WO2006016600A1

WO2006016600A1 - Tartaric acid derivative and high-molecular weight crosslinked matter synthesized by using the derivative

Info

Publication number: WO2006016600A1
Application number: PCT/JP2005/014636
Authority: WO
Inventors: Tetsushi Taguchi; Hisatoshi Kobayashi; Junzo Tanaka; Hirofumi Saito; Hirokatsu Aoki
Original assignee: National Institute For Materials Science; Furuuchi Chemical Corporation
Priority date: 2004-08-10
Filing date: 2005-08-10
Publication date: 2006-02-16
Also published as: JPWO2006016600A1

Abstract

Because of being not natural but artificially synthesized substances, adhesives for biological use and crosslinking agents and condensing agents having been employed in treating medical devices such as cardiac valves exhibit toxicity without being metabolized in vivo. To dissolve them, organic solvents are needed. Thus, it has been required to develop a water-soluble crosslinking agent having a high biocompatibility. A water-soluble tartaric acid derivative obtained by modifying a carboxyl group of tartaric acid and its derivative having a hydroxyl group in molecule with an electron-withdrawing group such as succinimidyl, sulfosuccinimidyl, maleimidyl, phthalimidyl, imidazolyl, nitrophenyl or tresyl or a combination of two or more thereof, and a high-molecular weight crosslinked matter obtained by using this tartaric acid derivative.

Description

Specification

Tartaric acid derivative and crosslinked polymer synthesized by the derivative

Technical field

[0001] The present invention comprises a tartaric acid derivative in which the carboxyl group of tartaric acid or a derivative thereof is modified with an electron-withdrawing group, a polymer crosslinked product synthesized from the tartaric acid derivative, the tartaric acid derivative and a biodegradable polymer. The present invention relates to a two-component biodegradable and absorbable adhesive medical material.

Background art

[0002] So far, medical devices using biological tissue adhesives or biological materials have been used in human-chemically synthesized aldehyde-containing cross-linking agents such as dartal aldehyde or 1-ethyl 3- Uses condensing agents such as (3-dimethylaminopropyl) carposimide! (For example, Patent Documents 1 and 2). The present inventors have developed a low-molecular-weight biological derivative in which at least one carboxyl group of citrate, which is a tricarboxylic acid existing in the citrate circuit, is modified with an electron-withdrawing group (Patent Document 3, Non-Patent Document 3). References 1-3). In addition, a method for producing an intermolecular cross-linked protein using al force nicnic acid disuccinimide is known (Patent Document 4).

Patent Document 1: Japanese Patent Laid-Open No. 9103479

Patent Document 2: JP-A-11 239610

Patent Document 3: Japanese Patent Laid-Open No. 2004-99562

Patent Document 4: JP-A-61-69759

Non-Patent Document 1: Polymer Preprints, Japan 2002, Vol.51, No.14,3728

Non-Patent Document 2: Polymer Preprints, Japan 2003, Vol.52, No.14,4147

Non-Patent Document 3: Polymer Preprints, Japan 2003, Vol.52, No.14,4140

Disclosure of the invention

Problems to be solved by the invention

[0004] Cross-linking agents and condensing agents that have been used so far for medical devices are mainly artificially synthesized non-natural products that are not metabolized in vivo and show toxicity to the living body. thing Has been pointed out. For this reason, the usage and usage are limited when used in the medical field. Furthermore, until now, there has been no biotissue adhesive that has strong tissue adhesion and low residual toxicity of the crosslinking agent or degradation product. In addition, conventional hemostatic agents, vascular embolizers, sealants, or aneurysm sealants are sufficient to easily remove the applied partial force against vascular occlusion, hemostasis, air leak, aneurysm sealing, etc. There was no one with adhesive strength.

[0005] Further, since the alkanedioic acid disuccinimide disclosed in the low-molecular-weight biological derivative developed by the present inventors and Patent Document 4 has low solubility in water, an organic solvent is used for synthesizing a crosslinked product. The use of dimethyl sulfoxide, which is Therefore, development of a water-soluble cross-linking agent that is metabolized and excreted in a living body having high biocompatibility is desired.

Means for solving the problem

In order to solve such problems, in the present invention, a water-soluble tartaric acid derivative obtained by modifying tartaric acid, which is a dicarboxylic acid having a hydroxyl group, with an electron-attracting group, and a biodegradable polymer by using the tartaric acid derivative are used. We have developed a cross-linked polymer synthesized by cross-linking, a two-component biodegradable absorbable adhesive medical material composed of the tartaric acid derivative and a biodegradable polymer.

That is, the present invention crosslinks a biodegradable polymer using a dicarboxylic acid derivative obtained by modifying two carboxyl groups of tartaric acid, which is a dicarboxylic acid, with an electron-attracting group, and the obtained tartaric acid derivative. Is a crosslinked polymer prepared by

[0008] In addition, the present invention provides an organic solvent solution or aqueous solution of a biodegradable polymer, a buffer solution, or a water organic solvent mixed solution as an adhesive component, and two carboxyl groups of tartaric acid that is a dicarboxylic acid are electron-withdrawing. A two-component biodegradable absorbable adhesive medical material characterized in that a tartaric acid derivative modified with a group is used as a curing component.

[0009] The electron-withdrawing group to be used includes one or a combination of two or more of succinimidyl, sulfosuccinimidyl, maleimidyl, phthalimidyl, imidazolyl, nitrophenol, trezyl, and derivatives thereof. It is done.

[0010] Further, the biodegradable polymer used in the crosslinked polymer or biodegradable absorbable adhesive medical material The molecule can be a protein, glycosaminodarlican, chitosan, polyamino acid, polyalcohol, or a combination of two or more thereof.

[0011] The glycosaminodarlicans include chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparan sulfate, heparin, keratan sulfate, or one or a combination of two or more of these derivatives. These glycosaminodaricans do not depend on the molecular weight and the organism from which they are derived.

[0012] Proteins include collagen (depending on several tens of types), atelocollagen (not depending on several tens of types), alkali-treated collagen (not depending on several tens of types), gelatin Selected from the group consisting of macromolecules having amino groups such as keratin, hemoglobin, casein, globulin, fibrinogen, albumin derived from human blood, recombinant albumin, albumin fragment, and chemically modified albumin. A combination of one or more of the qualities can be mentioned.

[0013] In addition, other biodegradable polymers used in the preparation of biodegradable bioabsorbable adhesive medical materials composed of a crosslinked polymer, a water-soluble tartaric acid derivative and a biodegradable polymer. Examples include chitosan (deacetylation degree, regardless of molecular weight), polyamino acid (amino acid type, regardless of molecular weight), and polyalcohol (type, regardless of molecular weight).

[0014] In preparing a bicomponent biodegradable absorbent adhesive medical material composed of a crosslinked polymer, a water-soluble tartaric acid derivative and a biodegradable polymer, the biodegradable polymer Solvents for dissolving include buffers prepared by one or a combination of two or more of hydrochloride, sulfate, nitrate, phosphate, carbonate and borate.

The invention's effect

[0015] The tartaric acid derivative in which the carboxyl group of tartaric acid or its derivative is modified with an electron-withdrawing group is more water-soluble than the low molecular weight biological derivatives and alkanedioic acid disuccinimide previously developed by the present inventors. Biodegradable under the condition of aqueous solution using a buffer solution when obtaining a biodegradable bioadhesive adhesive medical material composed of a crosslinked polymer and the tartaric acid derivative and a biodegradable polymer molecule. The reaction proceeds uniformly with the aqueous polymer solution and the reaction time is shortened.

BEST MODE FOR CARRYING OUT THE INVENTION [0016] The low molecule used as a starting material of the water-soluble tartaric acid derivative of the present invention is tartaric acid or a derivative thereof, for example, a hydroxyl group (-OH) in tartaric acid is converted to a sulfate group (-OSO Na) or a carboxylic acid.

Three

It is chemically modified to a boxymethyl group (-〇CH C000Na). The water of the present invention

2

Soluble tartaric acid derivatives have two carboxyl groups in tartaric acid or its derivatives, e.g., electron-withdrawing groups such as succinimidyl, sulfosuccinimidyl, maleimidyl, phthalimidyl, imidazolyl, nitrophenol, toresyl or their derivatives. A synthetic reaction is carried out with one or a combination of two or more, and an active ester is introduced.

[0017] The water-soluble tartaric acid derivative of the present invention can be obtained by the following synthesis reaction. Tartaric acid or a derivative thereof is added to about 0.001 to 10% by weight, more preferably about 1 to 3% by weight with respect to 100% by weight of the organic solvent, and a condensing agent such as 1-ethyl 3- (3- Dimethylaminopropyl) carbodiimide (EDC) or dicyclohexylcarbodiimide (DCC) is added in an amount of about 0.001 to 20% by weight, more preferably about 5 to 15% by weight, based on 100% by weight of the solvent. In the presence thereof, a molecule that becomes an electron-withdrawing group, for example, N-hydroxysuccinimide is added in an amount of about 0.001 to about LO weight%, more preferably about 3 to 8 weight% with respect to 100 weight% of the solvent. In addition, reaction temperature 0 to: L00 ° C., more preferably 10 to 30 ° C., reaction time 1 to 48 hours, more preferably 1 to 3 hours. The acid derivative and reaction by-product DCC urea precipitate.

[0018] Thereafter, the tartaric acid derivative is isolated by filtering DCC urea as a reaction by-product through a glass filter. After the filtrate is distilled off under reduced pressure, the resulting residue is purified by washing with an organic solvent such as n-hexane and filtering. Exceeding the above reaction conditions is inappropriate because the carboxyl group in tartaric acid cannot be modified with N-hydroxysuccinimide, which is an electron-absorbing I-group. By this reaction, a water-soluble tartaric acid derivative represented by the following structural formula is obtained. This water-soluble tartaric acid derivative is useful as a crosslinking agent, a curing agent for medical adhesives, and a fixing agent such as a pig valve (Carpentier-Edowards valve: CE valve).

[0019] [Chemical 1]

[0020] The cross-linking reaction between the water-soluble tartaric acid derivative of the present invention and the biodegradable polymer has a concentration of the tartaric acid derivative of about 0.1 to 50% by weight of the biodegradable polymer in the solvent. The reaction is carried out at about 01% to 50% by weight, preferably about 10 to 50 ° C. Exceeding the above reaction conditions is unsuitable because the reaction rate becomes slow, the crosslinking density of the resulting crosslinked product becomes low, and a crosslinked product may not be obtained. It is preferable to dissolve the tartaric acid derivative in the above-mentioned solvent and mix them as a solution having an appropriate concentration so that the concentration is within the above-mentioned concentration range in order to cause a uniform reaction.

[0021] As the solvent, a dimethyl sulfoxide solution or a buffer solution having a combination force of one or more of hydrochloride, sulfate, nitrate, phosphate, carbonate and borate, or the buffer solution-dimethyl sulfoxide A mixed solution or the like can be used.

[0022] By this reaction, a crosslinked product having a structure as shown in FIG. 1 is obtained. This crosslinked product is a hyde mouth gel having a water content of about 80 to 99% by weight. When the moisture content is lowered, the strength is increased and the crosslink density is increased, so that the usefulness to an adhesive is improved. This crosslinked product can be used for medical adhesives, hemostatic agents, vascular embolization agents, aneurysm sealing agents, and the like.

[0023] When the polymer crosslinked product produced as described above is applied to any one of a bioadhesive, a hemostatic agent, a vascular embolization agent, and an aneurysm sealant, the crosslinking reaction is directly performed on the affected part. . In addition, once used after a crosslinking reaction, it is suitably used as an anti-adhesion agent, a scaffold material for tissue regeneration, and a drug carrier.

[0024] The cross-linking reaction between the water-soluble tartaric acid derivative of the present invention and the biodegradable polymer used in the biodegradable and absorbable adhesive medical material is performed by the concentration of the biodegradable polymer in the solvent of 0.1 to 50% by weight. The concentration of the tartaric acid derivative is about 0.01% to 50% by weight, preferably 10%. React at about 50 ° C. If the reaction conditions are not met, the adhesion time becomes longer and the adhesion strength also becomes weaker. It is preferable that the tartaric acid derivative is also dissolved in the solvent and mixed as a solution having an appropriate concentration so that the concentration is within the above-mentioned concentration range in order to allow the two components to react uniformly.

[0025] Examples of the solvent include a dimethyl sulfoxide solution or a buffer solution having a combination force of one or more of hydrochloride, sulfate, nitrate, phosphate, carbonate and borate, or the buffer solution-dimethylsulfoxide. A mixed solution or the like can be used. As shown in FIG. 2, the structure of the interface between the biological tissue and the adhesive obtained by the above reaction is such that the tartaric acid derivative undergoes a crosslinking reaction with the biopolymer in the biological tissue and the biodegradable polymer in the adhesive. Adhere by.

Example 1

Hereinafter, the present invention will be described in detail with reference to examples.

Dicyclohexylcarbodiimide (DCC) is added in an amount of 14% by weight with respect to 100% by weight of a solvent in which 5% by weight of tartaric acid is added to 100% by weight of a tetrahydrofuran (THF) solvent. 8% by weight of succinimide was added to 100% by weight of the solvent, and the mixture was stirred for 1 hour, and then stirred at room temperature for 2 hours. As a result, a mixed solution of DCC urea and a tartaric acid derivative was obtained.

[0027] Subsequently, the precipitated DCC urea was removed with a glass filtration filter, and THF as a solvent in the reaction system was distilled off under reduced pressure. The resulting residue was purified by washing with n-xane and 2-propanol and filtered to synthesize tartaric acid derivative (TAD) modified with two carboxyl group N-hydroxysuccinimides of tartaric acid. It was confirmed by 1H—NMR that the two forces of tartaric acid were completely replaced by N-hydroxysuccinimide. The synthesized TAD could be dissolved in water at a concentration of 0.150% by weight.

Example 2

<0028> <Synthesis of cross-linked collagen>

Using TAD synthesized in Example 1, a polymer bridge of alkali-treated collagen (A ¾1) A bridge body was synthesized. 0. Dissolve TAD100 1 in 1M phosphate buffer solution (pH 7.0). 1 concentration 2.5 wZv% phosphate buffer solution (0.1 Μ, ρ Η 7.0) was added in 4 ΟΟ / ζ 1 and stirred. The final concentration of TAD was added to 20, 50, 100, and 150 mM. Thereafter, the mixture was allowed to stand at 37 ° C for 30 minutes, and the presence or absence of collagen cross-linked body formation was confirmed. The results are shown in Table 1. From the results in Table 1, it was confirmed that a 0.1 M phosphate buffer solution, that is, the formation of a cross-linked collagen under aqueous solution conditions. When the moisture content is lowered, the strength is increased and the crosslink density is increased, so that the usefulness to an adhesive and the like is improved.

[0029] [Table 1]

Example 3

<0030> <Synthesis of albumin cross-linked product>

Using the TAD synthesized in Example 1, a crosslinked polymer of human serum albumin (HSA) was synthesized. 0.1. TAD 100 1 was dissolved in 1M phosphate buffer solution (pH 7.0), and added to phosphate buffer solution (0.1M, pH 7.0) 4OO 冲 with HSA concentration of 45w Zv% and stirred. The final concentration of TAD was 100, 150, 200 mM. Thereafter, the mixture was allowed to stand at 37 ° C for 30 minutes, and the presence or absence of HSA crosslinked product formation was confirmed. The results are shown in Table 2. From the results in Table 2, it was confirmed that 0.1M phosphate buffer solution, that is, gelation of biopolymers under aqueous solution conditions.

[0031] [Table 2] Tartaric acid derivative concentration. (MM) Moisture content of crosslinked albumin (weight ⁰ / o)

100 76

150 76

200 78 Example 4

[0032] Preparation of biological tissue adhesive>

A biological tissue adhesive was prepared as follows. Albumin derived from human serum (A1653 manufactured by Sigma Aldrich Japan Co., Ltd.) was dissolved in 0.1 M sodium phosphate buffer (pH 7.0) to a concentration of 45% by weight. To this albumin solution 4001, 100 M / l of a 0.1 M phosphate buffer solution (pH 7.0) of TAD as a curing component was added and stirred for several seconds at 25 ° C. The TAD concentration with respect to the final volume (500 ul) was adjusted to lOOmM, 150mM, 200mM, and 250mM.

[0033] Measurement of adhesive strength of biological tissue adhesives>

Collagen casing (made by Nitta Gelatin Co., Ltd., composition: collagen 44%, cellulose 18%, glycerin 15%, vegetable oil 3%, carboxymethylcellulose 2%) as an adherend for measuring the adhesive strength to living tissue Used to measure the adhesive strength. Apply 50 folds of the mixed solution before hardening to a 10 mm x 10 mm area of a collagen casing (width 10 mm x length 25 mm), and place an equal-sized collagen casing on the bonding surface. Superimposed. Further, a 50 g weight was placed on the adhesive surface and allowed to stand at 37 ° C for 1 hour. The adhesive strength was measured with a tensile tester (TA-XT2i manufactured by Eihiro Seiki Co., Ltd.). Measurement conditions were 25 ° C and a measurement speed of 2 mm / s. The results are shown in Table 3. From the results in Table 3, it was confirmed that the adhesive strength became stronger depending on the TAD concentration.

[0034] [Table 3]

TAD concentration (mM) Adhesive strength (gZcm ² )

1 00 1 77. 6

1 50 356. 2

200 676. 8

250 660. 7 [0035] In addition, a fibrin-based adhesive (product name: Bolheel, manufactured by Fujisawa Pharmaceutical Co., Ltd.) and a cyanacrylate-based adhesive (product name: Dermabond, manufactured by ETHICON), which are tissue adhesives for living bodies, are commercially available. In the same manner, the adhesive strength was measured. The results are shown in Table 4. From the results in Table 4, it was confirmed that TAD adhesives are 10 times higher in adhesive strength than fibrin adhesives, although they are not as good as cyanoacrylate adhesives.

[0036] [Table 4]

Industrial applicability

[0037] The water-soluble tartaric acid derivative of the present invention is useful as a cross-linking agent for medical materials such as bioadhesives, hemostatic agents, vascular embolic agents, and aneurysm sealants. In addition, once used after cross-linking reaction with a biodegradable polymer, it can also be used as an anti-adhesive agent, a scaffold material for tissue regeneration, and a drug carrier.

Brief Description of Drawings

FIG. 1 is a schematic view showing the structure of a crosslinked product of the present invention.

FIG. 2 is a schematic diagram showing a structure of an interface between a biological tissue and an adhesive when the biological tissue adhesive of the present invention is used.

Claims

The scope of the claims

[1] One or two of succinimidyl, sulfosuccinimidyl, maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, tresyl or their derivatives, which are two electron-withdrawing groups of tartaric acid or its derivatives A water-soluble tartaric acid derivative characterized by being modified by the above combination.

[2] A cross-linked water-soluble polymer, characterized in that the biodegradable polymer power is crosslinked using the water-soluble tartaric acid derivative according to claim 1.

[3] The water-soluble tartaric acid derivative according to claim 1 as a curing component, and a dimethyl sulfoxide solution or a combination of one or more of hydrochloride, sulfate, nitrate, phosphate, carbonate, borate A two-component biodegradable absorbable adhesive medical material characterized by comprising as an adhesive component a buffer solution comprising the above, a mixed solution of the buffer solution and dimethyl sulfoxide, or a biodegradable polymer dissolved in a powerful solvent.

[4] Soft tissue and soft tissue, soft tissue and hard tissue, or biological tissue that bonds hard tissue and hard tissue, characterized in that the two-component biodegradable absorbable adhesive medical material force according to claim 3 .

[5] A hemostatic agent, a vascular embolization agent, a sealant, or an aneurysm sealant, characterized in that the two-component biodegradable and absorbable adhesive medical material according to claim 3 is used.

[6] Using the water-soluble tartaric acid derivative according to claim 1, from a dimethyl sulfoxide solution or one or a combination of two or more of hydrochloride, sulfate, nitrate, phosphate, carbonate and borate. A method for synthesizing a polymer crosslinked product comprising cross-linking a biodegradable polymer in a buffer solution or a mixed solution of dimethyl sulfoxide.