Gel-forming materials and gel compositions

Abstract

The present invention relates to 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 a salt thereof, and a genipin.

Description

[Technical Field]

[0001] The present invention relates to gel-forming materials and gel compositions. [Background technology]

[0002] Materials that gel in situ after being implanted are attracting attention in the fields of drug carriers and tissue engineering because they can be used as medical materials that can be administered by injection into the body, enabling minimally invasive treatment. In addition to gelling relatively quickly in response to changes in temperature, pH, and chemical substances, biocompatibility and safety are also important considerations for materials that gel in situ. Among various gel-forming materials, hyaluronic acid exhibits excellent biocompatibility and safety for the human body. As a composition for obtaining hydrogels using hyaluronic acid, for example, Patent Document 1 discloses hyaluronic acid derivatives having a specific structure and their crosslinked forms, which can be used for the treatment of cytokine-related inflammation. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] Special Publication No. 2016-518464 [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] This invention provides a gel-forming material and gel composition that use hyaluronic acid and can gel in a short time at 37°C. Such gel-forming materials and gel compositions are suitable for medical materials that need to be administered into the body by injection and gel in situ.

[0005] One aspect of the present invention aims to provide a gel-forming material that can form a gel in a short time under conditions of 37°C. Another aspect of the present invention aims to provide a novel in situ gel-forming material. [Means for Solving the Problems]

[0006] The inventors have found that a gel-forming material comprising at least one component A selected from the group consisting of a modified hyaluronic acid having an amino group and having an average molecular weight of 10,000 or more and a salt thereof, and genipin, can solve the above problems, and have completed the present invention.

[0007] That is, the present invention relates to, for example, the following inventions. [1] A gel-forming material comprising at least one component A selected from the group consisting of a modified hyaluronic acid having an amino group and having an average molecular weight of 10,000 or more and a salt 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 selected from the group consisting of a modified hyaluronic acid having an amino group and having an average molecular weight of 10,000 or more and a salt thereof. [5] A material for in situ gel formation comprising genipin. [6] A gel composition in which at least one component A selected from the group consisting of a modified hyaluronic acid having an amino group and having an average molecular weight of 10,000 or more and a salt thereof is crosslinked with genipin. [[ID=3, ]] [Advantages of the Invention]

[0008] [[ID=4, ]]According to the present invention, it is possible to provide a gel-forming material capable of forming 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 material for in situ gel formation. [Modes for Carrying Out the Invention]

[0009] Hereinafter, embodiments for carrying out the present invention will be described in detail. However, the present invention is not limited to the following embodiments.

[0010] <Features of the Present Invention> (Material for gel formation) The present invention provides a material for gel formation comprising at least one component A selected from the group consisting of a modified hyaluronic acid having an amino group and having an average molecular weight of 10,000 or more and a salt thereof, and genipin. The material for gel formation of the present invention can be gelled within 1 hour under the condition of 37 ° C and can be suitably used for medical materials that need to be gelled in situ.

[0011] (Material for gel formation) The "material for gel formation" in this specification is a material containing one or more components that can form a gel composition by reaction with other components. The material for gel formation of the present invention may be one that forms a gel composition within 1 hour from the start of the reaction when reacted with other components at 37 ° C. The material for gel formation of the present invention comprises at least one component A selected from the group consisting of a modified hyaluronic acid into which an amino group has been introduced and a salt thereof, and genipin.

[0012] (Gel formation in situ) Since the material for gel formation of the present invention can form a gel within a short time of within 1 hour under the condition of 37 ° C, it is suitably used as a material for gel formation in situ. The "material for gel formation in situ" is a material for gel formation that can form a gel at the target location. The material for gel formation of the present invention can be sterilized in the state before gel formation (for example, in the state of a solution). Therefore, sterilization in the state after gelation is not necessarily required, and it is possible to administer the sterilized material for gel formation to the target location of the living body to form a gel.

[0013] (Component A) Component A is a compound or a salt thereof in which a functional group having an amino group (-NH2) is introduced into at least a part of hyaluronic acid, provided that those obtained by deacetylating the acetyl group of N-acetylglucosamine, which is a part of hyaluronic acid, to form an amino group are excluded. The gel-forming material may contain one or more of Component A.

[0014] (Structure 1 of Component A) Component A may be a compound or a salt thereof in which at least a part of the functional groups contained in some of the disaccharide units that make up the repeating unit of the hyaluronic acid are substituted with functional groups 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 hydroxy group at the C4 position and the hydroxy group at the C6 position of N-acetylglucosamine that constitutes hyaluronic acid, and the hydroxy group at the C2 position, the hydroxy group at the C3 position, and the carboxy group at the C6 position of glucuronic acid that constitutes 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

[0016] (Number and substitution position of the functional group having an amino group in formula (1)) In formula (1), at least one of R 1 ~R 5 is a functional group having an amino group. In formula (1), 1 to 4, 1 to 3, or 1 to 2 of R 1 ~R 5 may be functional groups having an amino group, and R 1 ~R 5Any one of them may be a functional group having an amino group. In formula (1), R 1 is a functional group having an amino group, and R 2 ~R 5 All of these can be hydrogen atoms.

[0017] (Structure of component A 3) Component A may be, for example, a compound represented by the following formula (1a) or a salt thereof. [ka] In formula (1a), R 1 and n are R in equation (1), respectively. 1 This is synonymous with n.

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

[0019] (Structure of R in equation (I) 1) R indicates a divalent group, X 1 It is a group that connects and NH2. The divalent group may contain 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 in the hydrocarbon group and alkylene group may be, for example, 1 or more, 2 or more, 10 or less, 8 or less, 6 or less, 4 or less, or 3 or less. R may contain a functional group other than a hydrocarbon group. The functional group other than a hydrocarbon group may be, for example, -CO-NH-.

[0020] (Structure of R in equation (I) 2) R is -R b -R a -R c -The base may be represented by R a R indicates an alkylene group, b and R cEach of these independently exhibits a single bond or *1-CO-NH-*2. Here, *1 is R a This shows the binding site, and *2 is X 1 Alternatively, it indicates a bonding site with an amino group. R may contain an alkylene group having 2 or more carbon atoms. When R contains an alkylene group having 2 or more carbon atoms, the resulting gel composition will have better elasticity. When R contains an alkylene group having 4 or fewer carbon atoms, the solubility in solvents such as water and PBS will be better.

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

[0022] (Specific examples of functional groups containing an amino group) Functional groups containing an amino group have a bond site that is R 1 In this case, 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. Functional groups having an amino group have a bond site R 2 ~R 5 In this case, it may be -O-CH2-CH2-NH2, -(C=O)-CH2-NH2, or -CH2-CH2-NH2.

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

[0024] (Suitable upper limit for the 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, its solubility in solvents such as water or PBS becomes even better.

[0025] (A suitable lower limit for the average molecular weight of component A) The average molecular weight of component A is 10,000 or more, and may be 20,000 or more, 30,000 or more, 40,000 or more, 50,000 or more, 60,000 or more, 70,000 or more, 80,000 or more, 90,000 or more, or 100,000 or more. When the average molecular weight of component A is 10,000 or more, the elasticity of the resulting gel composition will be even better.

[0026] (Preferred numerical range for the 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 measuring average molecular weight) The average molecular weight can be measured by the following method. First, using a gel filtration column, several (purified) hyaluronic acid samples (reference substances) with known molecular weights are analyzed by liquid chromatography, and a calibration curve is created from their retention times. Similarly, the molecular weight of component A, the target of measurement, is determined by liquid chromatography analysis, and its molecular weight is calculated using the prepared calibration curve.

[0028] (Measurement device and measurement conditions for average molecular weight) A Waters Japan 2690 separation module is used as the liquid chromatography analyzer. A Waters Japan 996 photodiode array is used as the photodiode array. One TSK Guard column PWXL (Tosoh Corporation) and two TSK Gel GMPW (Tosoh Corporation) are connected in series in the order listed. For the measurement of average molecular weight, the following conditions are used: column temperature 40°C, measurement wavelength 210 nm, flow rate 0.8 mL / min, sample injection volume 20 μL, analysis time 40 minutes, and mobile phase 0.003 mol / L phosphate buffer - 0.15 mol / L NaCl (pH 7.0). Details of other test conditions may be as described in the examples below.

[0029] (Definition of amino group modification rate) The amino group modification rate of component A refers to the number of amino groups contained in one unit, with each disaccharide unit constituting hyaluronic acid being considered as one unit. Specifically, it refers to the ratio (%) of the number of amino groups contained in one unit to one unit, with the unit being considered as 100%. In this specification, "disaccharide unit constituting hyaluronic acid" refers to one unit composed of adjacently bonded disaccharides (glucuronic acid and N-acetylglucosamine) that constitute hyaluronic acid.

[0030] (Method for measuring the amino group modification rate) The amino group modification rate is, 1 Measurement can be performed using 1H-NMR. Specific examples of sample preparation and measurement methods are described in the examples below.

[0031] (Lower limit of amino group modification rate) The amino group 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 group modification rate is 10% or more, the gel-forming ability becomes even better even at low concentrations of component A in the gel-forming material, such as less than 1%. The amino group 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, as this enables gel formation in an even shorter time.

[0032] (Suitable upper limit for amino group modification rate) The amino group modification rate may be 90% or less, 85% or less, 80% or less, 75% or less, or 70% or less. When the amino group modification rate is 70% or less, the solubility in solvents such as water and PBS becomes even better.

[0033] (Preferred numerical range for amino group modification rate) From the viewpoint of achieving even better solubility in solvents such as water and PBS, the amino group modification rate is preferably 5% to 70%, and more preferably 10% to 60%.

[0034] (Form of component A) In gel-forming materials, component A may be liquid or solid (e.g., powder). The gel-forming material may contain a solution containing component A and a solvent, or a powder containing component A. Examples of solvents include water and buffer solutions. Examples of buffer solutions include phosphate buffer solutions.

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

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

[0037] (Raw material compound) The starting compound is given by formula (Ia):X 2 The compound may be represented by -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 This indicates a group that can react with functional groups in hyaluronic acid. Examples of groups that can react with functional groups in hyaluronic acid include amino groups (-NH2). The amount of raw material compound used is appropriately adjusted according to the amino group modification rate of the target component A.

[0038] (Specific examples of raw material compounds) Examples of raw material compounds include ethylenediamine, adipic acid dihydrazide, methanediamine, 1,3-propanediamine (1,3-diaminopropane), 1,4-butanediamine, 2-aminoethanol, glycine, and 2-chloroethyldiamine.

[0039] (Solvent in the reaction process) The reaction solution may contain a solvent. Examples of solvents include water, or a mixture 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. Examples of condensing agents include carbodiimide-based condensing agents. An example of a carbodiimide-based condensing agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl). In addition to the condensing agent, the reaction solution may further contain a condensation auxiliary agent. An example of a condensation auxiliary agent is 1-hydroxybenzotriazole (HOBT).

[0041] (Post-processing steps) Post-processing steps may include, for example, a step to precipitate component A, a step to wash component A, etc. A method for precipitating component A is to mix an organic solvent such as ethanol with the reaction solution obtained after the reaction step. A method for washing component A is to wash the precipitate containing component A with an aqueous solution of ethanol or the like.

[0042] <Genipin> Genipine is a compound that can form a cross-linked structure through reaction with an amino group, and is represented by the following formula. [ka]

[0043] (Mass ratio of genipin to component A) The ratio of the mass of genipine to the mass of component A (mass of genipine / mass of component A) may be 0.01 or greater, 0.03 or greater, or 0.05 or greater. The ratio of the mass of genipine 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] (Genipin morphology) In gel-forming materials, genipin may be in liquid or solid form (e.g., powder). The gel-forming material may contain a solution containing genipin and a solvent, or a powder containing genipin. Examples of solvents include water and buffer solutions. Examples of buffer solutions include phosphate buffer solutions.

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

[0046] (How to obtain Genipin) Genipin can be used as is, as it is obtained from the market.

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

[0048] <Other ingredients> The gel-forming material may contain component A, genipin, and other components that are not considered drugs. Examples of other components include water, ethanol, phosphates, sodium chloride, potassium chloride, and other inorganic salts.

[0049] <Method for gelling materials> A gel composition is formed by mixing component A and genipin, which are gel-forming materials of the present invention. Methods for gelation include mixing a solution containing component A with a solution containing genipin and heating to 30°C or higher, mixing a powder containing genipin with a solution containing component A and heating to 30°C or higher, and mixing a powder containing component A with a solution containing genipin and heating to 30°C or higher. From the viewpoint of suppressing the formation of clumps during gel formation, the method of gelation is preferably the method of mixing a solution containing component A with a solution containing genipin.

[0050] (Temperature conditions for gelation) The temperature at which the gel-forming material is gelled (gelling temperature) 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 dispensing gel-forming material> As a method of administering the gel-forming material, administration by injection can be mentioned. When the gel-forming material is administered into a living body by injection, it can be carried out using a syringe comprising a syringe body capable of separately accommodating a first component containing at least component A and a second component containing at least genipin, and an injection needle. When using this syringe, the first component and the second component are mixed immediately before administration into the living body (inside the injection needle of the syringe), and a gel composition is formed at the administration site.

[0052] <Gel formation with biocompatibility> The gel-forming material is a crosslinking agent derived from natural (gardenia), and since it forms a gel composition by the reaction of genipin, which is known to have low toxicity, with component A, it is possible to form a gel composition having biocompatibility.

[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 or the like. Since the gel formation by the gel-forming material is visualized, it is useful in applications such as endoscopes.

[0054] <In-situ gel-forming material 1> The in-situ gel-forming material of one embodiment contains component A described above. The in-situ gel-forming material containing component A can form a gel in-situ by reaction with the above-described genipin.

[0055] <In-situ gel-forming material 2> The in-situ gel-forming material of one embodiment contains the above-described genipin. The in-situ gel-forming material containing genipin can form a gel in-situ by reaction with the above-described component A.

[0056] <Gel composition> The gel composition of the present invention is formed by crosslinking component A with genipine. That is, the gel composition contains component A and genipine, and a crosslinked structure is formed by the reaction between component A and genipine. The gel composition is biocompatible because it is formed from genipine, a crosslinking agent derived from natural materials (gardenia) and known to have low toxicity, and component A. The gel composition is colored because a crosslinked structure is formed by the reaction between component A and genipine, making it easy to visually confirm gelation.

[0057] (Modes of gel composition) Specific embodiments of the gel composition can be those of the specific embodiments of the gel-forming material described above. The gel composition may contain the drugs and other components described above. The gel composition is not excluded from containing impurities that are inevitably present during preparation.

[0058] <Method for producing gel composition> The gel composition can be obtained by a method that includes a step of gelling the gel-forming material described above. The gelling step can be carried out, for example, by a method that includes mixing component A described above and genipin described above. The conditions for gelling may be as described above.

[0059] <Uses of gel compositions> Applications of the gel composition include sustained-release agents for local drug delivery, adhesion inhibitors, hemostatic agents (e.g., hemostatic agents for endoscopic surgery), wound dressings, scaffolds for cell culture, skin fillers, and joint injection materials.

[0060] <Sustained-release formulation> The sustained-release formulation contains a first component containing component A, a second component containing genipine, and a drug. The drug may be contained in either or both of the first and second components. The sustained-release formulation contains the first and second components in their unmixed state. Other components mentioned above may be contained in either or both of the first and second components. The sustained-release formulation contains the first and second components in their unmixed state. By mixing the first and second components immediately before or during administration to a living organism, a gel composition containing the drug is formed at the target site in the living organism. If the target site is within the living organism, the gel composition containing the drug undergoes hydrolysis or other processes within the organism, and the drug is gradually released as the gel composition disintegrates.

[0061] <Use of component A and / or genipin for the manufacture of gel-forming materials> One embodiment of the present invention provides the use (application) of component A and genipine for the manufacture of a gel-forming material. Another embodiment of the present invention provides the use of component A for the manufacture of a gel-forming material in situ, and the use of genipine for the manufacture of a gel-forming material in situ. In these embodiments, the embodiments of the gel-forming material described above can be applied without limitation.

[0062] <Component A and / or genipin used for gel formation> One embodiment of the present invention provides component A and genipin for use in gel formation. Another embodiment of the present invention provides component A and genipin for use in situ gel formation. In these embodiments, the embodiments of the gel-forming material described above can be applied without limitation. [Examples]

[0063] The present invention will be described more specifically below based on examples. However, the present invention is not limited to the following examples.

[0064] [Manufacturing Example 1] <Method for producing modified hyaluronic acid with amino groups (amino group modification rate 32%)> 1 g of hyaluronic acid (Hyaluronic Acid HA-LQH, manufactured by Kewpie Corporation, average molecular weight approximately 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 resulting reaction solution to precipitate the modified hyaluronic acid, which was then washed three times with 100 mL of 80% ethanol to obtain amino group-containing modified hyaluronic acid (amino group-containing modified HA) containing a modifying group derived from adipic acid dihydrazide. The modification rate of the amino group-containing modified HA was 32%.

[0065] [Manufacturing Example 2] <Method for producing modified hyaluronic acid with amino groups (amino group modification rate 66%)> 0.5 g of hyaluronic acid (Hyaluronic Acid HA-LQ, manufactured by Kewpie Corporation, average molecular weight approximately 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 reacted overnight. 150 mL of ethanol was slowly added to the resulting reaction solution to precipitate the modified hyaluronic acid, which was then washed three times with 50 mL of 80% ethanol to obtain amino group-containing modified HA containing a modifying group derived from ethylenediamine. The modification rate of this amino group-containing modified HA was 66%.

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

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

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

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

[0070] <Method for measuring the average molecular weight of amino group-modified hyaluronic acid> The obtained amino group-containing modified hyaluronic acid was dissolved in a mobile phase at a concentration of 0.1%, and its relative average molecular weight was measured by the following liquid chromatography analysis. As reference substances, several (purified) hyaluronic acid samples with known average molecular weights were used, and a calibration curve was created from their retention times to calculate the average molecular weight of the amino group-containing modified hyaluronic acid. Column: TSK Guard Column PWXL + TSK Gel GMPW x 2 Column temperature: 40℃ Measurement wavelength: 210nm Flow rate: 0.8mL / min Sample injection volume: 20 μL 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 Japan Co., Ltd. HPLC system: Waters Japan Co., Ltd., 2690 separation module

[0071] [Example Test] <Method for producing gel composition> Using the amino group-containing modified hyaluronic acid and genipin from Production Examples 1-13, gel compositions from Test Examples 1-18 were obtained. Specifically, the amounts of amino group-containing modified hyaluronic acid and genipin shown in Table 2 were dissolved in water or phosphate-buffered saline (PBS) to prepare solutions adjusted to each temperature. The solution was turned upside down in the container and left for 1 minute. Gelation was defined as the point at which the solution remained in place and did not flow. In Table 2, "room temperature" refers to 23°C.

[0072] [Table 2]

[0073] From Test Examples 1-3, it can be seen that using the amino group-containing modified hyaluronic acid and genipin from Production Examples 1 and 2, a gel composition can be obtained within 1 hour at 37°C, making it suitable for in situ use.

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

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

Claims

1. At least one component A selected from the group consisting of modified hyaluronic acid having an amino group and salts thereof, Genipin, and, The ratio of the mass of genipin to the mass of component A is 0.05 or more and 0.25 or less. The amino group modification rate of component A is 20% or more and 70% or less. A gel-forming material wherein the average molecular weight of component A is 200,000 or more and 1,000,000 or less.

2. The gel-forming material according to claim 1, which is a medical material.

3. The gel-forming material according to claim 1 or 2, which is an adhesion prevention material or a hemostatic material.

4. It contains at least one component A selected from the group consisting of modified hyaluronic acid having an amino group and salts thereof, This material forms a gel in situ upon reaction with genipin. The ratio of the mass of genipin to the mass of component A is 0.05 or more and 0.25 or less. The amino group modification rate of component A is 20% or more and 70% or less. An in-situ gel-forming material wherein the average molecular weight of component A is 200,000 or more and 1,000,000 or less.

5. A medical material, the in-situ gel-forming material according to claim 4.

6. An in-situ gel-forming material according to claim 4 or 5, which is an adhesion prevention material or a hemostatic material.

7. Contains genipin, This material forms a gel in situ by reacting with at least one component A selected from the group consisting of modified hyaluronic acid having an amino group and salts thereof. The ratio of the mass of genipin to the mass of component A is 0.05 or more and 0.25 or less. The amino group modification rate of component A is 20% or more and 70% or less. An in-situ gel-forming material wherein the average molecular weight of component A is 200,000 or more and 1,000,000 or less.

8. The in-situ gel-forming material according to claim 7, which is a medical material.

9. An in-situ gel-forming material according to claim 7 or 8, which is an adhesion prevention material or a hemostatic material.

10. At least one component A, selected from the group consisting of modified hyaluronic acid having an amino group and its salts, is crosslinked with genipin. The ratio of the mass of genipin to the mass of component A is 0.05 or more and 0.25 or less. The amino group modification rate of component A is 20% or more and 70% or less. A gel composition in which the average molecular weight of component A is 200,000 or more and 1,000,000 or less.

11. The gel composition according to claim 10, which is a medical material.

12. The gel composition according to claim 10 or 11, which is an adhesion prevention material or a hemostatic material.

Images