Gel-forming material and gel composition

Abstract

The present invention pertains to a gel-forming material comprising: a component A which is at least one substance selected from the group consisting of modified hyaluronic acids having an amino group and an average molecular weight of 10,000 or more, and salts thereof; and genipin.

Description

Technical Field

[0001] The present invention relates to a gel-forming material and a gel composition. Background Art

[0002] Materials that gel in situ can be used as medical materials that can be injected into the body, and therefore, they can achieve minimally invasive treatment and are attracting attention in the fields of drug carriers and tissue engineering. For materials that gel in situ, in addition to gelling in a relatively short time in response to changes in temperature, pH, and chemical substances, biocompatibility and safety are also important issues. Among various gel-forming materials, hyaluronic acid has excellent biocompatibility and safety to the human body. As a composition for obtaining a hydrogel using hyaluronic acid, for example, Patent Document 1 discloses a hyaluronic acid derivative having a specific structure and its cross-linked product as a composition that can be used for the treatment of cytokine-related inflammation. [Prior art literature] [Patent Document]

[0003] Patent Document 1: Japanese Patent Publication No. 2016-518464 Summary of the invention [Technical problem to be solved by the invention]

[0004] The present invention provides a gel-forming material and a gel composition using hyaluronic acid that can gel in a short time at 37° C. The gel-forming material and the gel composition are suitable for administering medical materials that need to gel in situ into the body by injection.

[0005] An object of one embodiment of the present invention is to provide a gel-forming material that can form a gel in a short time under the condition of 37° C. An object of one embodiment of the present invention is to provide a novel in situ gel-forming material. [Technical solutions for solving technical problems]

[0006] The inventors of the present invention have discovered a gel-forming material comprising: component A, which is at least one selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof; and genipin, and the gel-forming material can solve the above-mentioned technical problems. The present invention was completed based on this discovery.

[0007] That is, the present invention relates to, for example, the following technical aspects. [1] A gel-forming material comprising: at least one component A selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof; and genipin. [2] The gel-forming material according to [1], wherein the amino group modification rate of the component A is 5% or more and 70% or less. [3] The gel-forming material according to [1] or [2], wherein the average molecular weight of the component A is 10,000 or more and 1,500,000 or less. [4] A material for in situ gel formation, comprising at least one component selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more, and salts thereof. [5] A material for in-situ gel formation, comprising genipin. [6] A gel composition comprises at least one component A cross-linked by genipin, wherein the component A is selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof. [Effects of the invention]

[0008] According to the present invention, it is possible to provide a gel-forming material that can form a gel in a short time under the condition of 37° C. According to the present invention, it is also possible to provide a novel in situ gel-forming material. DETAILED DESCRIPTION

[0009] Hereinafter, the mode for implementing the present invention will be described in detail. However, the present invention is not limited to the following embodiment.

[0010] <Features of the Invention> (Gel-forming materials) The present invention is characterized by providing a gel-forming material, comprising at least one component A selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof; and genipin. The gel-forming material of the present invention can gel in a short time of less than 1 hour at 37°C and can be applied to medical materials that require in-situ gelation.

[0011] <Gel-forming materials> The "gel-forming material" in this specification is a material containing one or more components that can form a gel composition by reacting with other components. The gel-forming material of the present invention can form a gel composition within 1 hour from the start of the reaction when reacting with other components at 37°C. The gel-forming material of the present invention comprises: at least one component A selected from the group consisting of modified hyaluronic acid introduced with an amino group and salts thereof; and genipin.

[0012] <In situ gel formation> The gel-forming material of the present invention can form a gel in a short time of less than 1 hour at 37° C., and is therefore suitable as an in situ gel-forming material. An “in situ gel-forming material” is a gel-forming material that can form a gel at a target location. The gel-forming material of the present invention can be sterilized in a state before gel formation (e.g., in a solution state). Therefore, sterilization is not necessarily required in a gelled state, and the sterilized gel-forming material can be administered to a target site in the body to form a gel.

[0013] <Ingredient A> Component A is a compound or a salt thereof that introduces a functional group having an amino group (-NH2) into at least a portion of hyaluronic acid. Among them, the acetyl group of N-acetylglucosamine of a portion of hyaluronic acid is deacetylated to become an amino group. The gel-forming material may contain one or more components A.

[0014] (Structure 1 of ingredient A) Component A may be a compound or a salt thereof in which at least a part of the functional groups contained in a part of the disaccharide units in the repeating unit constituting the hyaluronic acid is substituted with a functional group having an amino group. Component A may be a compound or a salt thereof in which at least one group selected from the group consisting of the hydroxyl group at the C4 position and the hydroxyl group at the C6 position of N-acetylglucosamine constituting the hyaluronic acid, and the hydroxyl group at the C2 position, the hydroxyl group at the C3 position, and the -OH group at the C6 position of the glucuronic acid constituting the hyaluronic acid is substituted with a functional group having an amino group.

[0015] (Structure 2 of Component A) Component A may be, for example, a compound represented by the following formula (1) or a salt thereof. [Chemical formula 1] In formula (1), R 1 represents -OH or a functional group having an amino group, R 2 ~R 5 Each of them independently represents a hydrogen atom or a functional group having an amino group. n represents a number of 1 to 7500.

[0016] (Number and Substitution Position of Functional Groups Having Amino Groups in Formula (1)) In formula (1), R 1 ~R 5At least one of them is a functional group having an amino group. In formula (1), R 1 ~R 5 1 to 4, 1 to 3 or 1 to 2 in the above formula is a functional group having an amino group, or R 1 ~R 5 Any one of them is a functional group having an amino group. Alternatively, in formula (1), R 1 is a functional group having an amino group and R 2 ~R 5 They are all hydrogen atoms.

[0017] (Structure 3 of ingredient A) Component A may be, for example, a compound represented by the following formula (1a) or a salt thereof. [Chemical formula 2] In formula (1a), R 1 and n are respectively the same as R in formula (1) 1 Has the same meaning as n.

[0018] (Structure having an amino functional group) Examples of the amino group include an amino group bonded to a hydrocarbon group and an amino group in a hydrazide group. The functional group having an amino group may be a functional group represented by the formula (I): -X 1 -R-NH2 represented by the group.

[0019] (Structure 1 of R in formula (I)) R represents a divalent group, which is 1 A group connected to NH2. The divalent group may include a hydrocarbon group. The hydrocarbon group may be linear or branched. The hydrocarbon group may be, for example, an alkylene group. The number of carbon atoms of the hydrocarbon group and the alkylene group may be, for example, 1 or more or 2 or more, or may be 10 or less, 8 or less, 6 or less, 4 or less, or 3 or less. R may include a functional group other than a hydrocarbon group. The functional group other than a hydrocarbon group may be, for example, -CO-NH-.

[0020] (Structure 2 of R in formula (I)) R can be -R b -R a -R c -represented by a group. a represents an alkylene group, R b and R c Each independently represents a single bond or *1-CO-NH-*2. Here, *1 represents the same a The bonding site of *2 indicates the bonding site with X 1or amino group. R may include an alkylene group with 2 or more carbon atoms. When R includes an alkylene group with 2 or more carbon atoms, the elasticity of the obtained gel composition is further improved. When R includes an alkylene group with 4 or less carbon atoms, the solubility in solvents such as water or PBS is further improved.

[0021] (X in formula (I) 1 Structure) X 1 At the bonding site is R 1 In the case of, it may be -NH- or -O-. 1 At the bonding site is R 2 ~R 5 In the case of , it may be -(C=O)- or a single bond.

[0022] (Specific examples of functional groups having an amino group) The functional group with amino group is R 1 In the case of, it may be -NH-CH2-CH2-NH2, -NH-NH-CO-CH2-CH2-CH2-CH2-CO-NH-NH2, -NH-CH2-NH2, -NH-CH2-CH2-CH2-NH2 or -NH-CH2-CH2-CH2-CH2-NH2. The functional group having an amino group is R at the bonding site. 2 ~R 5 In the case of, it may be -O-CH2-CH2-NH2, -(C=O)-CH2-NH2 or -CH2-CH2-NH2.

[0023] (Salt of modified hyaluronic acid) The salt of the modified hyaluronic acid may be a pharmaceutically acceptable salt. Examples of the salt of the modified hyaluronic acid include alkali metal salts such as sodium salts and potassium salts, and ammonium salts.

[0024] (Preferred upper limit of average molecular weight of component A) The average molecular weight of component A may be 1,500,000 or less, 1,400,000 or less, 1,300,000 or less, 1,200,000 or less, 1,100,000 or less, or 1,000,000 or less. When the average molecular weight of component A is 1,000,000 or less, solubility in solvents such as water and PBS is further improved.

[0025] (Preferred lower limit of average molecular weight of component A) The average molecular weight of component A may be 10000 or more, 20000 or more, 30000 or more, 40000 or more, 50000 or more, 60000 or more, 70000 or more, 80000 or more, 90000 or more, or 100000 or more. When the average molecular weight of component A is 10000 or more, the elasticity of the obtained gel composition is further improved.

[0026] (Preferred numerical range of average molecular weight of component A) The average molecular weight of Component A is preferably 10,000 to 1,500,000 or 100,000 to 1,000,000, and more preferably 100,000 to 800,000, from the viewpoint of further improving solubility in solvents such as water and PBS.

[0027] (Method for determining average molecular weight) The average molecular weight can be measured by the following method. First, using gel filtration column, a plurality of (purification) hyaluronic acids (reference material) with known molecular weight are carried out to liquid chromatography, thus a calibration curve is made according to their retention time. Similarly, the component A as the assay object is carried out to liquid chromatography, the calibration curve made is used to obtain molecular weight, thus the molecular weight of component A can be obtained.

[0028] (Measurement apparatus and measurement conditions of average molecular weight) The 2690 separation module manufactured by Waters Corporation of Japan was used as a liquid chromatography analysis device. The 996 photodiode array manufactured by Waters Corporation of Japan was used as a photodiode array. A column formed by connecting 1 TSK guard column PWXL (manufactured by Tosoh Corporation) and 2 TSK gel GMPW (manufactured by Tosoh Corporation) in series in the order of recording was used as a column. In the determination of the average molecular weight, the conditions of column temperature 40°C, measurement wavelength 210nm, flow rate 0.8mL / min, sample injection volume 20μL, analysis time 40 minutes, and mobile phase 0.003mol / L phosphate buffer-0.15mol / LNaCl (pH7.0) were used. The details of other experimental conditions can be as described in the embodiments described later.

[0029] (Definition of amino modification rate) The amino modification rate of component A refers to the number of amino groups contained in each of the two sugar units constituting hyaluronic acid as one unit, specifically, the ratio (%) of the number of amino groups contained in each of the two sugar units constituting hyaluronic acid to the one unit when the one unit is set to 100%. In this specification, "two sugar units constituting hyaluronic acid" refers to one unit consisting of two sugars (glucuronic acid and N-acetylglucosamine) that are adjacently bonded to constitute hyaluronic acid.

[0030] (Determination method of amino modification rate) Amino modification rate can be used 1 The measurement was performed by H-NMR. Specific examples of the method for preparing the sample for measurement and the measurement method are as described in the examples described later.

[0031] (Lower limit of amino group modification rate) The amino modification rate may be 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, or 10% or more. When the amino modification rate is 10% or more, the gel forming ability is further improved even if the concentration of component A in the gel forming material is a low concentration of less than 1%. From the viewpoint of being able to form a gel in a shorter time, the amino modification rate may be 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, or 55% or more.

[0032] (Preferred upper limit of amino group modification rate) The amino modification rate may be 90% or less, 85% or less, 80% or less, 75% or less, or 70% or less. When the amino modification rate is 70% or less, the solubility in solvents such as water and PBS is further improved.

[0033] (Preferred numerical range of amino group modification rate) From the viewpoint of further improving the solubility in a solvent such as water or PBS, the amino group modification rate is preferably 5% or more and 70% or less, and more preferably 10% or more and 60% or less.

[0034] (Form of Component A) In the gel-forming material, component A may be in liquid or solid form (e.g., powder form). The gel-forming material may include a solution containing component A and a solvent or a powder containing component A. Examples of the solvent include water and a buffer solution. Examples of the buffer solution include a phosphate buffer solution and the like.

[0035] (Solution containing component A) In the solution containing component A, the content of component A can be 0.1 mg or more, 0.5 mg or more, 1 mg or more, greater than 1 mg, 2 mg or more, 3 mg or more, 4 mg or more, or 5 mg or more, and can be 25 mg or less, 15 mg or less, or 10 mg or less, relative to 1 mL of the solvent.

[0036] (Method for producing component A) Component A can be obtained, for example, by a method comprising a reaction step, in which hyaluronic acid and a raw material compound are reacted in a reaction solution containing hyaluronic acid and a group capable of reacting with a functional group in hyaluronic acid and an amino group. The method for manufacturing component A can also include a post-treatment step, in which the reactant obtained by the reaction step is post-treated.

[0037] (Raw material compound) The starting compound can be of formula (Ia): X 2 -R-NH2 or a salt thereof. R in formula (Ia) may be the same group as R in formula (I). In formula (Ia), X 2 Represents a group that can react with the functional group in hyaluronic acid. As a group that can react with the functional group in hyaluronic acid, for example, amino (-NH2) can be listed. The usage amount of the raw material compound can be appropriately adjusted according to the amino modification rate of the target component A, etc.

[0038] (Specific examples of raw material compounds) Examples of the raw material compound include ethylenediamine, adipic acid dihydrazide, methylenediamine, 1,3-propylenediamine (1,3-diaminopropane), 1,4-butylenediamine, 2-aminoethanol, glycine, and 2-chloroethyldiamine.

[0039] (Solvent in the reaction process) The reaction solution may contain a solvent. Examples of the solvent include water or a mixed solution of water and a water-soluble organic solvent such as ethanol.

[0040] (Reactants in the reaction process) The reaction solution may contain a condensing agent. As the condensing agent, for example, a carbodiimide condensing agent can be cited. As the carbodiimide condensing agent, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) can be cited. In addition to the condensing agent, the reaction solution may further contain a condensation aid. As the condensation aid, for example, 1-hydroxybenzotriazole (HOBT) can be cited.

[0041] (Post-processing process) As the post-treatment step, for example, a step of precipitating component A, a step of washing component A, etc. can be performed. As a method for precipitating component A, for example, a method of mixing an organic solvent such as ethanol in the reaction solution obtained after the reaction step can be cited. As a method for washing component A, a method of washing the precipitate containing component A with an ethanol aqueous solution or the like can be cited.

[0042] <Kyennepin> Genipin is a compound capable of forming a cross-linked structure by reacting with an amino group, and is a compound represented by the following formula. [Chemical formula 3]

[0043] (Mass ratio of genipin to component A) The ratio of the mass of genipin to the mass of component A (mass of genipin / mass of component A) may be 0.01 or more, 0.03 or more, or 0.05 or more. The ratio of the mass of genipin to the mass of component A may be 1.00 or less, 0.50 or less, 0.40 or less, 0.30 or less, or 0.25 or less.

[0044] (Form of Genipin) In the gel-forming material, genipin may be in liquid or solid form (e.g., powder form). The gel-forming material may include a solution containing genipin and a solvent or a powder containing genipin. As the solvent, water, a buffer solution, etc. may be listed. As the buffer solution, a phosphate buffer solution, etc. may be listed.

[0045] (Solution containing genipin) In the solution containing genipin, the content of genipin may be 0.1 mg or more, 1 mg or more, or 1.5 mg or more, and may be 25 mg or less, 15 mg or less, or 10 mg or less, based on 1 mL of the solvent.

[0046] (How to obtain Kinipin) Genipin can be used directly as a commercially available product.

[0047] <Medication> The gel-forming material may further contain a drug. The drug may be a water-soluble drug. Examples of the drug include peptides, proteins, and genes.

[0048] <Other ingredients> The gel-forming material may contain other components other than component A, genipin, and the drug. Examples of other components include inorganic salts such as water, ethanol, phosphate, sodium chloride, and potassium chloride.

[0049] <Method for gelling gel-forming material> The gel composition is formed by mixing the gel-forming material of the present invention, namely component A, and genipin. Examples of the gelation method include a method of mixing a solution containing component A and a solution containing genipin and heating to 30°C or higher, a method of mixing a powder containing genipin in a solution containing component A and heating to 30°C or higher, and a method of mixing a powder containing component A in a solution containing genipin and heating to 30°C or higher. From the viewpoint of suppressing the generation of lumps during gel formation, the gelation method is preferably a method of mixing a solution containing component A and a solution containing genipin.

[0050] (Gelation temperature conditions) The temperature (gelation temperature) at which the gel-forming material is gelled may be 30°C or higher or 35°C or higher, and may be 80°C or lower, 60°C or lower, 50°C or lower, 45°C or lower, or 40°C or lower.

[0051] <Method of administering the gel-forming material> As a method of administering the gel-forming material, administration by injection can be cited. When the gel-forming material is administered into the body by injection, a syringe having a syringe body and an injection needle can be used, wherein the syringe body can separately accommodate the first component containing at least component A and the second component containing at least genipin. When using this syringe, the first component and the second component are mixed before administration into the body (in the injection needle of the syringe) to form a gel composition at the location of administration.

[0052] <Biocompatible gel formation> Since the gel-forming material forms a gel composition by reaction between genipin, a cross-linking agent derived from natural plants (gardenia jasminoides) and known to have low toxicity, and component A, it is possible to form a biocompatible gel composition.

[0053] <Visualization of gel formation> Since the gel composition formed by the reaction of component A and genipin is colored, the formation of the gel composition can be confirmed visually, etc. Since the gel formed by the gel-forming material is visible, it is suitable for use in endoscopes, etc.

[0054] <In situ gel forming material 1> The in situ gel-forming material of one embodiment includes the above-mentioned component A. The in situ gel-forming material including the component A can form a gel in situ by reacting with the above-mentioned genipin.

[0055] <In situ gel forming material 2> The in situ gel-forming material of one embodiment includes the above-mentioned genipin. The in situ gel-forming material including genipin can form a gel in situ by reacting with the above-mentioned component A.

[0056] <Gel Composition> The gel composition of the present invention is formed by cross-linking the above-mentioned component A through genipin. That is, the gel composition contains component A and genipin, and a cross-linked structure is formed by the reaction of component A and genipin. The gel composition is formed by genipin, a cross-linking agent derived from nature (gardenia), which is known to have low toxicity, and component A, and therefore has biocompatibility. The gel composition is colored due to the formation of a cross-linked structure through the reaction of component A and genipin, and gelation is easily confirmed by visual observation.

[0057] (Gel composition form) The specific form of the gel composition can be applied to the specific form of the gel-forming material described above. The gel composition can contain the above-mentioned drugs and other components. The gel composition does not exclude the inclusion of impurities that are inevitably contained during the preparation.

[0058] <Method for producing gel composition> The gel composition can be obtained by a method including the step of gelling the above-mentioned gel-forming material. The gelling step can be performed, for example, by a method including mixing the above-mentioned component A with the above-mentioned genipin. The conditions for gelling can be as described above.

[0059] <Application of gel composition> Examples of uses of the gel composition include local sustained-release materials for drugs, anti-adhesion materials, hemostatic materials (e.g., hemostatic materials for endoscopic surgery), wound dressings, scaffold materials for cell culture, skin filling materials, and joint injection materials.

[0060] <Sustained-release preparations> The sustained-release preparation comprises: a first component comprising component A; a second component comprising genipin; and a drug. The drug may be contained in either or both of the first and second components. The sustained-release preparation contains the first and second components in a state before mixing. The above-mentioned other components may be contained in either or both of the first and second components. The sustained-release preparation contains the first and second components in a state before mixing. By mixing the first and second components immediately before or at the same time as administration to the body, a gel composition containing the drug is formed at a target site in the body. When the target site is in the body, the gel composition containing the drug is hydrolyzed in the body, and the drug is gradually released by deaggregation of the gel composition.

[0061] <Use of Component A and / or Genipin for Production of Gel-Forming Material> As one embodiment of the present invention, a use (application) of component A and genipin for the manufacture of a gel-forming material is provided. As one embodiment of the present invention, a use of component A for the manufacture of an in situ gel-forming material and a use of genipin for the manufacture of an in situ gel-forming material are provided. In these embodiments, the above-mentioned method of the gel-forming material can be applied without limitation.

[0062] <Component A and / or Genipin for Gel Formation> As one embodiment of the present invention, a component A for gel formation and genipin are provided. As one embodiment of the present invention, a component A for in situ gel formation and genipin for in situ gel formation are provided. In these embodiments, the above-mentioned gel-forming material can be applied without limitation. [Example]

[0063] Hereinafter, the present invention will be described in more detail based on Examples etc. However, the present invention is not limited to the following Examples.

[0064] [Production Example 1] <Method for producing modified hyaluronic acid having amino group (amino group modification rate 32%)> 1 g of hyaluronic acid (HyaluronsanHA-LQH manufactured by Kewpie Co., Ltd., with an average molecular weight of about 1.8 million) was dissolved in 100 mL of pure water to obtain a hyaluronic acid solution. 1530 mg of 1-hydroxybenzotriazole (HOBT), 1920 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and 13 g of adipic acid dihydrazide were added to the hyaluronic acid solution and reacted overnight. 300 mL of ethanol was slowly added to the obtained reaction solution to precipitate the modified hyaluronic acid, and washed 3 times with 100 mL of 80% ethanol to obtain an amino-containing modified hyaluronic acid (amino-containing modified HA) containing a modified group derived from adipic acid dihydrazide. The modification rate of the amino-containing modified HA is 32%.

[0065] [Production Example 2] <Method for producing modified hyaluronic acid having amino group (amino group modification rate 66%)> 0.5 g of hyaluronic acid (HyaluronsanHA-LQ manufactured by Kewpie Co., Ltd., with an average molecular weight of about 1.2 million) was dissolved in 50 mL of pure water to obtain a hyaluronic acid solution. 211 mg of 1-hydroxybenzotriazole (HOBT), 240 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and 166 mg of ethylenediamine dihydrochloride (EDA) were added to the hyaluronic acid solution and allowed to react overnight. 150 mL of ethanol was slowly added to the obtained reaction solution to precipitate the modified hyaluronic acid, and 50 mL of 80% ethanol was used to wash 3 times to obtain an amino-containing modified HA containing a modified group derived from ethylenediamine. The modification rate of the amino-containing modified HA is 66%.

[0066] [Production Examples 3 to 13] Except that the type and amount of hyaluronic acid (HA), the amounts of HOBT and EDC·HCl, and the type and amount of the raw material compound containing a modifying group were changed as shown in Table 1, an amino group-containing modified HA was obtained in the same manner as in Production Example 1 or 2. [Table 1]

[0067] <Method for measuring amino group modification rate> (Sample preparation) 7 mg of a sample and 1 mg of an internal standard substance, 4,4-dimethyl-4-silapentane sodium sulfonate (DSS), were dissolved in 0.7 mL of heavy water, and the mixture was transferred to an NMR sample tube and sealed.

[0068] (Measurement conditions) Apparatus: Varian NMR system 400NB (Varian Technologies Japan Ltd.) Observation frequency: 400MHz Temperature: 30℃ Standard: DSS (0ppm) Total times: 64 times

[0069] (Analysis method) right 1 The 2.0 ppm peak of CH3 derived from the N-acetyl group of the amino-containing modified hyaluronic acid, the 1.65 ppm peak of the modified group derived from adipic acid dihydrazide, and the 3.2 ppm peak of CH2 derived from the modified group from ethylenediamine in the H-NMR spectrum were integrated. The amino modification rate was calculated by the following formula based on the integrated values. Amino group modification rate (%) = (peak integral value of modified group of amino group-containing modified hyaluronic acid / 2) / (peak integral value of 2.0 ppm / 3) × 100

[0070] <Method for Determining Average Molecular Weight of Amino-Containing Modified Hyaluronic Acid> The obtained amino-containing modified hyaluronic acid was dissolved in a mobile phase at a concentration of 0.1%, and the average molecular weight was relatively determined by the following liquid chromatography analysis. A plurality of (purified) hyaluronic acids with known average molecular weights were used as standard substances, and a standard curve was prepared based on their retention times to calculate the average molecular weight of the amino-containing modified hyaluronic acid. Column: TSK guard column PWXL + TSK gel GMPW × 2 Column temperature: 40°C Measurement wavelength: 210nm Flow rate: 0.8 mL / min Sample injection volume: 20μL Analysis time: 40 minutes Mobile phase: 0.003 mol / L phosphate buffer-0.15 mol / L NaCl (pH 7.0) Photodiode array: 996 photodiode array manufactured by Waters Corporation, Japan HPLC system: manufactured by Waters Corporation, Japan, 2690 separation module

[0071] [Test example] <Method for producing gel composition> Using the amino-containing modified hyaluronic acid and genipin of manufacturing examples 1 to 13, gel compositions of test examples 1 to 18 were obtained. Specifically, the amino-containing modified hyaluronic acid and genipin in the amounts shown in Table 2 were dissolved in water or phosphate buffered saline (PBS), and the solutions were adjusted to various temperatures. The solution was turned upside down together with the container, and the time point when it was confirmed that the solution stayed in place for 1 minute without flowing was considered to have achieved gelation. "Room temperature" in Table 2 means 23°C.

[0072] [Table 2]

[0073] According to Test Examples 1 to 3, when the amino group-containing modified hyaluronic acid and genipin of Preparation Examples 1 and 2 were used, a gel composition could be obtained within 1 hour at 37° C., and was suitable for in situ use.

[0074] According to Test Example 4, even when the temperature during mixing is changed, it is found that a gel composition can be formed by using the component (A) and genipin.

[0075] By using component (A) and genipin when changing the amino modification rate of the amino group-containing modified hyaluronic acid, a gel can be formed in a short time at 37°C (Test Examples 5 to 9). By using component (A) and genipin when changing the average molecular weight, a gel can be formed in a short time at 37°C (Test Examples 10 to 13). By using component (A) and genipin when changing the type of amino group-containing modified HA, the amount of amino group-containing modified HA, or the amount of genipin, a gel can be formed in a short time at 37°C (Test Examples 14 to 18).

Claims

1. A gel-forming material, characterized in that: The invention comprises: at least one component A selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof; and Kinnipin.

2. The gel-forming material according to claim 1, characterized in that The amino group modification rate of the component A is 5% or more and 70% or less.

3. The gel-forming material according to claim 1 or 2, characterized in that The average molecular weight of the component A is 10,000 or more and 1,500,000 or less.

4. A material for in-situ gel formation, characterized in that: The present invention comprises at least one component selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof.

5. A material for in-situ gel formation, characterized in that: Contains Genipin.

6. A gel composition, characterized in that: The invention comprises at least one component A cross-linked by genipin, wherein the component A is selected from the group consisting of modified hyaluronic acid having an amino group and an average molecular weight of 10,000 or more and salts thereof.

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