Adhesive, wound dressing material, Anti-adhesion material, hemostatic agent, sealant, and filler

Abstract

To provide an adhesive that exhibits superior tissue adhesiveness in aqueous environments, along with excellent biocompatibility.SOLUTION: An adhesive comprises a first agent including a gelatin derivative having a structure represented by formula (1), and a second agent including a hydroxyl group-containing compound. In formula (1), Gltn represents a gelatin residue, X represents one selected from the group consisting of -C(O)-NH-, -NH-C(O)-, and -NH-, and L represents a single bond or a divalent linking group.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to adhesives, wound dressings, adhesion inhibitors, hemostats, sealants, and fillers. [Background technology]

[0002] Tissue adhesives, surgical sealants, etc. using gelatin derivatives obtained by introducing hydrophobic groups into gelatin are known. For example, Patent Document 1 reports a surgical sealant consisting of a first agent containing hydrophobized gelatin into which a linear alkyl group has been introduced, and a second agent containing a hydroxyl group-containing compound. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2016 / 117569 Summary of the Invention [Problem to be solved by the invention]

[0004] It has been revealed that the above-mentioned surgical sealant has biocompatibility and exhibits excellent adhesion to biological tissues such as the lungs, large intestine, etc. However, further improvement is required when the sealant is used on biological tissues in an aqueous environment where a large amount of blood or exudates is present.

[0005] The present invention is devised to solve the above problems, and provides an adhesive that has both high biocompatibility and adhesive strength and can be used on tissues such as the large intestine in an aqueous environment where bleeding and exudate occur. [Means for solving the problem]

[0006] As a result of extensive investigations aimed at achieving the above object, the present inventors have found that the above object can be achieved by the following configuration.

[0007] [1] An adhesive comprising a first agent containing a gelatin derivative having a structure represented by formula (1) described below, and a second agent containing a hydroxyl group-containing compound. [2] The adhesive according to [1], wherein in formula (1), the divalent linking group includes at least one selected from the group consisting of an alkylene group having 1 to 10 carbon atoms, a phenylene group, and a polyethylene oxide having 1 to 10 repeating numbers. [3] The adhesive according to [1] or [2], wherein the hydroxyl group-containing compound is a compound having at least two hydroxyl groups. [4] The adhesive according to any one of [1] to [3], wherein the structure of the hydroxyl group-containing compound is a structure represented by formula (2) described below. [5] The adhesive according to [4], wherein in formula (2), the divalent linking group is at least one selected from the group consisting of an alkylene group having 1 to 10 carbon atoms and a polyethylene oxide having 1 to 10 repeating units. [6] The adhesive according to [4] or [5], wherein in formula (2), the hydroxyl group in the benzene ring is at the meta position and / or para position relative to the bonding position of L1. [7] The adhesive according to any one of [1] to [3], wherein the hydroxyl group-containing compound is at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, polysaccharides, polyhydric alcohols, sugar alcohols, polyphenols and polyvinyl alcohols. [8] A wound dressing comprising the adhesive according to any one of [1] to [7]. [9] An anti-adhesion material comprising the adhesive described in any one of [1] to [7].

[10] A hemostatic material comprising the adhesive described in any one of [1] to [7].

[11] A sealant comprising the adhesive according to any one of [1] to [7].

[12] A filler comprising the adhesive according to any one of [1] to [7]. Effect of the Invention

[0008] The adhesive of the present invention has both high biocompatibility and adhesive strength, and can be used on biological tissue in an environment where moisture such as blood or exudate is present, for example, after colon surgery. [Brief description of the drawings]

[0009] [Figure 1] 4 is a conceptual diagram showing the curing reaction of the adhesive of the present embodiment. FIG. [Diagram 2] FIG. 2 is a diagram illustrating a method for synthesizing a gelatin derivative in an example. [Diagram 3] FIG. 2 is a diagram illustrating a method for synthesizing a hydroxyl group-containing compound in an example. [Figure 4] FIG. 2 is a diagram illustrating a pressure resistance test method in the examples. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The present invention will be described in detail below. The following description of the components may be based on a representative embodiment of the present invention, but the present invention is not limited to such an embodiment. In this specification, a numerical range expressed using "to" means a range that includes the numerical values ​​before and after "to" as the lower and upper limits.

[0011] [glue] The adhesive of this embodiment is a two-part adhesive having a first part containing a gelatin derivative and a second part containing a hydroxyl group-containing compound. The first part and the second part are packaged separately and provided. When used, the first part and the second part are mixed to form the skeleton of the hardened material (gel-like hardened adhesive material). The hardening reaction is typically a reaction between the boronic acid group of the gelatin derivative and the hydroxyl group of the second part.

[0012] 1. First agent The first agent of the present embodiment contains a gelatin derivative containing a structure represented by the following formula (1), and may further contain a solvent.

[0013] <Gelatin derivatives> The gelatin derivative having the structure represented by formula (1) has at least one boronic acid group (-B(OH)2) bonded to the raw material gelatin (raw material gelatin) via -XL-.

[0014] [ka] In formula (1), X represents one selected from the group consisting of -C(O)-NH-, -NH-C(O)-, and -NH-, and L represents a single bond or a divalent linking group.

[0015] The adhesive of this embodiment is a derivative of gelatin, a biopolymer, and therefore has high biocompatibility. Because the gelatin derivative contains a boronic acid group, when mixed with the hydroxyl group of the second agent, a dynamic covalent bond is formed between the boronic acid and the hydroxyl group (see FIG. 1), and the adhesive exhibits strong adhesive strength to biological tissue and also has high pressure resistance.

[0016] In formula (1), it is preferable that the divalent linking group contains at least one selected from the group consisting of an alkylene group having 1 to 10 carbon atoms, a phenylene group (a group in which two hydrogen atoms have been removed from a benzene ring, -(C6H4)-), and a polyethylene oxide having a repeating number of 1 to 10. Among these, from the viewpoint of stabilizing the boronic acid, it is preferable that the divalent linking group contains a phenylene group (for example, a 1,4-phenylene group in which two hydrogen atoms at the para position have been removed) directly bonded to the boron atom (B).

[0017] In formula (1), the nitrogen atom (N) directly bonded to the gelatin residue Gltn is derived from the ε-amino group of lysine (Lys) in gelatin. The NH structure in formula (1) is, for example, at 3300 cm in the FT-IR (Fourier transform infrared spectrometer) spectrum. -1 It can be detected by nearby bands.

[0018] The gelatin residue Gltn is a structure derived from the raw gelatin. Details of the raw gelatin will be explained in the method for producing gelatin derivatives described below.

[0019] Here, the molar ratio of the content of imino groups (-NH-) or amide groups (-NHC(O)-) to which boronic acid groups are bonded (introduced) in the gelatin derivative to the content of amino groups (primary amino groups, -NH2) in the raw material gelatin is defined as the "boronic acid group introduction rate."

[0020] The introduction rate of boronic acid groups in the gelatin derivative is not particularly limited, but from the viewpoint of obtaining an adhesive having a more excellent effect of the present embodiment, it is preferably, for example, 30 mol% to 90 mol%, or 40 mol% to 80 mol%. In other words, the imino group (or amide group) / amino group (molar ratio) in the gelatin derivative is preferably 30 / 70 to 90 / 10, or 40 / 60 to 80 / 20.

[0021] In the embodiment, the introduction rate of boronic acid groups is calculated from the values ​​obtained by quantifying the number of amino groups in raw gelatin and the number of amino groups in a gelatin derivative by the 2,4,6-trinitrobenzenesulfonic acid method (TNBS method) using the following formula. Boronic acid group introduction rate of gelatin derivative (mol%) = [Number of amino groups in raw gelatin - Number of amino groups in gelatin derivative] / [Number of amino groups in raw gelatin] x 100

[0022] The number of boronic acid groups introduced into one molecule of gelatin derivative is not particularly limited, and may be appropriately adjusted so that the boronic acid introduction rate is a predetermined value based on the molecular weight of raw material gelatin. For example, the number of boronic acid groups in one molecule of gelatin derivative may be 5 to 30, 7 to 20, or 10 to 15.

[0023] The molecular weight of the gelatin derivative is not particularly limited, and is determined by the molecular weight of the raw gelatin and the type and amount (number) of the introduced boronic acid group. Therefore, the range of the weight average molecular weight (Mw) of the gelatin derivative is almost the same as the range of the weight average molecular weight (Mw) of the raw gelatin described later.

[0024] The content of the gelatin derivative in the first agent is not particularly limited, but the concentration of the gelatin derivative in the first agent (gelatin derivative / solvent, rounded to the nearest four decimal places) is preferably 0.010 to 0.300 g / mL (rounded to the nearest four decimal places), more preferably exceeds 0.050 g / mL, even more preferably is 0.075 g / mL or more, particularly preferably exceeds 0.075 g / mL, preferably is 0.150 g / mL or less, more preferably is less than 0.150 g / mL, and particularly preferably is 0.100 g / mL or less.

[0025] The gelatin derivative may be composed of only one type of gelatin derivative, or may be a mixture of two or more types of gelatin derivatives. When the first agent contains two or more types of gelatin derivatives, the total content is preferably within the above numerical range.

[0026] <Method of producing gelatin derivatives> The method for producing the gelatin derivative is not particularly limited, and any known method can be used.

[0027] For example, a method of reacting an ε-amino group of gelatin with an aldehyde or ketone, binding a phenylboronic acid group via a Schiff base, and reducing the Schiff base to obtain a gelatin derivative is exemplified. This method is described, for example, in paragraphs 0029 to 0031 of JP 2019-216755 A. According to this method, a gelatin derivative in which a phenylboronic acid group is bound to a gelatin residue via an imino group is obtained. This boronic acid group is derived from the aldehyde or ketone used in the reaction.

[0028] Another method is to react an acid halide or a chloroformate compound with an ε-amino group of gelatin in the presence of a base such as triethylamine to obtain an amide. This method is described, for example, in paragraphs 0072 to 0080 of WO 2014 / 112208. According to this method, a gelatin derivative is obtained in which a boronic acid group is bound to a gelatin residue via an amide bond (including an imino group). This boronic acid group is derived from the acid halide or chloroformate compound used in the reaction.

[0029] When a large excess of a poor solvent, for example cold ethanol, is added to the reaction solution obtained above, the gelatin derivative precipitates, which is then filtered off and dried to obtain a powdered gelatin derivative. The gelatin derivative may be washed with ethanol or the like before drying.

[0030] The raw material gelatin used in the production of gelatin derivatives (hereinafter also referred to as "Org gelatin") is typically gelatin that does not have a boronic acid group introduced therein (is not derivatized).

[0031] The molecular weight of Org gelatin is not particularly limited, but the weight average molecular weight is preferably from 10,000 to 300,000, more preferably from 20,000 to 200,000, and particularly preferably from 30,000 to 100,000.

[0032] Org gelatin can be used without any particular limitation, regardless of whether it is naturally derived, chemically synthesized, fermented, or obtained by genetic recombination. Among them, gelatin prepared from naturally derived collagen is preferred. Examples of naturally derived collagen include those derived from mammals such as cows and pigs, and those derived from fish such as sea bream, sturgeon, salmon, and cod.

[0033] When the adhesive is used as a liquid, it preferably has excellent fluidity at the temperature of use (eg, living body temperature) from the standpoint of ease of handling. In this respect, the Org gelatin is preferably fish-derived gelatin, and in particular, gelatin derived from cold-water fish such as salmon and Alaska pollock. In addition, "using as a liquid" means that either the first agent or the second agent, or both, are liquids containing a solvent, or that both the first agent and the second agent are solids that are mixed with a solvent at the time of use.

[0034] Fish-derived gelatin, particularly cold-water fish gelatin, preferably has, per 1000 amino acids as constituent units, the number of units derived from hydroxyproline of 80 or less and / or the number of units derived from proline of 110 or less. Gelatin satisfying these conditions has superior fluidity at room temperature, and therefore, when used in the first agent, an adhesive with excellent handleability can be obtained.

[0035] The Org gelatin may be either acid-processed gelatin or alkali-processed gelatin. The first agent may contain two or more different types of gelatin as the Org gelatin. The two or more different types mean that one or more of the origin, molecular weight, processing method, etc. are different.

[0036] <Solvent> The first agent may further contain a solvent. Examples of the solvent include aqueous solvents, and examples of the aqueous solvent that can be used include ultrapure water, physiological saline, various inorganic salt buffers such as boric acid, phosphoric acid, and carbonate, or mixtures thereof. The aqueous solvent is preferably a phosphate buffer solution having a pH of 6 to 8, and more preferably a carbonate buffer solution having a pH of 9 to 10. The aqueous solvent is preferably used in an amount such that the solid content of the first agent is 0.050 to 0.300 g / mL.

[0037] <Additives> The first agent may be composed of only the gelatin derivative, or may be composed of only the gelatin derivative and the solvent, or may contain other additives within the range in which the effects of the present embodiment are achieved. Examples of additives include colorants, pH adjusters, and preservatives. For example, a colorant (e.g., brilliant blue) may be added to make it easier to see where the adhesive is applied. The amount of the additive may be, for example, 10 to 100 μg / mL.

[0038] In addition, since the adhesive of this embodiment (particularly the first agent) has a low viscosity, it is preferably used in the form of a mist sprayed from a sprayer and applied; however, by adding a viscosity adjuster, the viscosity can be easily increased and dripping of the coating film before hardening can be suppressed, so that it has the advantage of being able to adjust the viscosity according to the application area and use.

[0039] The adhesive contains a gelatin derivative having a boronic acid group introduced therein (a gelatin derivative containing a structure represented by the above formula (1)), but so long as it contains the gelatin derivative, it may also contain "other" gelatin and / or "other" gelatin derivatives. That is, when the first agent contains a derivative of Alaska pollock gelatin, the first agent may contain, for example, Alaska pollock gelatin, porcine gelatin, and / or porcine gelatin derivatives.

[0040] 2. Second agent The second agent contains a hydroxyl group-containing compound and may also contain a solvent.

[0041] <Hydroxyl group-containing compounds> The hydroxyl group-containing compound is typically a compound having at least two hydroxyl groups in one molecule that can react with the boronic acid group of the gelatin derivative.

[0042] The structure of the hydroxyl group-containing compound is not particularly limited as long as it is a compound having at least two hydroxyl groups in one molecule, but it is preferable that the structure has a structure represented by formula (2). The hydroxyl group-containing compound has a benzene ring having at least one hydroxyl group (hereinafter, appropriately referred to as a "specific phenol group") bonded to the raw material gelatin (raw material gelatin) via -Y-L1-.

[0043] [ka] In formula (2), Gltn represents a gelatin residue, Y represents one selected from the group consisting of -C(O)-NH-, -NH-C(O)-, and -NH-, L1 represents a single bond or a divalent linking group, and n is 1 to 5.

[0044] In the specific phenol group of formula (2), n is 1 to 5, preferably 1 to 3, and more preferably n = 2. The position of the hydroxyl group is not particularly limited, but when n = 1 to 3, the hydroxyl group in the benzene ring is preferably at the meta position and / or para position relative to the bonding position of L1.

[0045] More preferred embodiments of the specific phenol group include those represented by the following formula: In the following formula, * indicates the linking position with L1 in formula (2). Among these, the specific phenol group is preferably a catechol group having two hydroxyl groups, in that an adhesive having better effects of the present embodiment can be obtained.

[0046] [ka]

[0047] The divalent linking group of L1 in formula (2) includes, for example, -C(O)-, -C(O)O-, -OC(O)-, -O-, -S-, -N(R)- (R represents a hydrogen atom or a monovalent organic group (preferably a hydrocarbon group having 1 to 20 carbon atoms)), an alkylene group (preferably an alkylene group having 1 to 10 carbon atoms), an alkenylene group (preferably an alkenylene group having 2 to 10 carbon atoms), a polyethylene oxide having a repeating number of 1 to 10 (-(CH2CHO) n Among these, -C(O)-, an alkylene group having 1 to 10 carbon atoms, polyethylene oxide having a repeating number of 1 to 10, and combinations thereof are preferred.

[0048] For example, in formula (2), L1 may be an alkylene group having 1 to 10 carbon atoms, preferably an alkylene group having 1 to 3 carbon atoms, more preferably a methylene group. In this case, the hydroxyl group-containing compound of this embodiment has a structure in which a specific phenol group is bonded to gelatin (raw material gelatin) via an imino group (-NH-) and an alkylene group. L1 may also be a polyethylene oxide having a repeat number of 1 to 10. In this case, the hydroxyl group-containing compound of this embodiment has a structure in which a specific phenol group is bonded to gelatin (raw material gelatin) via an imino group (-NH-) and a polyethylene oxide.

[0049] In formula (2), the nitrogen atom (N) directly bonded to the gelatin residue Gltn is derived from the ε-amino group of lysine (Lys) in gelatin. The NH structure in formula (2) can be detected by the same method as that for the gelatin derivative in the first agent described above. The gelatin residue Gltn is a structure derived from the raw gelatin, and the raw gelatin can be the same as that used in the first agent.

[0050] Here, the molar ratio of the content of imino groups (-NH-) or amide groups (-NHC(O)-) to which specific phenolic groups (e.g., catechol groups) are bonded (introduced) in the gelatin derivative to the content of amino groups (primary amino groups, -NH2) in the raw material gelatin is defined as the "specific phenolic group introduction rate."

[0051] The specific phenol group introduction rate is not particularly limited, but from the viewpoint of obtaining an adhesive having a more excellent effect of this embodiment, it is preferably, for example, 30 mol% to 90 mol%, or 40 mol% to 80 mol%. In other words, the imino group (or amide group) / amino group (molar ratio) in the gelatin derivative is preferably 30 / 70 to 90 / 10, or 40 / 60 to 80 / 20.

[0052] In the embodiment, the specific phenol group introduction rate is calculated from the values ​​obtained by quantifying the number of amino groups in the raw gelatin and the number of amino groups in the gelatin derivative (hydroxyl group-containing compound) by the 2,4,6-trinitrobenzenesulfonic acid method (TNBS method) using the following formula. Specific phenol group introduction rate (mol%) of gelatin derivative (hydroxyl group-containing compound) = [Number of amino groups in raw gelatin - Number of amino groups in gelatin derivative] / [Number of amino groups in raw gelatin] x 100

[0053] The number of specific phenol groups (e.g., catechol groups) introduced into one molecule of the hydroxyl group-containing compound represented by formula (2) is not particularly limited, and may be appropriately adjusted so that the specific phenol group introduction rate is a predetermined value based on the molecular weight of the raw material gelatin. For example, the number of specific phenol groups in one molecule of the hydroxyl group-containing compound may be 5 to 30, 7 to 20, or 10 to 15.

[0054] The molecular weight of the hydroxyl group-containing compound represented by formula (2) is not particularly limited, and is determined by the molecular weight of the raw material gelatin and the type and amount (number) of the introduced hydrophobic group (specific phenol group, etc.). Therefore, the possible range of the weight average molecular weight (Mw) of the hydroxyl group-containing compound represented by formula (2) is almost the same as the possible range of the weight average molecular weight (Mw) of the raw material gelatin.

[0055] The content of the hydroxyl-containing compound represented by formula (2) in the second agent is not particularly limited, but the concentration of the hydroxyl-containing compound in the second agent (hydroxyl-containing compound / solvent, rounded to the nearest three decimal places) is preferably 0.010 to 0.300 g / mL (rounded to the nearest three decimal places), more preferably greater than 0.050 g / mL, even more preferably 0.075 g / mL or more, particularly preferably greater than 0.075 g / mL, preferably 0.150 g / mL or less, more preferably less than 0.150 g / mL, and particularly preferably 0.100 g / mL or less.

[0056] The method for producing the hydroxyl group-containing compound represented by formula (2) (gelatin derivative having specific phenol group) is not particularly limited, and a known method can be used. For example, in the method for producing the gelatin derivative contained in the first agent described above, the specific phenol group can be introduced into the ε-amino group of the raw material gelatin instead of the boronic acid group (typically, phenylboronic acid group).

[0057] The hydroxyl group-containing compound of this embodiment is not limited to the hydroxyl group-containing compound represented by formula (2) described above (gelatin derivative having a specific phenol group). For example, the hydroxyl group-containing compound may be a monosaccharide, a disaccharide, an oligosaccharide, a polysaccharide, a polyhydric alcohol, a sugar alcohol, a polyphenol, or a polyvinyl alcohol. These hydroxyl group-containing compounds may be commercially available or may be synthesized by a known method.

[0058] The content of the hydroxyl group-containing compound in the second agent may be appropriately adjusted according to the content of the boronic acid group in the first agent. For example, the hydroxyl group in the hydroxyl group-containing compound is preferably 0.1 to 4 equivalents, more preferably 0.5 to 3.5 equivalents, even more preferably 1.0 to 3.0 equivalents, and particularly preferably 1.5 to 2.5 equivalents, relative to 1 equivalent of the boronic acid group in the gelatin derivative in the first agent. In the adhesive of this embodiment, by determining the content of the hydroxyl group-containing compound in the second agent so that the boronic acid group and the hydroxyl group in the gelatin derivative are in the above ratio, the gelatin derivative can be sufficiently crosslinked, and the adhesive strength and pressure resistance can be further increased.

[0059] The hydroxyl-containing compound may be composed of only one type of hydroxyl-containing compound, or may be a mixture of two or more types of hydroxyl-containing compounds. When the second agent contains two or more types of hydroxyl-containing compounds, it is preferable that the total content is within the above numerical range.

[0060] <Solvent> The second agent may contain a solvent. The solvent is preferably an aqueous solvent. As the aqueous solution, those already explained as the aqueous solution that may be contained in the first agent can be used.

[0061] Among them, a phosphate buffer solution having a pH of 3 to 8 is preferred, and a phosphate buffer solution having a pH of 4 to 6 is more preferred. It is preferred that the ionic strengths of both aqueous solvents are adjusted so that the pH is about 8 to about 10 when the first agent containing a solvent and the second agent containing a solvent are mixed in the same volume. For example, the pH can be set in the above range when mixed in the same volume by using a carbonate buffer solution having a pH of 10 and an ionic strength of 0.05 to 0.1 as the first agent and a phosphate buffer solution having a pH of 4 and an ionic strength of 0.01 to 0.03 as the second agent. Alternatively, the first agent may be a carbonate buffer solution having a pH of 10 and an ionic strength of 0.05 to 0.1, and the second agent may be a phosphate buffer solution having a pH of 4 and an ionic strength of 0.01 to 0.07.

[0062] <Additives> The second agent may be composed of only a hydroxyl group-containing compound, or may be composed of only a hydroxyl group-containing compound and a solvent, or may contain other additives within the scope of the effect of the present embodiment. Examples of additives include the additives described as those that may be contained in the first agent, and the same applies to preferred embodiments.

[0063] [Adhesive manufacturing method] The adhesive can be obtained by separately preparing the first and second agents. The first agent can be produced, for example, by mixing a gelatin derivative and, if necessary, a solvent and additives, etc., by a known method. The second agent can be produced, for example, by mixing a hydroxyl group-containing compound and, if necessary, a solvent and additives, etc., by a known method.

[0064] The obtained first and second agents can be filled in a predetermined container, such as a dispenser made of plastic such as polypropylene. When used as a tissue adhesive, it is preferable to fill one side of a double syringe type dispenser, which is used when applying to tissue and can mix both agents at the tip, with the aqueous solution of the first agent, and the other side with the aqueous solution of the second agent.

[0065] In the first and / or second agent, the gelatin derivative represented by formula (1) and / or formula (2) may form colloidal particles in the solvent. In this case, it is preferable that the first and / or second agent does not form a coacervate but forms a uniformly dispersed phase of colloidal particles containing the gelatin derivative. If the first and / or second agent forms a coacervate, the curing reaction may be non-uniform. On the other hand, if the colloidal particles are uniformly dispersed in the solvent, the curing reaction proceeds uniformly, thereby obtaining a stronger adhesive strength.

[0066] [How it applies to your organization] The adhesive can be applied to incisions and skin wounds in various surgical procedures such as digestive surgery, respiratory surgery, cardiovascular surgery, neurosurgery, and oral surgery.

[0067] By mixing the two parts, a curing reaction immediately occurs to form a cured product. The mixing ratio of the first part and the second part may be appropriately adjusted depending on the content of amino groups in the first part and the content of hydroxyl group-containing compounds in the second part. The temperature during the curing reaction is not particularly limited, but is generally preferably 15 to 45°C, more preferably 20 to 40°C. The curing time is not particularly limited, but sufficient adhesive strength can be obtained in 1 to 30 minutes.

[0068] This adhesive can be used as a wound dressing for covering wounds in biological tissues, as an adhesion inhibitor for preventing postoperative adhesions, and as a hemostatic material for stopping bleeding from biological tissues.

[0069] In addition, since the cured product of this adhesive has both excellent tissue adhesion and flexibility in an aqueous environment containing blood and exudates, it can be used as a sealant to stop leakage of contents from a colon anastomosis, for example, by applying it to a colon anastomosis that involves bleeding. In addition, this adhesive can also be used as a filler.

[0070] The method of use is not particularly limited, but a preferred mode is to apply the composition to the target site (tissue) using a sprayer and allow a hardened product (gel) to form on the tissue. EXAMPLES

[0071] The present invention will be described below with reference to examples, but the present invention is not limited to these.

[0072] [Example 1] In Example 1, an adhesive consisting of a first part and a second part was produced by the method described below.

[0073] (1) Synthesis of gelatin derivatives A gelatin derivative (hereinafter, appropriately referred to as "PBA-ApGltn") having a phenylboronic acid group introduced therein was synthesized using raw gelatin (gelatin derived from Alaska pollack, hereinafter, appropriately referred to as "Org-ApGltn"). In this example, as shown in Figure 2, PBA-ApGltn was obtained by reacting 4-formylphenylboronic acid with the amino group of Org-ApGltn to form a Schiff base, and then reducing the resulting Schiff base to a stable secondary amine with a reducing agent.

[0074] 60 g of gelatin derived from Alaska pollock (Org-ApGltn) (Nitta Gelatin Co., Ltd., weight average molecular weight (Mw): 85 kDa, amino group content: 324 μmol / g) was dissolved in 800 mL of ultrapure water, and while stirring at 55 °C, 200 equivalents of 4-formylphenylboronic acid (Tokyo Chemical Industry Co., Ltd.) was added to the solution together with ethanol (Junsei Kagaku Co., Ltd.) per 100 equivalents of the amino group amount (324 μmol / g) of Org-ApGltn to form an imine bond between 4-formylphenylboronic acid and the amino group of Org-ApGltn. After stirring at the same temperature for 1 hour, 2-picoline borane (Junsei Kagaku Co., Ltd., 58.3 mmol) was added to the mixture together with ethanol to reduce the imine. The resulting mixed solution (PBA-ApGltn concentration: 5 mass / volume%, water:ethanol = 420 mL:180 mL) was stirred at 55 °C for 17 hours to allow the reaction to proceed. Then, the reaction solution (600 mL) was dropped into 500 mL of a mixed solution of ethanol:ethyl acetate (250 mL:250 mL) to purify PBA-ApGltn. The resulting reprecipitate was washed with 300 mL of ethanol (1 hour x 3 times) to remove unreacted 4-formylphenylboronic acid and 2-picoline borane. It was then dried under reduced pressure for 3 days to obtain PBA-ApGltn with a yield of 99.1 mass%.

[0075] The obtained PBA-ApGltn was analyzed by Fourier transform infrared spectroscopy (FT-IR) at 3285 cm -1 The increase in the secondary amine peak was observed, and the proton nuclear magnetic resonance ( 1H-NMR showed a peak of the aromatic ring derived from the phenylboronic acid group at 7.5 to 8.0 ppm. From these qualitative analysis results, it was confirmed that the phenylboronic acid group was introduced into PBA-ApGltn. In other words, PBA-ApGltn contains a structure represented by the following formula (3).

[0076] [ka]

[0077] In addition, the boronic acid group introduction rate (hereinafter referred to as "DS") of PBA-ApGltn was calculated from the values ​​obtained by quantifying the number of amino groups in the raw gelatin and the number of amino groups in Cat-ApGltn using the 2,4,6-trinitrobenzenesulfonic acid method (TNBS method). The DS was 58 mol%. The gelatin derivative of this example with a DS of 58 mol% is referred to as "58PBA-ApGltn".

[0078] (2) Synthesis of hydroxyl-containing compounds A hydroxyl-containing compound (hereinafter, referred to as "Cat-ApGltn") was synthesized by introducing a catechol group having a hydroxyl group into raw gelatin (gelatin derived from Alaska pollack, hereinafter, referred to as "Org-ApGltn" as appropriate) using the raw gelatin. In this example, as shown in Figure 3, Cat-ApGltn was obtained by reacting 3,4-dihydroxybenzaldehyde with the amino group of Org-ApGltn to form a Schiff base, and then reducing the resulting Schiff base to a stable secondary amine with a reducing agent.

[0079] 60 g of gelatin derived from Alaska pollock (Org-ApGltn) (Nitta Gelatin Co., Ltd., weight average molecular weight (Mw): 85 kDa, amino group content: 324 μmol / g) was dissolved in 700 mL of ultrapure water, and while stirring at 55 °C, 500 equivalents of 3,4-dihydroxybenzaldehyde (Tokyo Chemical Industry Co., Ltd.) was added to the solution together with ethanol (Junsei Kagaku Co., Ltd.) per 100 equivalents of the amino group amount (324 μmol / g) of Org-ApGltn to form an imine bond between 3,4-dihydroxybenzaldehyde and the amino group of Org-ApGltn. After stirring at the same temperature for 1 hour, 2-picoline borane (Junsei Kagaku Co., Ltd., 145.8 mmol) was added to the mixture together with ethanol to reduce the imine. The resulting mixed solution (Org-ApGltn concentration: 6 mass / volume%, water:ethanol = 105:45 mL) was stirred at 50 °C for 17 hours to allow the reaction to proceed. Then, the reaction solution (150 mL) was dropped into 8000 mL of cold ethanol (-7 to 4 °C) to purify Cat-ApGltn. The resulting reprecipitate was washed with 8000 mL of ethanol (1 hour x 2 times) to remove unreacted 3,4-dihydroxybenzaldehyde and 2-picoline borane. It was then vacuum dried for 3 days to obtain Cat-ApGltn in a yield of 96.7 mass%.

[0080] The obtained Cat-ApGltn was analyzed by Fourier transform infrared spectroscopy (FT-IR) at 3285 cm -1 The increase in the secondary amine peak was observed, and the proton nuclear magnetic resonance ( 1 H-NMR showed a peak of aromatic ring derived from catechol group at 6.8 to 7.0 ppm. From these qualitative analysis results, it was confirmed that catechol group was introduced into Cat-ApGltn. That is, Cat-ApGltn contains a structure represented by the following formula (4).

[0081] [ka]

[0082] In addition, the catechol group introduction rate (hereinafter, appropriately referred to as "DS") of Cat-ApGltn was calculated from the values ​​obtained by quantifying the number of amino groups in the raw gelatin and the number of amino groups in Cat-ApGltn using the 2,4,6-trinitrobenzenesulfonic acid method (TNBS method). The DS was 51 mol%. The hydroxyl group-containing compound of this example with a DS of 51 mol% is referred to as "51Cat-ApGltn".

[0083] (3) Preparation of the first agent 58PBA-ApGltn (gelatin derivative) was dissolved in 0.1 M phosphate buffer at a concentration of 10 w / v % to prepare the first agent. The pH of the first agent was set to pH 6.

[0084] (4) Preparation of the second agent 51Cat-ApGltn (a hydroxyl group-containing compound) was dissolved in 0.1 M phosphate buffer (pH 4) at a concentration of 10 w / v % to prepare a second agent.

[0085] [Example 2] In Example 2, the pH of the first part was set to pH 8. Except for that, an adhesive consisting of the first part and the second part was produced in the same manner as in Example 1.

[0086] [Example 3] In Example 3, a 0.1 M carbonate buffer solution was used as the solvent for the first part, and the pH of the first part was set to pH 10. Except for this, an adhesive consisting of the first part and the second part was produced in the same manner as in Example 1.

[0087] [evaluation] 1. Compression strength test According to ASTM F2392-04, the sealing strength (compression resistance strength) was measured using a porcine large intestine (φ30 mm) as a substrate (see FIG. 4). 100 μL each of the first and second parts of the adhesives produced in Examples 1 to 3 were mixed and applied to the porcine large intestine to prepare a cured adhesive product with a thickness of 1.0 mm and a diameter of 15 mm. After application, the product was left to stand at 25° C. for 10 minutes, and then 37° C. saline was poured at 2 mL / min to measure the pressure at the time of rupture. The results are shown in Table 1.

[0088] [Table 1]

[0089] As shown in Table 1, the pressure resistance of the adhesive of the examples increased as the pH of the PBA-ApGltn solution increased.

[0090] 2. Underwater adhesive strength test The adhesive strength between the adhesive prepared in Example 3 and the porcine large intestine adventitia tissue was measured by the following method. The test method was performed according to the standard (ASTM F-2258-05) of the American Society for Testing and Materials. A 2.5 cm square piece of porcine large intestine adventitia tissue was prepared, and the piece was fixed to the upper jig of the test device and the lower part of an acrylic box (10 cm x 10 cm x 10 cm) using instant adhesive. The adhesive was applied to the piece of porcine large intestine adventitia tissue fixed to the lower part of the acrylic box to a thickness of 1 mm and left to stand for 10 minutes. Thereafter, the acrylic box was filled with 500 mL of phosphate buffer (pH 7.4, 37°C), and the piece of porcine large intestine adventitia tissue fixed to the upper part of the jig was pressed at 2 N for 3 minutes, after which the adhesive strength in water was measured.

[0091] For comparison, the upper and lower pieces of porcine colon adventitia tissue were directly bonded together without using an adhesive, and a similar pressure resistance test was performed (Comparative Example 1). A similar pressure resistance test was also performed on a commercially available fibrin adhesive (CSL Behring, Beriplast (registered trademark)) (Comparative Example 2). The results are shown in Table 2.

[0092] [Table 2]

[0093] According to Table 2, it was found that the adhesive of Example 3 had superior adhesive strength to porcine colon tissue in water compared to the powders of Comparative Example 1 and Example 2.

[0094] 3. Cytotoxicity test and biocompatibility evaluation 500 μL of the first agent (PBA-ApGltn solution) and 500 μL of the second agent (Cat-ApGltn solution) of the adhesive of Example 3 were mixed in a 15 mL centrifuge tube to form a gel, 10 mL of medium was added to the resulting gel and incubated for 24 hours, and 100 μL of the resulting supernatant was added to Caco-2 cells and cultured for 24 hours. After that, the number of cells was counted using WST-8.

[0095] The results of the cytotoxicity test showed that the cells showed a survival rate of 75% or more compared to the control (sample cultured without adding supernatant), confirming low cytotoxicity.

[0096] As a result of evaluating biocompatibility by subcutaneous implantation into rats, it was confirmed that the cured adhesive (gel) of Example 3 did not cause excessive inflammatory reactions after any period of observation, and thus had high biocompatibility. [Industrial Applicability]

[0097] The adhesive of the present invention has both high biocompatibility and adhesive strength, and can be used for adhering biological tissues in an environment where bleeding or exudate is present, such as after colon surgery.

Claims

1. An adhesive comprising: A first agent containing a gelatin derivative having a structure represented by the following formula (1), and a second agent containing a hydroxyl group-containing compound. 【Chemistry 1】 In formula (1), Gltn represents a gelatin residue, X represents one selected from the group consisting of -C(O)-NH-, -NH-C(O)-, and -NH-, and L represents a single bond or a divalent linking group.

2. The adhesive according to claim 1, wherein in formula (1), the divalent linking group includes at least one selected from the group consisting of an alkylene group having 1 to 10 carbon atoms, a phenylene group, and a polyethylene oxide having 1 to 10 repeating numbers.

3. The adhesive according to claim 1 or 2, wherein the hydroxyl group-containing compound is a compound having at least two hydroxyl groups.

4. The adhesive according to any one of claims 1 to 3, wherein the structure of the hydroxyl group-containing compound is a structure represented by the following formula (2): 【Chemistry 2】 In formula (2), Gltn represents a gelatin residue, Y represents one selected from the group consisting of -C(O)-NH-, -NH-C(O)-, and -NH-, L １ represents a single bond or a divalent linking group, and n is 1 to 5.

5. The adhesive according to claim 4, wherein in formula (2), the divalent linking group is at least one selected from the group consisting of an alkylene group having 1 to 10 carbon atoms and a polyethylene oxide having 1 to 10 repeating numbers.

6. In formula (2), the hydroxyl group in the benzene ring is L １ The adhesive according to claim 4 or 5, wherein the bonding position of

7. The adhesive according to any one of claims 1 to 3, wherein the hydroxyl group-containing compound is at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, polysaccharides, polyhydric alcohols, sugar alcohols, polyphenols, and polyvinyl alcohols.

8. A wound dressing comprising an adhesive according to any one of claims 1 to 7.

9. An anti-adhesion material comprising the adhesive according to any one of claims 1 to 7.

10. A hemostatic material comprising the adhesive according to any one of claims 1 to 7.

11. A sealant comprising the adhesive according to any one of claims 1 to 7.

12. A filler comprising the adhesive of any one of claims 1 to 7.

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