it react is mentioned.

  Examples of the double bond-containing compound include a carboxyl group capable of reacting with an alcoholic hydroxyl group contained in a large amount of the polysaccharide (a) or an acid halide thereof (a group represented by —COX (X is a halogen atom such as a chlorine atom)). For example, unsaturated monocarboxylic acids such as (meth) acrylic acid and crotonic acid; unsaturated dicarboxylic acids such as maleic acid and fumaric acid; anhydrides of the unsaturated dicarboxylic acids; succinic acid mono [2- (Meth) acryloyloxyethyl] and mono [(meth) acryloyloxyalkyl] esters of polyvalent carboxylic acids such as mono [2- (meth) acryloyloxyethyl] phthalate; and acid halides thereof such as chloride ( (Meth) acryloyl; and the like.

  Other examples of the double bond-containing compound include epoxy group-containing (meth) acrylates such as glycidyl (meth) acrylate and 3,4-epoxycyclohexylmethyl (meth) acrylate; o, m or p-vinylbenzylglycidyl Epoxy group-containing vinyl compounds such as ether, o, m or p-isopropenylbenzylglycidyl ether; Oxetanyl group-containing (meth) acrylates such as 3-((meth) acryloyloxymethyl) oxetane; 3- (p-vinylphenyloxy Examples include oxetanyl group-containing vinyl compounds such as methyl) -3-methyloxetane; and hydroxyalkyl (meth) acrylates such as 2-hydroxyethyl (meth) acrylate.

  In the reaction, the double bond-containing compound is usually used in an amount of 1 to 500 parts by mass, preferably 5 to 300 parts by mass, more preferably 10 to 100 parts by mass with respect to 100 parts by mass of the polysaccharide (a). be able to.

  The reaction is preferably performed in the presence of a basic compound. Examples of the basic compound include trimethylamine, triethylamine, tributylamine, pyridine, and piperidine.

  The reaction is preferably performed in a solvent. Examples of the solvent include water, methanol, ethanol, isopropyl alcohol, dimethylformamide, dimethyl sulfoxide, and dimethylacetamide.

The reaction temperature of the reaction is usually −10 to 50 ° C., preferably −5 to 30 ° C., and the reaction time is usually 0.1 to 24 hours.
The modified polysaccharide (A) is preferably a modified polysaccharide in which a (meth) acryloyl group is introduced into the polysaccharide (a) ((meth) acryloyl modified polysaccharide), (meth) acryloyl modified hyaluronic acid, (meth) acryloyl modified dextran, and At least one selected from (meth) acryloyl-modified pullulan is more preferable.

The degree of substitution (DS) with the crosslinkable group (a1) in the modified polysaccharide (A) is usually 0.1 to 300, preferably 1 to 100, more preferably 2 to 30. The degree of substitution can be determined from ¹ H NMR. The degree of substitution of the modified polysaccharide (A) indicates the average number of crosslinkable groups (a1) introduced per 100 units of the constituent monosaccharide of the modified polysaccharide. Since the modified polysaccharide has such a substitution degree, the gelation rate of the modified polysaccharide is improved.

  The number average molecular weight of the modified polysaccharide (A) by the absolute molecular weight measurement using a multi-angle light scattering detector is usually 1,000 to 10,000,000, preferably 3,000 to 1,000,000, more preferably. Is 5,000 to 500,000. Moreover, the molecular weight distribution (weight average molecular weight / number average molecular weight; both are based on the absolute molecular weight measurement) of the modified polysaccharide (A) is usually 1 to 20, preferably 1 to 10, and more preferably 1 to 5.

One or more modified polysaccharides (A) can be used.
The content rate of modified polysaccharide (A) in the composition of this invention is 0.05-49 mass% normally, Preferably it is 0.5-30 mass%, More preferably, it is 3-15 mass%. Such an embodiment is preferable from the viewpoints of the mechanical strength and handleability of the gel.

<Crosslinking accelerator (B)>
The crosslinking accelerator (B) is used for promoting the crosslinking of the crosslinking group (a1) of the modified polysaccharide (A).
Examples of the crosslinking accelerator (B) when the modified polysaccharide (A) has a hydroxyaryl group as the crosslinkable group (a1) include, for example, an enzyme (B1) that directly or indirectly promotes crosslinking of the hydroxyaryl group. Is mentioned.

Examples of the enzyme (B1) include laccase, tyrosinase, catalase, and peroxidase. Laccase and tyrosinase are excellent in that there is no possibility that the function-expressing substance is denatured during gelation because there is no need to add other chemicals that do not constitute gel components. Peroxidase can be instantly gelled by using it together with a peroxide such as hydrogen peroxide. Examples of the origins of copper enzymes such as laccase and tyrosinase (phenol oxidase) include urushi, mushrooms (Tuchikaburi, mushrooms), and molds (Polyporus vericolor). The origins of hydroperoxidases such as catalase and peroxidase are, for example, bovine liver, horse blood cells, human blood cells, M. lysodeikticus, horseradish (horseradish), soybean, radish, turnip, thyroid, milk, intestine, white blood cells, red blood cells, Examples include yeast, Caldariomyces fumago and Steptococcus faecalis. Among these, tyrosinase is preferably derived from mushrooms and peroxidase is preferably derived from horseradish. As the enzyme (B1), peroxidase and catalase are preferable, and peroxidase is more preferable.
One or more enzymes (B1) can be used.

  In the composition of the present invention, the amount of the enzyme (B1) is usually from 0.01 to 10000 U, preferably from 0.1 to 2000 U, more preferably from 1 mol of the hydroxyaryl group in the modified polysaccharide (A). 1 to 1000 U. In one embodiment, the amount of enzyme (B1) in the composition of the present invention is preferably 0.01 to 10 U / mL, more preferably 0.1 to 5 U / mL from the viewpoint of reactivity. . U represents a unit of enzyme activity, and represents the amount of enzyme capable of changing 1 micromole of substrate per minute at a temperature of 30 ° C. under optimum conditions.

  When the enzyme (B1) is used as the crosslinking accelerator (B), particularly when peroxidase and / or catalase is used, it is preferable to further use a peroxide (B2). That is, a combination of enzyme (B1) and peroxide (B2) is preferable.

Examples of the peroxide (B2) include hydrogen peroxide.
In the composition of the present invention, the amount of the peroxide (B2) is usually 0.01 to 1000 mol, preferably 0.1 to 500 mol, per 1 mol of the hydroxyaryl group in the modified polysaccharide (A). More preferably, it is 0.5-200 mol. In one embodiment, the amount of peroxide (B2) in the composition of the present invention is preferably 0.1 to 10 mM, more preferably 0.5 to 5 mM, from the viewpoint of reactivity.

  Examples of the crosslinking accelerator (B) when the modified polysaccharide (A) has an unsaturated double bond-containing group as the crosslinkable group (a1) include a radical polymerization initiator (B3). Examples of the radical polymerization initiator (B3) include photo radical polymerization initiators such as alkylphenone compounds, acylphosphine oxide compounds, biimidazole compounds, triazine compounds, oxime ester compounds, benzoin compounds, and benzophenone compounds; Examples thereof include thermal radical polymerization initiators such as azo compounds.

One or more radical polymerization initiators (B3) can be used.
In the composition of the present invention, the amount of the radical polymerization initiator (B3) is usually 1 to 100 parts by mass with respect to 100 parts by mass of the modified polysaccharide (A).

<Polysaccharide (C)>
The polysaccharide (C) is a polysaccharide having at least one group selected from a carboxyl group and a salt thereof. For example, at least one selected from alginic acid, pectin and derivatives thereof is preferable. However, the modified polysaccharide (A) described above is excluded from (C). Examples of the carboxyl group salt include alkali metal salts such as sodium salt and potassium salt, and ammonium salt.

  Examples of derivatives of alginic acid and pectin include salts and esters. Examples of the salt include acid addition salts, metal salts, and ammonium salts. Examples of the acid addition salt include inorganic acid salts such as hydrochloride, sulfate, and phosphate, and organic acid salts such as acetate, maleate, fumarate, tartrate, and citrate. Examples of the metal salt include alkali metal salts such as sodium salt and potassium salt. Examples of the ester include alkyl esters, alkenyl esters, alkynyl esters, aryl esters, arylalkyl esters, alkanoyloxyalkyl esters, and alkoxycarbonyloxyalkyl esters. Metal salts, ammonium salts, and esters are, for example, the aforementioned salts and esters of the carboxyl group possessed by alginic acid and pectin.

The polysaccharide (C) preferably does not have the same crosslinkable group as that of the modified polysaccharide (A), for example, a hydroxyaryl group, from the viewpoint of forming an interpenetrating polymer network gel.
The number average molecular weight of the polysaccharide (C) by the absolute molecular weight measurement using a multi-angle light scattering detector is usually 10,000 to 10,000,000, preferably 20,000 to 5,000,000, more preferably 50,000 to 2,000,000. The molecular weight distribution of the polysaccharide (C) (weight average molecular weight / number average molecular weight; both based on the absolute molecular weight measurement) is usually 1 to 20, preferably 1.5 to 10, and more preferably 2 to 8. .
One or more polysaccharides (C) can be used.

  The content of the polysaccharide (C) in the composition of the present invention is usually 0.01 to 2 g, preferably 0.05 to 1 g, more preferably 0.1 to 0, based on 1 g of the modified polysaccharide (A). .4 g. Such an embodiment is preferable from the viewpoints of the mechanical strength and handleability of the gel.

<Metal salt (D)>
The composition of the present invention preferably further contains a metal salt (D). However, the aforementioned modified polysaccharide (A) and polysaccharide (C) are excluded from the metal salt (D). The metal salt (D) acts as a crosslinking agent for the polysaccharide (C).

The metal salt (D) is preferably a polyvalent metal salt, more preferably a Group 2 metal salt of the periodic table, and even more preferably an alkaline earth metal salt. Examples of the polyvalent metal cation include divalent metal cations such as Ca ²⁺ , Mg ²⁺ , Ba ²⁺ , Sr ²⁺ , and Cu ²⁺ , and trivalent metal cations such as Al ³⁺ and Fe ^3+. It is done. The metal salt (D) is preferably a polyvalent metal halide, sulfate or nitrate, and examples thereof include CaCl ₂ , MgCl ₂ , CaSO ₄ , BaCl ₂ , AlCl ₃ , and Fe (NO ₃ ) ₃ .
One or more metal salts (D) can be used.

  The content ratio of the metal salt (D) or the polyvalent metal cation in the composition of the present invention is usually 0.01 to 1000 mol, preferably 1 mol in total for the carboxyl group of the polysaccharide (C) and the salt thereof. Is 0.1 to 100 mol, more preferably 1 to 50 mol. Such an embodiment is preferable from the viewpoints of the mechanical strength and handleability of the gel.

<Water (E)>
The composition of the present invention preferably contains water (E).
Content of water (E) in the composition of this invention is 50-99.9 mass% normally, Preferably it is 65-99 mass%, More preferably, it is 80-96 mass%. In such an embodiment, the gel tends to be excellent in mechanical strength and handleability.

<Other ingredients>
The composition of this invention may contain the function expression substance according to the use of the gel. Thereby, a function expression substance can be included in the gel obtained. Examples of the function-expressing substance include drugs, cells, and cell growth factors. Gels containing a function-expressing substance can be used in a wide range of fields such as medical instruments.

  The composition of the present invention may further contain a polysaccharide other than the modified polysaccharide (A) and the polysaccharide (C) described above. As this polysaccharide, the polysaccharide (a) described in the column of <modified polysaccharide (A)> is mentioned, for example.

<Aspect of composition>
One embodiment of the composition of the present invention is a multi-part solution kit.
One aspect of the solution kit of the present invention includes a first solution containing a polysaccharide (C) and containing either a modified polysaccharide (A) or a crosslinking accelerator (B) but not both, and the above-mentioned modified polysaccharide ( It has the 2nd solution containing the one which is not contained in a 1st solution among A) and a crosslinking accelerator (B), and a 3rd solution containing a metal salt (D). Each of the first to third solutions is preferably an aqueous solution.

  Alternatively, an aspect of the solution kit of the present invention includes an eleventh solution that includes the modified polysaccharide (A) and the polysaccharide (C), and includes either the enzyme (B1) or the peroxide (B2) but not both. And a twenty-first solution containing the enzyme (B1) and peroxide (B2) not included in the eleventh solution and a thirty-first solution containing the metal salt (D). Each of the 11th to 31st solutions is preferably an aqueous solution.

  Each solution is mixed, for example, with a microtube or a triple syringe described in JP-A-2014-501570, for example, immediately before gel formation or treatment.

  The composition of this invention can be used suitably as a raw material composition of the gel demonstrated below. In addition, the composition of the present invention can be used, for example, as a lubricant in a joint cavity used for the treatment of osteoarthritis or a damage covering material. Osteoarthritis can occur in body joints such as knees, shoulders, hips, hips, ankles, wrists, and fingers, but the composition of the present invention can be applied to any joint.

[gel]
The gel of the present invention is a gel having a structure in which the first network structure and the second network structure are intertwined with each other, so-called interpenetrating polymer network gel (IPN gel). It is.

The network structure means, for example, a cross-linked structure obtained by cross-linking polymers and having a three-dimensional network structure.
The first network structure is a network structure formed by crosslinking a modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group as a crosslinkable group. In one embodiment, when the hydroxyaryl group of the modified polysaccharide (A) is reacted with the peroxide (B2) in the presence of the enzyme (B1), a hydrogen atom is extracted at the phenolic hydroxyl group to form a free radical species, Isomerized into free radical species with unpaired electrons on the carbon at the ortho position, then two free radical species are combined and then enolized to form a network (crosslinked structure). It is thought.

  The second network structure is a network structure formed by a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, and a metal cation (D ′). The polysaccharide (C) has a property of being crosslinked by a metal cation such as a calcium cation. Therefore, it is considered that a network structure (crosslinked structure) is formed by the polysaccharide (C) and the metal cation (D ′). The metal cation (D ′) is preferably a metal cation derived from the metal salt (D) described above.

  The IPN gel is excellent in handleability since it has a small cure shrinkage when forming the IPN gel including the first network structure and the second network structure. The IPN has a high mechanical strength. For this reason, it is possible to easily produce a medical instrument made of the IPN gel having a shape corresponding to the affected area of the patient, a high fixation rate to the affected area, and excellent mechanical strength.

The shape of the gel of the present invention is not particularly limited, and is formed into a shape conforming to the shape to which the gel is applied, such as a columnar shape, a cylindrical shape, a disc shape, a spherical shape, a square shape, an elliptical shape, a fiber shape, and a rod shape. can do.
In one embodiment, the gel is a hydrogel usually containing 50 to 99.9% by mass, preferably 70 to 96% by mass. Hydrogels retain water in interpenetrating polymer networks. The hydrogel generally contains the first network structure (polymer) and the second network structure (polymer) in a total amount of 50 to 0.1% by mass, preferably 30 to 4% by mass. . The hydrogel is excellent in mechanical strength and flexibility.

The hydrogel of the present invention has, for example, a large compression modulus, breaking stress and breaking strain, and is excellent in mechanical strength. These physical properties are values calculated by the measurement methods described in the examples.
When the hydrogel of the present invention is used for artificial cartilage, its hardening shrinkage is preferably 10% or less. The cure shrinkage rate is a value calculated by the measurement method described in the examples.

The hydrogel of this invention can be obtained with the manufacturing method mentioned later. Moreover, a xerogel can be obtained by removing the water | moisture content by drying the said hydrogel suitably.
The gel and hydrogel of the present invention can be suitably used as, for example, artificial cartilage and artificial bone when a part of living tissue such as cartilage and bone is damaged.

[Gel production method]
The interpenetrating network structure type gel of the present invention includes, for example, a modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group as a crosslinkable group, and a crosslinking accelerator (B) of the modified polysaccharide (A). And a step of gelling a composition containing a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof (hereinafter also referred to as “gelation step”), and the obtained gel It can be obtained by a production method comprising a step of bringing the gel into contact with an aqueous solution containing a metal cation (D ′) (hereinafter also referred to as “curing step”).

<Gelification process>
The gelation step is a step of forming the first network structure. In the presence of the polysaccharide (C), the polysaccharide (C) is entangled with the first network structure by crosslinking the modified polysaccharide (A) having the crosslinkable group (a1) with the crosslinking accelerator (B). A semi-interpenetrating polymer network gel is formed.

  For example, the first solution containing the polysaccharide (C) and containing either the modified polysaccharide (A) and the crosslinking accelerator (B), but not both, and the modified polysaccharide (A) and the crosslinking accelerator (B) Mixing with a second solution containing one that is not contained in the first solution; alternatively, containing modified polysaccharide (A) and polysaccharide (C), and containing enzyme (B1) and peroxide (B2) An eleventh solution containing either but not both is mixed with the twenty-first solution containing the enzyme (B1) and peroxide (B2) that are not included in the eleventh solution. Then, the modified polysaccharide (A) is crosslinked to obtain a semi-interpenetrating polymer network gel. The first, second, eleventh and twenty-first solutions are preferably all aqueous solutions.

  In the gelation step, for example, the composition is applied to a damaged part of a living tissue to gel the composition. Examples of damaged parts of living tissue include cartilage and bone defects. Alternatively, in the gelation step, a mold having a cavity having a desired shape is prepared, and each component of the composition is mixed and reacted in the cavity to cause gelation having the shape.

  The temperature at which the gel is formed is usually 4 to 50 ° C., preferably 10 to 45 ° C., more preferably 20 to 40 ° C., and the time is usually 0.01 to 10 hours, preferably 0. 1 to 5 hours.

<Curing process>
The curing step is a step of forming the second network structure. The polysaccharide (C) is considered to be entangled with the first network structure, and in this state, the polysaccharide (C) is cross-linked with the metal cation (D ′), so-called interpenetrating polymer network structure. A mold gel (IPN gel) can be formed.

  As the metal cation (D ′), a polyvalent metal cation is preferable. The aqueous solution containing a metal cation (D ′) can be obtained, for example, by mixing a metal salt (D) and water (E).

  In the curing step, for example, the gel obtained in the gelation step is brought into contact with the third or thirty-first solution containing the metal salt (D), for example, the gel is immersed in the third or thirty-first solution. Then, the gel is cured. The third or thirty-first solution is preferably an aqueous solution. Content of the metal salt (D) in the 3rd or 31st solution is 1-1000 mM normally, Preferably it is 10-500 mM.

  The temperature at which the gel in the curing step is immersed in the third or 31st solution is usually 4 to 70 ° C., preferably 10 to 50 ° C., and the time is usually 1 to 100 hours, preferably 5 to 50 hours.

[Molded body and medical device]
The molded object of this invention contains the gel mentioned above. For example, the gel of the present invention is a molded body formed into a desired shape. In the molded body, the amount of gel is usually 50% by mass or more, preferably 60% by mass or more, and more preferably 70% by mass or more.

  The medical instrument of this invention has the said molded object. Since the said medical instrument has the said molded object which is excellent in mechanical strength and a softness | flexibility, it can be used suitably for an implant material especially among medical instruments. Examples of the implant material include an implant material used when a part of biological tissue such as cartilage and bone is damaged.

EXAMPLES Hereinafter, although this invention is demonstrated further more concretely based on an Example, this invention is not limited to these Examples.
<Production of composition and hydrogel>
[Synthesis Example] Dextran Modified with Hydroxyphenyl Group (A1-1)
To the flask were added 40 g of dextran (trade name “Dextran 40”, manufactured by Meisei Kogyo Co., Ltd.), 1600 ml of dimethylformamide, and 30.9 g of lithium chloride, and the mixture was stirred at 90 ° C. for 90 minutes in a nitrogen atmosphere. The stirred solution was ice-cooled, 9.2 ml of pyridine and 23.8 g of nitrophenyl chloroformate were added, and the mixture was stirred at 0 ° C. for 60 minutes. 2000 ml of cold ethanol was added to the stirred solution, reprecipitation was performed, and the precipitate was filtered and washed to obtain a crude product.

  The crude product and 740 ml of dimethylformamide were added to the flask, and the mixture was stirred at 23 ° C. for 30 minutes under a nitrogen atmosphere, 9.1 g of tyramine was added, and the mixture was stirred at 23 ° C. for 180 minutes. 800 ml of cold ethanol was added to the stirred solution, reprecipitation was performed, the precipitate was filtered, dissolved in pure water, impurities were removed by dialysis, and dextran (A1-1) modified with a hydroxyphenyl group An aqueous solution was obtained.

When the substitution degree (DS) of the obtained dextran (A1-1) modified with a hydroxyphenyl group was calculated by ¹ H NMR, DS = 14.
_{^{1 H NMR (400MHz, D 2}} O): δ 2.75 and 3.05 (m, -CH 2 -CH 2 -), 3.3-4.1 (m, dextran glucosidic protons), 4.2-5.6 (s, dextran anomeric protons), and 6.86 and 7.17 (d, tyramine aromatic protons).

[Preparation Examples 1 to 10] Modified polysaccharide (A) and polysaccharide (C) aqueous solution (polysaccharide aqueous solution)
The aqueous solution of dextran (A1-1) modified with the hydroxyphenyl group and the alginic acid aqueous solution shown in Table 1 below are mixed so that the ratio of dextran (A1-1) and alginic acid is shown in Table 1 below. Then, it was prepared to be a polysaccharide aqueous solution having the concentrations shown in Table 1 below with pure water.
The details of the alginic acid aqueous solution are as follows.
IL-2: Sodium alginate manufactured by Kimika Co., Ltd., trade name “IL-2”
IL-6: Sodium alginate manufactured by Kimika Co., Ltd., trade name “IL-6”
IL-6G: Kimika Co., Ltd. sodium alginate, trade name “IL-6G”
IL-6M: Kimika Co., Ltd. sodium alginate, trade name “IL-6M”
ULV-20: Sodium alginate manufactured by Kimika Co., Ltd., trade name “ULV-20”

[Examples 1 to 8] Production of compositions and hydrogels The aqueous polysaccharide solution, the horseradish peroxidase aqueous solution (concentration: 150 U / ml), and pure water prepared in Preparation Examples 1 to 8 with the contents shown in Table 2 below. Place in a microtube (tube having a cylindrical hole with a diameter of 17 mm) and mix evenly. Then, stirring with a vortex mixer, hydrogen peroxide solution (concentration: 88.2 mM) is added to the mixture at a normal temperature. The mixture was stirred for 30 minutes to form a semi-interpenetrating polymer network gel (semi-IPN gel).

Subsequently, an aqueous calcium chloride solution (concentration: 100 mM) was poured into the microtube, and the semi-IPN gel was immersed at 23 ° C. for 12 hours to produce a hydrogel (interpenetrating polymer network gel). The hydrogel was cylindrical and transparent.
FIG. 1 (1) is a photograph of the hydrogel of Example 1.

[Comparative Example 1] Production of Composition and Hydrogel 600 μl of the aqueous polysaccharide solution prepared in Preparation Example 9, 6.66 μl of horseradish peroxidase aqueous solution (concentration: 150 U / ml), and 359 μl of pure water were provided in a microtube (column shape having a diameter of 17 mm). In a tube having a hole of 2), and while stirring with a vortex mixer, 34 μl of hydrogen peroxide (concentration: 88.2 mM) is added to the mixed solution at a time and stirred at room temperature for 30 minutes. A gel (containing 92.5% by mass of water) was produced. The hydrogel was cylindrical and transparent.
FIG. 1 (2) is a photograph of the hydrogel of Comparative Example 1.

Comparative Example 2 Production of Composition and Hydrogel 600 μl of the aqueous polysaccharide solution prepared in Preparation Example 10 was placed in a microtube (tube having a cylindrical hole with a diameter of 17 mm), and then an aqueous calcium chloride solution (concentration: 100 mM). Poured to produce a hydrogel. The hydrogel was cylindrical, wrinkled on the surface, and white.
FIG. 1 (3) is a photograph of the hydrogel of Comparative Example 2.

<Evaluation>
[Curing shrinkage]
In the production of the hydrogels of Examples 1 to 8 and Comparative Examples 1 and 2, the curing shrinkage rate in gelation was calculated by the following formula. The evaluation results are shown in Table 3 below.
[(Diameter of microtube hole−diameter of hydrogel) / diameter of microtube hole] × 100 (%)

[Mechanical strength]
The hydrogels of Examples 1 to 8 and Comparative Examples 1 to 2 were cut into a cylindrical shape having a diameter of 8 mm and a thickness of 6 mm by using an egg slicer to prepare an evaluation gel. A compression elastic modulus, breaking stress, and breaking strain were measured using an evaluation gel, an Instron (trade name “33R4204”, manufactured by Instron) and a load cell. The compression modulus was calculated from a 10-15% stress-strain curve. The compression was performed at 0.6 mm / min (constant displacement) and the load was 50N. The evaluation results are shown in Table 3 below.

Claims (15)

A modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group as a crosslinkable group;
A crosslinking accelerator (B) of the modified polysaccharide (A);
A composition comprising a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof.   The modified polysaccharide (A) is a modified polysaccharide (A1) having a hydroxyaryl group, and the crosslinking accelerator (B) is a combination of an enzyme (B1) and a peroxide (B2). Composition.   The composition according to claim 2, wherein the modified polysaccharide (A1) having a hydroxyaryl group is at least one selected from phenolic hydroxyl group-modified hyaluronic acid, phenolic hydroxyl group-modified dextran, and phenolic hydroxyl group-modified pullulan.   The composition according to any one of claims 1 to 3, wherein the polysaccharide (C) is at least one selected from alginic acid, pectin, and derivatives thereof.   The composition according to any one of claims 1 to 4, further comprising water (E).   The composition according to any one of claims 1 to 5, further comprising a metal salt (D).   The composition according to claim 6, wherein the metal salt (D) is a Group 2 metal salt of the periodic table. A gel having a structure in which the first network structure and the second network structure exist in an intertwined state;
The first network structure is a network structure formed by crosslinking a modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group as a crosslinkable group;
The second network structure is a network structure formed by a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, and a metal cation (D ′). Gel to do.   The gel according to claim 8, which is a hydrogel containing 50 to 99.9% by mass of water.   A molded body comprising the gel according to claim 8 or 9 in an amount of 50% by mass or more.   A medical instrument comprising the molded article according to claim 10.   Applying the composition according to any one of claims 1 to 5 to a damaged part of a living tissue to gel the composition; an aqueous solution containing the obtained gel and a metal cation (D '); A method for producing an interpenetrating network-structured gel, comprising: bringing the gel into contact with each other.   It formed by making the composition of any one of Claims 1-5 gelatinize, contacting the obtained gel and the aqueous solution containing a metal cation (D '), and hardening the said gel. Interpenetrating network structure gel. Crosslinking promotion of modified polysaccharide (A) having polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, having a hydroxyaryl group or an unsaturated double bond-containing group, and the modified polysaccharide (A) A first solution comprising either agent (B) but not both;
A second solution containing one of the modified polysaccharide (A) and the crosslinking accelerator (B) not included in the first solution;
A third solution containing a metal salt (D);
A solution kit comprising: A modified polysaccharide (A) having a hydroxyaryl group or an unsaturated double bond-containing group, and a polysaccharide (C) having at least one group selected from a carboxyl group and a salt thereof, an enzyme (B1) and peroxidation An eleventh solution containing either of the products (B2) but not both;
A twenty-first solution containing the enzyme (B1) and the peroxide (B2) not included in the eleventh solution;
A thirty-first solution containing a metal salt (D);
A solution kit comprising: