JP2008104612A - Two-component adhesive - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a two-component adhesive which has improved operability compared with a conventional one and provides a sufficient adhesion strength and flexibility suitable to adhesion of biological tissues. <P>SOLUTION: The two-component adhesive is composed of a main agent and a cross-linking agent. The main component of the main agent is an albumin and a polyhydric phenol compound or a polyphenol compound, and the main component of the cross-linking agent is an aldehyde. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a two-component adhesive for living tissue. More specifically, it has excellent flexibility and adhesion performance, and is bioabsorbable consisting of albumin, polyhydric phenol compound and / or polyphenol compound, and optionally a main agent composed of additives and a crosslinking agent containing sugar aldehyde and / or alkylene aldehyde. And a two-component adhesive for living tissue having excellent safety.

  Various surgical tissue adhesives are used in medical applications. Of these, cyanoacrylates, gelatin-formaldehyde compositions and fibrin-based adhesives have received the most attention. For example, fibrin (a blood clotting bio-derived material) and gelatin-formaldehyde have each been used for surgical adhesive applications as well as cyanoacrylate. These adhesives are used only for specific applications because they have drawbacks as shown below.

Cyanoacrylates have been studied for surgical use. For example, certain isobutyl cyanoacrylate formulations have been approved for use in livestock treatment. Typically, a monomer and / or mixture of monomers is applied to the adhesion site and rapidly polymerizes into a solid. One drawback of cyanoacrylate adhesives is that the affected area needs to be dry. The other is that the solid obtained by polymerization is not absorbed into the body, and for these reasons, the usefulness of this adhesive is reduced when used in vivo. In addition, it has been reported that polymerization is exothermic and the tissue compatibility is poor. (Refer nonpatent literature 1-3.).
Bonutti, P. et al., Clinical Orthopedics and Related Research, Vol. 229, pages 241-248 (1988) Nelson, R. et al., Arch. Surg. Vol, 100, pages 295-298 (1970) Celik, H., et al., “Nonsuture closure of arterial defect by vein graft using isobutyl-2-cyanoacrylates as a tissue adhesive”, Journal of Nearosurgical Sciences Vol. 35, No. 2 (April-June 1991)

Adhesives crosslinked with formaldehyde and based on gelatin have been used experimentally since 1964, mainly in Europe. Of the proposed formulations, “GRF adhesives” (gelatin, resorcin, formaldehyde) are well known. A heated solution of gelatin is mixed in situ with a hardened product from a formaldehyde-based solution. The mixture quickly solidifies and adheres to the tissue. For GRF adhesives, the gelatin used as the main agent should be applied under warming, for example at a temperature above body temperature. Furthermore, the strength is not sufficient from the viewpoint of tissue adhesion, and there is a risk of tissue peeling. Mixing and application techniques have not been strictly determined under the clinical practice of GRF. (Refer nonpatent literature 4-7.).
Fabiani, et al., Ann. Thorac. Surg. 143-145 (1990) Fabiani, et al., Supplement J. Circulation Vol. 80, No. 3 (September 1989) Bachet, et al., J. Thorac. Cardiovasc. Surg. Vol. 83 (1982) Braunwald, et al., Surgery Vol. 59, No. 6 (June 1966)

Fibrin glue utilizes a blood clotting system to produce an adhesive or sealant composition. One commercially available composition is “Tussicol®” available from Rugis, France. The other is the “Fibrin Sealant Kit 1.0” available from Osterreichisches Institut for Hasmoderirate, GMBH (Immuno GA, A-1220, Vienna, Austria). When fibrin glue is used, blood aggregates are formed by two types of components. One of the components is a solution of fibrinogen and blood clotting factors, such as factor XIII, and the other is a solution of thrombin and calcium ions. The disadvantages of fibrin adhesives are their tensile strength (generally 50 g / cm ² or less) and slow cure time. There is also the risk of patient infection when using blood products (fibrinogen and cofactors) from multiple human donors. Therefore, a method using autologous blood for preparing a fibrin sealant has been proposed. (See Patent Document 1).
JP-A-57-149229

An adhesive composition for living tissue is known in which gelatin, which is a biological material, is used as a main agent and glutaraldehyde or the like is used as a crosslinking agent. (See Patent Document 2). However, since gelatin is contained as a main ingredient, there is a problem that it cannot be used unless it is heated. As a result of the special preparation required in the operative field, the application takes time, and there is a risk that the burden on the patient increases as the operation time is increased.
Japanese Patent Laid-Open No. 6-70972

  For these reasons, there has not been established a technique for a medical adhesive having flexibility and adhesive strength using a main agent that can be used at room temperature. These techniques are particularly useful in optimum composition in the case where the main agent and the crosslinking agent are automatically mixed and applied by a device.

As an adhesive that does not require warming and dissolving operation of the main agent, an adhesive for biological tissue of a genetic manipulation-derived protein and a polyfunctional aldehyde has been proposed. (See Patent Document 3). However, since only the protein is used as the main agent, the physical properties after gelation are hard and brittle, so there is a risk of vascular infarction and rebleeding due to film peeling.
JP 2000-288079 A

In addition, an adhesive for living tissue using a collagen protein partial hydrolyzate and a water-soluble chitin derivative as a main agent and an alkylene aldehyde as a crosslinking agent is known. (See Patent Document 4). Since the water-soluble chitin derivative as an additive is extremely difficult to impart sufficient flexibility to the gelled product, there is a concern similar to the above.
JP 2000-290633 A

Furthermore, an adhesive sealant composition using a protein within a specific pH range as a main ingredient and a modified polyethylene glycol as a cross-linking agent is known. (See Patent Document 5). However, since the usable pH is limited, its versatility is limited.
Japanese Patent No. 3592718

In addition, an adhesive composition using plasma protein as a main ingredient and a polyfunctional aldehyde as a crosslinking agent is known. (See Patent Document 6). In this patent document, examples using aldehydes derived from trehalose are disclosed. However, since trehalose aldehyde alone is used as a cross-linking agent, it takes 1 hour to cure the adhesive, and the tear strength after 1 hour is only 245 g / cm ² at the most. Not practical as an agent.
Japanese Patent No. 3483882

  An object of the present invention is to provide a two-component adhesive that not only improves operability, but also provides sufficient adhesive strength and flexibility that is compatible with living tissue adhesion.

The present inventors use a two-component adhesive composed of a main component mainly composed of albumin and a polyphenol compound and / or a polyphenol compound, and a cross-linking agent mainly composed of an aldehyde. The present invention has been completed by finding that it can be easily applied without taking any form of use, and can provide sufficient adhesive strength and flexibility. That is, the present invention has the following configuration.
The present invention is a two-component adhesive comprising a main agent and a cross-linking agent, wherein the main agent is albumin and a polyhydric phenol compound and / or a polyphenol compound as main components, and the cross-linking agent is an aldehyde as a main component. It is an adhesive.
In the present invention, the two-component adhesive preferably contains an additive in the main agent.
In the present invention, the additive is preferably a sodium salt of an organic acid.
Moreover, it is preferable that this additive is sodium benzoate.
Moreover, it is preferable that this polyhydric phenol compound is resorcinol or β-resorcylaldehyde (2,4-dihydroxybenzaldehyde).
The polyphenol compound is preferably a flavonoid.
Moreover, it is preferable that the polyphenol compound or polyphenol compound content in a main ingredient is 0.01 to 20 weight%.
Moreover, it is preferable that the additive content in a main ingredient is 0.001 to 20 weight%.
The albumin preferably contains at least one albumin selected from the group consisting of human blood-derived albumin, bovine blood-derived albumin main fragment, and chemically modified albumin.
Moreover, it is preferable that the albumin content in a main ingredient is 5-45 weight%.
Moreover, it is preferable that this aldehyde consists of alkylene aldehyde and / or sugar aldehyde.
The alkylene aldehyde is preferably composed of one or more alkylene aldehydes selected from the group consisting of formaldehyde, glyoxal, succinaldehyde, glutaraldehyde, and malealdehyde.
Moreover, it is preferable that the alkylene aldehyde content in a crosslinking agent is 0 to 30 weight%.
Moreover, it is preferable that this sugar aldehyde is obtained by oxidizing sugars having 1 to 1,000 repeating units.
The sugar aldehyde is preferably at least one selected from those obtained by oxidizing monosaccharides, disaccharides, trisaccharides and tetrasaccharides.
Moreover, it is preferable that the sugar aldehyde content in a crosslinking agent is 0 to 30 weight%.
Moreover, it is preferable that the aldehyde content in a crosslinking agent is 0.001-30 weight%.
Moreover, it is preferable to mix a main ingredient and a crosslinking agent by the weight ratio of 100: 1-100: 100.
The two-component adhesive is preferably used for hemostasis or biological tissue.

  Compared to conventional adhesives, the two-part adhesive of the present invention not only eliminates the need for preheating of the main agent, but also provides a material with sufficient adhesive strength and flexibility to fit the tissue. be able to. By using albumin as the main agent, prior heating in the main agent using gelatin of the prior art becomes unnecessary, and furthermore, when attaching a living tissue using a two-component mixing device that enables precise mixing of the main agent and the crosslinking agent. It is possible to provide a material that is particularly effective for optimizing the bonding time. In addition, a material capable of freely controlling sufficient strength and appropriate flexibility can be provided that surpasses the prior art.

  In the present invention, gelation means that the molecular weight becomes infinite due to a polymerization reaction between a biological substance as a main ingredient and an aldehyde as a cross-linking agent, resulting in a swollen body that is insoluble in any solvent. This is called chemical reaction.

In medicine and biology, aldehyde aqueous solution is an antiseptic for the preparation of biological tissues,
1161740699468_0
Widely used in This is because the aldehyde penetrates into and out of the tissue and binds mainly to the protein in the tissue, thereby damaging the three-dimensional structure of the protein and inhibiting various biological activities such as enzyme activity, transport and secretion. It is thought to be because. Since ancient times, aldehydes have been used to immobilize biological tissues, and in particular, treatment with glutaraldehyde, an alkylene aldehyde, has been used as a preparation stage for specimen preparation for microscopic observation. Alkylene aldehyde represented by glutaraldehyde has a very high reaction rate. The use of alkylene aldehydes is preferred in order to show high reactivity in aqueous solutions and to obtain adhesion times on the order of minutes.

  Formaldehyde and glutaraldehyde are used as a crosslinking agent for the above-mentioned GRF adhesive, which is an adhesive for living tissue, but in recent years their biotoxicity is a concern. This is because the aldehyde group of alkylene aldehyde reacts with proteins such as enzymes that make up the living body and there is a risk of introducing an alkylene group = hydrophobic group into the molecule, losing enzyme activity, etc. If unreacted alkylene aldehyde remains This is because, as mentioned above, there is a risk of causing toxicity that inhibits life activity. That is, when using a biological tissue adhesive together with a surgical suture, it is preferable from the viewpoint of shortening the operation time and reducing the burden on the patient that the gelation time is as short as possible, but the alkylene aldehydes are as described above. In addition, the remaining unreacted material has a problem in terms of biological safety. Therefore, as a result of intensive investigations on improving the safety of adhesives for biological tissues, the present inventors have found that unreacted substances remain and protein molecules should be used if saccharides that are indispensable for human life activities can be aldehyded. Even if it reacted, it was thought that the influence could be mitigated to the maximum by the hydrophilic structure of sugar. Moreover, when aldehyde-ized saccharide | sugar was examined, it discovered that the ring structure of saccharide | sugar was simple and could be obtained with a sufficient yield.

  The GRF adhesive described above is a two-component adhesive in which the main ingredients are gelatin and resorcin, and the crosslinking agent is formaldehyde and glutaraldehyde. The adhesion mechanism is such that the amino group of gelatin and the aldehyde undergo gelation as the polycondensation reaction proceeds with the elimination of water. Here, resorcin is to impart flexibility to the adhesive, and in the case of a GRF adhesive, the amount is about one third of that of gelatin. The polymerization rate can be controlled by increasing or decreasing the mixing ratio or the total concentration of the two types of aldehydes which are crosslinking agents. The physical properties of an adhesive (sometimes referred to as a gelled product) obtained by mixing two liquids depend on the characteristics of the main agent that occupies most of the composition.

  GRF adhesive has come to be widely recognized as a surgical adhesive, but as mentioned above, gelatin is a solid at room temperature, so it is necessary to warm the main agent before use to dissolve the gelatin. Have On the other hand, since the main ingredient of the present invention uses albumin instead of gelatin, the solution state can be maintained during the distribution process, storage, and use, and there is no need to heat and dissolve the main ingredient before use. There is an advantage.

As a mechanism of the gelation reaction of the GRF adhesive, the reaction of the main ingredient content by the esterification reaction between the carboxylic acid residue of gelatin and resorcinol greatly contributes to the gelation. (Refer nonpatent literature 8). In addition, as described above, in the GRF adhesive, flexibility was imparted to the gel for the first time by adding about 1/3 of resorcin to gelatin, but when albumin and resorcin are dissolved in the main agent, There was a problem that it was not possible to add resorcinol in such an amount that it could exhibit flexibility. Resorcin itself is readily soluble in water, and the reason why resorcin is poorly soluble in albumin aqueous solution is not well understood, but because the compatibility between albumin and resorcin is not good, a large amount of resorcin may be added to the main ingredient. I think I can't.
Kawasaki, N., Kato, H., Serizawa, K., GRF (gelatin-resorcin-formaldehyde) as an anti-ulcer drug sustained release base material, Tokyo Medical Journal 57 (5): 481-490, 1999

  Therefore, the present inventors have intensively studied about increasing the amount of resorcin addition to the aqueous solution of albumin, which is the main ingredient, and as a result, the addition of resorcin to the main ingredient by adding an additive capable of compatibilizing both albumin and resorcin. We thought that we could increase the amount, and we proceeded with further studies. As a result of the study, it was found that the amount of resorcin addition can be increased by using, for example, a material having a structure composed of a hydrophobic (benzene) skeleton + a hydrophilic group as an additive. The reason why the amount of resorcin addition increased by using an additive having a hydrophobic (benzene) skeleton + hydrophilic group is not well understood, but it has a surface-active effect on both albumin and resorcin. Is considered to have increased compatibility and solubility.

  Further, when chemical analysis of the main agent solution composed of albumin, resorcin, and additives was performed, it was found that the formation of ester bonds due to the reaction between albumin and resorcin did not occur. On the other hand, as described above, in the GRF adhesive, gelatin and resorcin are bonded to each other by an esterification reaction between gelatin and resorcin in the main agent. It is different from the agent. That is, the gel itself is crosslinked and cannot be analyzed. However, in the adhesive of the present invention and the conventional GRF adhesive, the gel obtained by the reaction of the main agent and the crosslinking agent is as shown in FIG. It is thought that there is a structural difference.

  In the present invention, resorcin first reacts with aldehyde before reacting with albumin to form both terminal aldehyde-modified resorcin, and then both terminal aldehyde-modified resorcin reacts with albumin. As shown, albumin binds to both ends of aldehyde-modified resorcin to form a gel. By obtaining such a reaction mechanism and an intermediate product, it is considered that not only the reactivity is improved, but also the flexibility is enhanced by incorporating an aldehyde as a spacer in the main chain structure of the gel. It was also confirmed that the safety of the two-component adhesive of the present invention is equal to or less than that of the existing technology GRF adhesive.

Hereinafter, the present invention will be described in more detail.
Examples of the additive of the present invention include formic acid, acetic acid, propionic acid, butyric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, crotonic acid, lactic acid, levulinic acid, pyrotartaric acid, oxalic acid, malonic acid, maleic acid Organic acids consisting of acids, fumaric acid, succinic acid, glutaric acid, malic acid, adipic acid, citric acid, benzoic acid, phthalic acid, isophthalic acid, terephthalic acid, salicylic acid, acetylsalicylic acid, gallic acid and / or these organic Preferably, the acid salt is at least one selected from the group of organic acid salts consisting of ammonium salts, sodium salts, potassium salts, calcium salts, iron salts, copper salts, and aluminum salts. A sodium salt is more preferable from the viewpoint of safety, handleability and cost, and sodium acetate, sodium citrate and sodium benzoate are still more preferable.

As mentioned above, when albumin and a polyphenol compound are mixed in water, the hydrophobic benzene ring of the polyphenol compound is mixed to reduce the compatibility with the hydrophilic albumin molecule. It is thought that poor dissolution (precipitation) of resorcin may occur. In order to avoid this precipitation, by using an additive having hydrophilic and hydrophobic substituents, that is, having a surface active action, it is not possible to dissolve both different properties and make a uniform aqueous solution. (See Fig. 2).
In the present invention, it is preferable to use an organic acid salt, particularly an organic acid sodium salt as an additive because the additive used can suppress the pH change of the solution to a small level and has high biological safety. The additive is sodium benzoate, which has not only a high compatibilizing effect between albumin and polyhydric phenol compounds by having a benzene ring like polyhydric phenol compounds, but also has a track record for biosafety of use in food additives. It is more preferable to use as.

(式中Ｘは−ＣＨ₃ ，−ＣＨ₂ ＯＨ又は−ＣＨＯ基を表し、ｎは０〜３の整数を表し、ｎが２又は３の場合Ｘは同一又は各々異なる基であってもよい。） In the present invention, the polyhydric phenol compound is preferably a divalent phenol compound represented by the following general formula (1). Since there is concern about a decrease in water solubility due to an increase in hydrophobic substituents, resorcin, β-resorcylaldehyde (2,4-dihydroxybenzaldehyde) or γ-resorcylaldehyde (2,6-dihydroxybenzaldehyde), 4-dihydroxy More preferred are benzyl alcohol and 2,6-dihydroxybenzyl alcohol.

(In the formula, X represents a —CH ₃ , —CH ₂ OH or —CHO group, n represents an integer of 0 to 3, and when n is 2 or 3, X may be the same or different groups. )

In the present invention, the polyphenol compound is preferably a flavonoid represented by the following general formula (2). When using flavonoids, there is a concern about the decrease in water solubility due to an increase in hydrophobic substituents, so the amount of additive having a surface active action should be increased compared to the case of using a polyphenol compound such as resorcin. Is preferred.

  In the present invention, the content of the polyphenol compound and / or polyphenol compound in the main agent is preferably 0.01 to 20% by weight, and if the content is too low, the flexibility-imparting effect is not sufficient, so 1% by weight or more. Is more preferable, and if it is too much, the solubility may be lowered, so that 18% by weight or less is more preferable.

  The additive content in the main agent of the present invention is preferably 0.001 to 20% by weight. If the amount is too large, the viscosity of the main agent is excessively increased. Therefore, the amount is more preferably 18% by weight or less, and if the amount is too small, a sufficient compatibility effect cannot be expected.

  The albumin used in the present invention is not limited in its origin, such as human serum albumin, human serum albumin (hereinafter referred to as recombinant human serum albumin) produced by gene recombination technology, bovine serum albumin and the like. In consideration of application to humans, human serum albumin or recombinant human serum albumin is preferable, and human serum albumin is more preferable from the viewpoint of easy availability.

  In the present invention, the total content of albumin and polyhydric phenol and / or polyphenol compound in the main agent is preferably 38 to 55% by weight. If it is this range, when what mixed the main ingredient and the crosslinking agent beforehand is apply | coated to an application site | part, since the viscosity of the grade which does not flow down from an application site | part can be ensured, it is preferable. If the total content is too high, the viscosity of the main agent becomes high and mixing with the crosslinking agent may be difficult. Therefore, the total content in the main agent is more preferably 50% by weight or less. If the total content in the main agent is too low, the adhesive may be cast at the application site, so 40% by weight or more is more preferable.

  In the present invention, the albumin content in the main ingredient is preferably 5 to 45% by weight. If the albumin content is too low, there is a possibility that a problem of easily flowing off from the adhesion site occurs. Therefore, the content is preferably 10% by weight or more, more preferably 18% by weight or more, and even more preferably 26% by weight or more. On the other hand, if the albumin content is too high, the fluidity is lowered and the handling property may be lowered. Therefore, the albumin content in the main ingredient is more preferably 43% by weight or less, still more preferably 39% by weight or less, and even more preferably 35% by weight or less.

  In the present invention, the aldehyde is preferably one or more aldehydes selected from alkylene aldehydes and sugar aldehydes. By combining a plurality of types of aldehydes, it becomes easy to control the gelation time and the gelation reaction rate that could not be obtained with a single aldehyde.

  In the present invention, the alkylene aldehyde is preferably at least one alkylene aldehyde selected from the group consisting of formaldehyde, glyoxal, succinaldehyde, glutaraldehyde, and malealdehyde. The combination of formaldehyde and glutaraldehyde is more preferable because the gelation time can be controlled more strictly. The combination of glyoxal and glutaraldehyde is more preferable because toxicity of formaldehyde can be reduced.

  In this invention, it is preferable that the alkylene aldehyde content in a crosslinking agent is 0 to 30 weight%. If the content is too low, a sufficient gelled product cannot be obtained, and there is a risk of re-bleeding due to early dissolution, so 0.5% by weight or more is more preferable, and 1% by weight or more is more preferable. If the content is too high, the gelation reaction is completed too early and sufficient adhesion cannot be achieved. Accordingly, the alkylene aldehyde content is more preferably 25% by weight or less, further preferably 20% by weight or less, and further preferably 15% by weight or less.

  In the present invention, the sugar chain part of the sugar aldehyde preferably has 1 to 1000 repeating units. More preferably, it is 1-100, More preferably, it is 1-10, More preferably, it is 1-4. Specifically, glucose, galactose, mannose, fucose, xylose, sucrose, lactose, maltose, isomaltose, cellobiose, trehalose, gentiobiose, laminaribiose, chitobiose, xylobiose, inulobiose, mannobiose, hyarobiouronic acid, chondrocin, cellobiose, Examples include raffinose and neuramin lactose. If the repeating unit is too large, the reactivity may decrease. The disaccharide trehalose has a function of protecting cells and intracellular substances when frozen or dried, and is a substance that is expected to play a role as a storage and stabilizer in the fields of medicine, cosmetics, food, etc. Therefore, it is preferable as a raw material.

  In the present invention, the method for preparing the sugar aldehyde is not limited, but can be obtained by, for example, oxidative decomposition of the sugar. Examples of oxidizing agents include chlorine, hypochlorous acid, hydrogen peroxide, potassium permanganate, ammonium persulfate, periodic acid, nitric acid, and ozone. It is preferable to use periodic acid and its salt from the point which can be performed.

  In the present invention, the sugar aldehyde content in the crosslinking agent is preferably 0 to 30% by weight. If the sugar aldehyde content is too high, the gelation reaction may be completed too early and sufficient adhesion may not be achieved. Therefore, the content is preferably 25% by weight or less, more preferably 20% by weight or less, and further preferably 15% by weight or less. More preferred. If the content is too low, the gelation reaction may not be involved, so 0.5% by weight or more is more preferable, and 1% by weight or more is more preferable.

  In the present invention, the aldehyde content in the crosslinking agent is preferably 0.001 to 30% by weight. If the aldehyde content is too low, sufficient adhesive strength may not be obtained, so 0.01% by weight or more is more preferable, 0.1% by weight or more is more preferable, and 0.5% by weight or more is even more preferable. If the content is too large, the gelation reaction may be completed too early and sufficient adhesion may not be achieved. Therefore, the content is preferably 28% by weight or less, more preferably 25% by weight or less, and even more preferably 20% by weight or less.

  In the present invention, the aldehyde contained in the cross-linking agent reacts with the main protein to cause gelation. The gelation rate can be controlled by increasing or decreasing the concentration of the aldehyde. The speed of gelation depends greatly on the reactivity of the aldehyde used. For example, when glutaraldehyde, which is an alkylene aldehyde, is used, the reaction can be completed quickly because it is a low molecular weight and bifunctional aldehyde. When formaldehyde is used, the reaction rate is slow because the structure has only one aldehyde group, and the reaction proceeds slowly. Furthermore, when trehalose aldehyde is used, the reaction is slow because the molecular weight is larger than that of normal alkylene aldehyde, but since it is a bifunctional aldehyde, a crosslinking point can be reliably formed, so gelation with time will occur. There is an advantage that a gel that proceeds reliably and has a sufficient reaction rate and high water resistance can be obtained.

  In the present invention, the properties of the adhesive gelled product obtained by mixing the main agent and the crosslinking agent largely depend on the properties of the main agent, but the influence of the aldehyde used as the crosslinking agent on the properties of the adhesive gelled product is not zero. That is, when an aldehyde having high reactivity is used, the gelation reaction is completed quickly and a stronger gel is produced. On the other hand, if the reactivity of aldehyde is low and gelation is slow, a softer gel is produced. As described above, the aldehyde used may be appropriately selected in consideration of the use as an adhesive, the application site, the bonding time, the bonding strength, the flexibility, the safety, and the like, and is not limited at all. For example, when used for hemostasis at the time of surgery or adhesion to an affected area, it is preferable that the time required for adhesion is somewhat short, the adhesive strength and flexibility are excellent, and the safety of unreacted aldehyde is high. In this case, specifically, sugar aldehyde / alkylene aldehyde = 0 to 15 wt% / 0.001 to 15 wt% is preferably used.

  In the present invention, the gelation completion time is preferably 30 seconds or more and less than 120 seconds. 35 seconds or more and less than 110 seconds are more preferable, and 40 seconds or more and less than 100 seconds are more preferable. If the gelation completion time is too early, gelation proceeds before application to the application site, so that the adhesive force may be reduced or disappear. If the gelation completion time is too late, it takes a long time to adhere, so that it may flow down from the application site or the operation time may be delayed.

  In the present invention, the gelation reaction rate is preferably 50% or more. 55% or more is more preferable, and 60% or more is more preferable. When the gelation reaction rate is too low, the mixture of the main agent and the crosslinking agent dissolves rapidly due to moisture in the living body, and as a result, rebleeding from the application site may occur. By extracting the gel with physiological saline or the like, the moisture-insoluble component = the complete gel weight can be determined.

  In the present invention, it is preferable to mix the main agent and the crosslinking agent in a weight ratio of 100: 1 to 100: 100. If the crosslinking agent ratio is too small, a sufficient gelled product may not be obtained. Therefore, 100: 5 or more is more preferable. If the crosslinking agent ratio is too large, the crosslinking agent viscosity is low. 100: 70 or less is more preferable because the viscosity of the mixture decreases and the mixture may flow down from the application site until the gelation is completed, and a sufficient adhesive force may not be obtained.

  The two-component adhesive of the present invention can exhibit the flexibility and adhesive strength required for hemostasis and biological tissue adhesion, and therefore can be used as an adhesive for hemostasis or biological tissue. . At that time, by using a device for a two-component adhesive capable of extruding the main agent and the cross-linking agent at an accurate weight ratio, it becomes possible to mix an appropriate amount of the main agent and the cross-linking agent. It is preferable because the residual can be suppressed.

  In the present invention, the content of albumin / (polyhydric phenol compound or polyphenol compound) / additive / water in the main ingredient is a weight ratio of 20 to 45/1 to 15 / 0.1 to 20/20 to 78.9. It is preferable. If the weight ratio of albumin, polyphenol compound or polyphenol compound, and additive is too large, the fluidity of the viscosity of the main agent will be reduced, which may cause problems in use. If it is too small, it will be applied before gelation. There is a risk of spilling from the site. Therefore, the weight ratio of 21-44 / 2-14 / 0.2-19 / 23-76.8 is more preferable, and 22-43 / 3-13 / 0.3-18 / 26-74.7 is still more preferable. .

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited by these.

(Measurement of gel completion time)
This was carried out using a dynamic viscoelasticity measuring machine (RS1 manufactured by HAAKE). A disposable parallel plate sensor system was used under the conditions of stress: 100 Pa, frequency: 1 Hz, measurement temperature: 20 ° C., gap: 1 mm. The measurement sample was prepared as follows. The main agent having the concentration shown in the Examples; 0.2 g was added onto the parallel plate, and the crosslinking agent; 0.02 g was added in an amount shown in the Examples. The measurement was started immediately after mixing it with a spatula for 5 seconds. The gelation time was evaluated by the position where the elastic term (G ′) and the viscous term (G ″) intersect (G ′ = G ″), that is, the gel point. A gel point of 30 seconds or more and less than 120 seconds was judged good, and a gel point of less than 30 seconds and 120 seconds or more were judged bad.

(Measurement of gelation reaction rate)
The main agent having the concentration shown in the Examples: 1 g and a crosslinking agent: 0.1 g were added into a 30 mL glass vial, and mixing was continued for 10 minutes with a spatula. 20 mg of physiological saline was added to the gel obtained by mixing, and incubation was performed at 37 ° C. with shaking for 24 hours. The gel after the incubation was taken out and frozen at −5 ° C. for 24 hours, and freeze-dried for 48 hours in a vacuum dryer. The gelation reaction rate was calculated by dividing the dry weight after lyophilization by the sum of the theoretical dry weight of the main agent and the crosslinking agent before mixing. When the gelation reaction rate was 50% by weight or more, it was regarded as good, and when it was less than 50% by weight, it was regarded as poor.

(Measurement of main agent viscosity)
This was carried out using a dynamic viscoelasticity measuring machine (RS1 manufactured by HAAKE). Using a disposable parallel plate sensor system, the rotation was controlled under the conditions of a rotation speed of 10 (1 / s), a measurement temperature: 20 ° C., and a gap: 1 mm. The measurement sample was prepared as follows. The measurement was started by adding 0.2 g of the main agent having the concentration shown in the Examples on the parallel plate. The viscosity after 180 seconds after the start of measurement was recorded as the main agent viscosity, and a viscosity of 20 or more and less than 80 Pa · s was regarded as good, and other viscosities were regarded as poor. If the viscosity is less than 20 Pa · s, there is a risk of outflow from the tissue, which is not preferable. If the viscosity is 80 Pa · s or more, mixing with a crosslinking agent becomes difficult due to high viscosity, and gelation may occur unevenly. is there.

(Tensile strength measurement)
In a plastic petri dish having a diameter of 60 mm, 0.1 g of the main agent having the concentration shown in the examples and then 0.01 g of the crosslinking agent were added in the amounts shown in the examples, and after mixing for 5 seconds, a wool specimen (0.3 × 10 × 50 mm) ) Was applied within the range of 10 × 10 mm and bonded to the tip of another test piece, and then pressed with a 50 g weight for 2 minutes. Thereafter, evaluation was performed using a tensile tester. The tensile tester was an Autograph AGS-50NJ (Shimadzu Corporation). The test conditions are as follows. The load cell was 5 kg, the crosshead speed was 30 mm / min, and the distance between chucks was 7 cm. Average values (n = 5) were used as data. The maximum test force was the tensile strength, and 1.0 to 2.0 kgf was good. The elastic modulus was calculated from the slope of a tangent line connecting two points of 0.2 and 0.4 kgf, and an elastic modulus of 110 kgf / mm ² or less was considered good.

(Synthesis of trehalose aldehyde (TA))
Α, α-trehalose dihydrate (endotoxin-free) (Hayashibara) dried under reduced pressure for 2 hours at 60 ° C (using a water bath manufactured by Yazawa Seisakusho) at 2 mmHg (using a vacuum pump GDH-60 manufactured by Shimadzu Corp.) 19.27 g was dissolved in 200 mL of ion-exchanged water (Kishida Chemical) and charged into a 2 L four-necked flask. Periodic acid dihydrate (Aldrich); 48.75 g was dissolved in ion-exchanged water (Kishida Kagaku); 400 mL and charged into the four-necked flask. 200 mL of ion-exchanged water (Kishida Chemical); 200 mL was charged into the four-necked flask. The four-necked flask was stirred for 22 hours at 21 ° C. (using a low temperature thermostatic chamber CA-1111 manufactured by Tokyo Science Instruments) while stirring under light shielding (using a stirrer PC-30 manufactured by Nonaka Science Instruments). Ion exchange resin (Amberlite IRA96SB: Organo); 1 L was washed 3 times with 500 mL of ion exchange water (Kishida Chemical). The reaction solution was transferred to a 2 L beaker, and an ion exchange resin (Amberlite IRA96SB: Organo) was added while stirring (using an Advantech magnetic stirrer SRS710AA). Ion exchange resin (Amberlite IRA96SB: Organo); 600 mL was added, and it was confirmed that the pH of the aqueous layer (Advantech test paper UNIV) was neutral. The ion exchange resin (Amberlite IRA96SB: Organo) was filtered off (Advantech filter paper No. 2) and washed with ion exchange water (Kishida Chemical) four times with 400 mL. Trehalose aldehyde (TA) was transferred to an eggplant-shaped flask and concentrated under reduced pressure and dried at 35 to 40 ° C. (using a water bath manufactured by Yazawa Seisakusho) with 2 mmHg (vacuum pump GDH-60 manufactured by Shimadzu Corporation). 16.61 g was obtained.

(Adjustment of crosslinker concentration)
The TA was prepared by dissolving in distilled water for injection (Otsuka Pharmaceutical). Diluting cross-linking agents mixed with glutaraldehyde (GA) (glutaraldehyde aqueous solution Wako Pure Chemical Industries), formaldehyde (FA) (formaldehyde liquid Wako Pure Chemical Industries), and glyoxal (GL) (Glyoxal Aldrich) (Otsuka Pharmaceutical). In Examples, when two or more kinds of crosslinking agents are mixed and used, for example, the expression “TA / GA = 10/4” means that 10 wt% TA and 4 wt% GA coexist in the aqueous crosslinking agent solution. Means.

(Main ingredient adjustment-freeze-drying)
This was performed using a vacuum freeze dryer (model; TF5-80MTN, Takara Seisakusho Co., Ltd.). One albumin aqueous solution (25% buminate, Baxter) was poured into one disposable dish (9 cmφ, Nipro). After completely freezing at −42 ° C. for 5 hours, reduced pressure to 0.01 Torr over 1 hour, followed by drying at 25 ° C. for 60 hours and returning to normal pressure, albumin powder (HSA); 13.29 g was obtained.

(Adjustment of main agent-adjustment of HSA aqueous solution)
The HSA was adjusted by adding a predetermined amount of distilled water for injection (Otsuka Pharmaceutical Co., Ltd.). Hereinafter, the adjustment method of 45 weight% HSA (HSA = 45) is shown as a specific example. HSA obtained by freeze-drying in a 50 mL centrifuge tube (polypropylene, IWAKI); 5.40 g and distilled water for injection (Otsuka Pharmaceutical); 6.63 g were weighed in order, and occasionally shaken by hand. It was left to stand in a refrigerator at 0 ° C. for 1 week, and dissolution was confirmed visually.

(Main ingredient adjustment-addition of resorcin or catechin)
Resorcin (R) (Resorcinol Wako Pure Chemical Industries), β-resorcylaldehyde (RA) (2,4-Dihydroxybenzaldehyde Aldrich) or catechin (catechin mixture, green tea derived Wako Pure Chemical Industries), and sodium benzoate (SB) ( After confirming that Wako Pure Chemical Industries) was dissolved in distilled water for injection (Otsuka Pharmaceutical), the above HSA was added and dissolved. For example, “HSA / R / SB = 34/8/3” means that 34, 8, and 3% by weight of HSA, R, and SB are contained in the main agent aqueous solution, respectively. Hereinafter, the adjustment method of (HSA / R / SB = 34/8/3) will be shown as a specific example. In a 50 mL centrifuge tube (polypropylene, IWAKI), R; 0.96 g, SB; 0.36 g was weighed, and distilled water for injection (Otsuka Pharmaceutical); 6.60 g was added and shaken to dissolve completely. Thereafter, 4.08 g of HSA obtained by freeze-drying was added, and the mixture was allowed to stand in a refrigerator at 5 ° C. for 1 week with occasional shaking, and dissolution was confirmed visually.

(Example 1)
Various measurements were performed using the main agent (HSA / R / SB = 34/8/3) and the crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 1.

(Example 2)
Various measurements were performed using a main agent (HSA / R / SB = 34/8/3) and a crosslinking agent (GL / GA = 5 / 3.1). The results are shown in Table 1.

(Example 3)
Various measurements were performed using a main agent (HSA / R / SB = 34/8/3) and a crosslinking agent (TA / GA = 2 / 10.5). The results are shown in Table 1.

Example 4
Various measurements were carried out using a main agent (HSA / catechin / SB = 30/9/9) and a crosslinking agent (TA / GA = 12.5 / 5). The results are shown in Table 1.

(Example 5)
Various measurements were performed using a main agent (HSA / catechin / SB = 30/9/10) and a cross-linking agent (GA = 5). The results are shown in Table 1.

(Example 6)
Various measurements were performed using a main agent (HSA / R = 34/8) and a crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 1.

(Example 7)
Various measurements were performed using a main agent (HSA / R / SB = 34/25/3) and a cross-linking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 2.

(Example 8)
Various measurements were performed using a main agent (HSA / R / SB = 34 / 0.001 / 3) and a crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 2.

Example 9
Various measurements were performed using a main agent (HSA / R / SB = 30/10/21) and a crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 2.

(Example 10)
Various measurements were performed using a main agent (HSA / R / SB = 30/10 / 0.0001) and a crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 2.

(Example 11)
Various measurements were carried out using the main agent (HSA / RA / SB = 30/8/3) and the crosslinking agent (FA / GA = 5.6 / 0.8). The results are shown in Table 2.

(Comparative Example 1)
Various measurements were performed using a main agent (gelatin / R = 37.5 / 12.5) and a crosslinking agent (FA / GA = 18.5 / 2.5). The results are shown in Table 2.

(Comparative Example 2)
Various measurements were performed using a main agent (HSA = 45) and a crosslinking agent (TA / GA = 4/21). The results are shown in Table 2.

  As shown in Tables 1 and 2, by using a main agent and a crosslinking agent having an appropriate concentration and composition, a two-component adhesive having an appropriate main agent viscosity, reaction rate, reaction rate, gel flexibility and gel strength can be obtained. It turns out that it is obtained.

  In the present invention, by using a cross-linking agent combined with an aldehyde in a main agent in which albumin, a polyphenol compound or a polyphenol compound, and an additive are mixed, the main agent has a viscosity that facilitates application to a target site. It was found that an excellent and impossible gel was obtained.

  The two-component adhesive of the present invention does not require a special treatment of preheating as compared with the conventional adhesive, and has an adhesive performance such as an adhesive time and a gelation reaction rate with an appropriate main agent viscosity. This means that a flexible gelled product suitable for living tissue can be obtained. Furthermore, it is possible to provide a material that is particularly effective for optimizing the adhesion time during biological tissue adhesion using a two-component mixing device that enables precise mixing of the main agent and the crosslinking agent. Therefore, it is important to contribute to industrial development.

It is a schematic diagram which shows the difference of the structure of the adhesive agent (gelled material) of this invention, and a GRF adhesive agent (gelled material). It is a schematic diagram which shows the presence state of each component in the main ingredient of the adhesive agent of this invention.

Claims (19)

  A two-component adhesive comprising a main agent and a cross-linking agent, wherein the main agent is mainly composed of albumin and a polyphenol compound and / or a polyphenol compound, and the cross-linking agent is mainly composed of an aldehyde.   The two-component adhesive according to claim 1, wherein the main agent contains an additive.   The two-component adhesive according to claim 2, wherein the additive is a sodium salt of an organic acid.   The two-component adhesive according to claim 2 or 3, wherein the additive is sodium benzoate.   The two-component adhesive according to claim 1, wherein the polyphenol compound is resorcin or β-resorcylaldehyde (2,4-dihydroxybenzaldehyde).   The two-component adhesive according to claim 1, wherein the polyphenol compound is a flavonoid.   The two-component adhesive according to any one of claims 1 to 6, wherein the content of the polyphenol compound and / or polyphenol compound in the main agent is 0.01 to 20% by weight.   The two-component adhesive according to any one of claims 1 to 7, wherein an additive content in the main agent is 0.001 to 20% by weight.   The two-component adhesive according to claim 1, wherein the albumin is human serum albumin.   The two-component adhesive according to any one of claims 1 to 9, wherein the content of albumin in the main ingredient is 5 to 45% by weight.   The two-component adhesive according to claim 1, wherein the aldehyde is an alkylene aldehyde and / or a sugar aldehyde.   The two-component adhesive according to claim 11, wherein the alkylene aldehyde comprises one or more alkylene aldehydes selected from the group consisting of formaldehyde, glyoxal, succinaldehyde, glutaraldehyde, and malealdehyde.   The two-component adhesive according to claim 11 or 12, wherein the content of alkylene aldehyde in the crosslinking agent is 0 to 30% by weight.   The two-component adhesive according to claim 11, wherein the sugar aldehyde is obtained by oxidizing a sugar having a repeating unit of 1 to 1,000.   The two-component adhesive according to claim 11 or 14, wherein the sugar aldehyde is at least one selected from those obtained by oxidizing monosaccharides, disaccharides, trisaccharides and tetrasaccharides.   The two-component adhesive according to claim 11, wherein the sugar aldehyde content in the crosslinking agent is 0 to 30% by weight.   The two-component adhesive according to any one of claims 1 to 16, wherein an aldehyde content in the crosslinking agent is 0.001 to 30% by weight.   The two-component adhesive according to any one of claims 1 to 17, wherein the main agent and the crosslinking agent are mixed at a weight ratio of 100: 1 to 100: 100. The two-component adhesive according to any one of claims 1 to 18, which is an adhesive for hemostasis or biological tissue.

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