JP2007252699A - Gel composition, material for transplantation using the same, and their manufacturing method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition which is used for a medical article such as an artificial blood vessel, a patch for repairing a cardiovascular, which never depends on human/animal-originated raw material to be safety, which is antithrombogenic, where leakage of blood is avoided both during and after a surgery, and which is decomposed gradually to be discharged from the kidneys without causing abnormality in the internal organs after a neoblastic is grown to the degree of avoiding leakage from the whole blood vessel when one month or longer than this passes after the surgery, and to provide material for transplantation using the composition and an article for transplantation. <P>SOLUTION: The gel composition is obtained by amidating free amino residue which is not connected with gelatinization in hydrogel obtained by reaction of chain polysaccharide derivative which is obtained by introducing a succinimide residue into a side chain, and polyamine or its derivative. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a gel composition, a transplant material using the same, and a method for producing them. More specifically, the present invention relates to a medical coating material or sealing material composition that does not use human or animal-derived raw materials such as gelatin, collagen, albumin, etc., and a transplant coated or sealed using the same. The present invention relates to a material for transplantation and an article for transplantation, in particular, a transplantation material excellent in blood compatibility and an article for transplantation, and a method for producing them.

  Artificial joints and dental implants made of hard materials have been conventionally known as artificial transplant articles that are transplanted for the purpose of being maintained for a certain period of time in a living body. Also, patches for cardiovascular repair, artificial pericardium, artificial dura mater, etc. are widely used. As a base material for such an implantable article, woven fabric, knitted fabric or porous polytetrafluoroethylene made of polyester fiber typified by polyethylene terephthalate is used because of its in vivo degradation resistance and harmlessness to surrounding tissues. Used exclusively.

  When the former polyester fiber cloth is embedded as an artificial blood vessel or patch that comes into contact with blood, a new tissue derived from the living body gradually grows in the void to close the pores and build an inner surface that can coexist stably with blood for a long period of time. To do. This ability is expressed by the magnitude of the cloth's water leak, called porosity. That is, it is known that the coarser the blood leaks immediately after transplantation, the better the new tissue can be formed. On the other hand, however, it is also essential to prevent blood leakage during surgery. Currently, products containing compositions that are coated with human or animal-derived proteins such as gelatin, collagen, and albumin and cross-linked as necessary. Is commercially available. As the cross-linking agent, dialdehyde represented by glutaraldehyde, formaldehyde, carbodiimide, or isocyanate is used.

  Examples of these transplant materials include U.S. Pat. No. 4,747,848 (Patent Document 1), U.S. Pat. No. 4,902,290 (Patent Document 2), and JP-A-61-135651 (Patent Document 3). It is done.

However, human-derived raw materials such as albumin cannot deny the risk of infectious diseases such as AIDS and viral hepatitis. Regarding the use of animal-derived raw materials, the risk of infectious diseases such as bovine spongiform encephalopathy cannot be ruled out, so the use of alternative substances is recommended for use. It is costly and includes risky elements.
In addition, each of the above patent documents discloses a transplant material using gelatin. However, gelatin has been pointed out as a cause of shock in addition to many problems of late-onset fever. There is a need for replacement with objects.

Furthermore, since the covering material applied to the artificial blood vessel or cardiovascular repair material is premised on contact with the circulating blood, it is different from a material used in the abdominal cavity (for example, a medical sheet for hernia repair). There are important quality requirements. That is,
(1) Do not decompose at least for a period of about one month or more until the neointima is stabilized,
(2) Due to the rapid dissolution and biodegradation, the water absorbs and swells rapidly, the strength decreases, and it has the property of peeling off into the circulating blood with arterial pulsation and not flowing away,
(3) Dissolved decomposition products diffuse without adversely affecting the surrounding tissue, cause abnormalities in organs, and are discharged from the kidneys; and
(4) If a large amount of blood clot is generated at the blood contact surface after transplantation, it may peel off and flow to the periphery, causing organ necrosis, or in the case of thin blood vessels, it may cause a serious problem of reducing the patency performance Therefore, it is essential to have anti-thrombogenicity that does not inhibit neointimal growth and at the same time does not rapidly form thrombus.

For example, Japanese Patent Publication No. 2003-508158 (Patent Document 4) describes that dextran is cross-linked using formaldehyde and urea. Since all degradation products are low-molecular, it is easy to excrete and is characterized by fast biodegradation.
The optimal molecular weight of dextran is 40,000. The substance produced as a result of crosslinking with such a low molecular weight compound is a low molecular weight compound, but it decomposes quickly and ensures a decomposition period of about one month necessary for waiting for complete healing so as not to cause plasmaoma in the body. In order to do this, the cross-linking must be strengthened. For that purpose, it is necessary to increase the contents of urea and formalin, and the physical properties become hard and brittle. In order to avoid this and maintain the flexible and flexible physical properties that are not necessary as an artificial blood vessel, there is a contradiction that the decomposition speed increases as a result. The examples clearly state that dogs have been absorbed within one month under all conditions.

  In the above-mentioned Japanese Patent Laid-Open No. 61-135651 (Patent Document 3), a technique for increasing the decomposition speed is disclosed. When this method is used, the gel dissolves or dissolves or dissolves in 1 to 2 weeks. In Japan, however, it has been reported to academic societies from the early stage of occurrence of plasmaoma. There are few reports of subacute complications such as plasma tumors caused by artificial blood vessels in the United States and Europe, but this is because subacute complications such as plasma tumors occur in artificial blood vessels with a rapid degradation rate in Europe and the United States where hospitalization is short. However, it is thought that it is difficult for doctors to grasp, and basically it is considered that a decomposition period of about one month is necessary.

  As described above, the above-described problems cannot be solved by the techniques that have been publicly known. In other words, various synthetic materials that satisfy physical properties suitable for surgery have been proposed, but they are comprehensively evaluated based on their biological safety, biodegradability, antithrombotic properties, and curative properties (properties that make neointima). There are no clinically usable levels. As a result, despite the fact that various human viral infections and human communicable communicable diseases have become a problem today, the reality is that they have not been able to escape from human and animal-derived raw materials.

U.S. Pat. No. 4,747,848 US Pat. No. 4,902,290 JP 61-135651 A Special table 2003-508158 gazette

  Therefore, the object of the present invention is to apply to blood contact medical articles such as artificial blood vessels and patches for cardiovascular repair, and has high safety and good antithrombogenicity that do not depend on human or animal-derived materials. Moreover, blood leakage does not occur during the operation or in the recovery period after the operation, and after about 1 month or more after the operation, the new tissue grows so that it does not leak from the whole blood vessel and gradually decomposes, An object of the present invention is to provide a composition excreted from the kidney without causing any abnormality in the organ, and a transplant material and transplant article using the composition.

  The present inventors have proposed a medical composition by means of reacting a first liquid containing a chain polysaccharide derivative and a second liquid containing a polyamine as a hemostatic material or various medical sheets in Japanese Patent Application Laid-Open No. 2004-359895. . This composition has the property of promoting coagulation with respect to blood, and is characterized by high hemostatic performance in addition to rapid gelation performance. However, as a result of continuing further research and conducting extensive experiments, it was confirmed that by applying acylation after the reaction in the composition, high resistance required for artificial blood vessels with relatively small diameters, parts close to the heart, carotid arteries, etc. The present inventors have found that the property of satisfying thrombosis and functioning as a barrier for blood and body fluid for a certain period can be realized. That is, a cross-linked product synthesized with a chain polysaccharide derivative in which a succinimide residue is introduced into a side chain within a predetermined range and a polyallylamine, or a derivative acylated so as to leave at least a part of its amino group is reacted. It was found that a gel in which the remaining amino group was converted to amide by acylation later solved all the above-mentioned problems, and the present invention was completed.

（式中、Ｙは置換されていてもよいアルキレンまたはカルバモイル基を表す。）で表されるスクシンイミド含有基で置換してなるものである前記１または２に記載のゲル組成物。
４．スクシンイミド残基を側鎖に導入した鎖状多糖誘導体が主要構造として下記式（II）：

（式中、Ｒ¹は水素原子、金属原子、炭素数１ないし３０の置換されていてもよい脂肪族炭化水素基もしくは芳香族炭化水素基または下記式（Ｉ）：

（式中、Ｙは置換されていてもよいアルキレンまたはカルバモイル基を表す。）で表されるスクシンイミド含有基もしくは下記式（III）：

（式中、Ｙは上に定義した通りであり、Ｒ²は水素原子、金属原子、炭素数１ないし３０の置換されていてもよい脂肪族炭化水素基もしくは芳香族炭化水素基を表す。）で表される基であり、Ｘは、−ＯＲ¹、−ＮＨＲ¹（式中、Ｒ¹は上記定義と同じ。）またはカルバモイル基であり、分子中のＲ¹はそれぞれ同一でも異なってもよいが、その少なくとも１個は前記式（Ｉ）のスクシンイミド含有基である。）を繰り返し単位として含む糖鎖である前記３に記載のゲル組成物。
５．スクシンイミド残基を側鎖に導入した鎖状多糖誘導体の主要構造が下記式（IV）：

（式中、Ｒ¹及びＸは前記定義の通りであり、ｎは３以上の整数である。）で表される直鎖状誘導体である前記４に記載のゲル組成物。
６．鎖状多糖誘導体における前記Ｙが炭素数１ないし３のアルキレンまたは下記式（Ｖ）；

（式中、Ｒ³は水素原子、または炭素数１ないし３０の置換されていてもよい脂肪族炭化水素基もしくは芳香族炭化水素基を表す。）で表される基である前記３〜５のいずれかに記載のゲル組成物。
７．ポリアミンが側鎖に少なくとも１モル％以上の遊離のアミノ基を有するポリアリルアミンである前記１〜６のいずれかに記載のゲル組成物。
８．ポリアミンが分子量５００〜１００００である前記１〜７のいずれかに記載のゲル組成物。
９．多糖誘導体が分子量２５万以下である前記１〜８のいずれかに記載のゲル組成物。
１０．前記１〜９のいずれかに記載のゲル組成物を基材中に有するゲル組成物−基材複合材料。
１１．基材が管状、シート状もしくは多孔質またはこれらの組み合わせである前記１０に記載のゲル組成物−基材複合材料。
１２．基材がポリエステル樹脂、及びフッ素樹脂から選択される前記１０に記載のゲル組成物−基材複合材料。
１３．前記１〜９のいずれかに記載のゲル組成物または前記１０〜１２のいずれかに記載のゲル組成物−基材複合材料を含む循環血液もしくは体内の粘膜に直接接触する移植材料。
１４．前記１３に記載の移植材料を含む人工血管。
１５．前記１３に記載の移植材料を含む血液適合性の臓器修復用パッチ、人工心膜、人工硬膜、癒着防止膜または人工皮膚。
１６．前記１５に記載の臓器修復用パッチである心臓血管修復用パッチ。
１７．スクシンイミド残基を側鎖に導入した鎖状多糖誘導体を含む第１液またはポリアリルアミンもしくはその誘導体を含む第２液の一方を塗布して、次いで他方の液を塗布または噴霧または浸漬することにより基材をゲルで被覆した後、残存アミノ基をアシル化剤と反応させる工程を含むゲル組成物−基材複合材料の製造方法。
１８．アシル化剤がアセチル化剤である前記１７に記載のゲル組成物−基材複合材料の製造方法。 That is, the present invention provides the following gel composition, transplantation material and transplantation article using the same, and methods for producing them.
1. A gel composition characterized by amidating a free amino residue that was not involved in gelation in a hydrogel obtained by reacting a chain polysaccharide derivative having a succinimide residue introduced into the side chain with polyamine or a derivative thereof .
2. 2. The gel composition according to 1 above, wherein the chain polysaccharide derivative having a succinimide residue introduced in the side chain contains one succinimide per at least 20 sugar units.
3. A chain polysaccharide derivative in which a succinimide residue is introduced into the side chain, the hydrogen atom of the hydroxyl group of the sugar is represented by the following formula (I):

(1) The gel composition according to the above (1) or (2), wherein the gel composition is substituted with a succinimide-containing group represented by the formula (wherein Y represents an optionally substituted alkylene or carbamoyl group).
4). A chain polysaccharide derivative having a succinimide residue introduced in the side chain is represented by the following formula (II):

(In the formula, R ¹ is a hydrogen atom, a metal atom, an optionally substituted aliphatic hydrocarbon group or aromatic hydrocarbon group having 1 to 30 carbon atoms, or the following formula (I):

(Wherein Y represents an optionally substituted alkylene or carbamoyl group) or a succinimide-containing group represented by the following formula (III):

(In the formula, Y is as defined above, and R ² represents a hydrogen atom, a metal atom, an optionally substituted aliphatic hydrocarbon group or an aromatic hydrocarbon group having 1 to 30 carbon atoms.) X is —OR ¹ , —NHR ¹ (wherein R ¹ is the same as defined above) or a carbamoyl group, and R ^{1 in} the molecule may be the same or different. However, at least one of them is a succinimide-containing group of the above formula (I). 4. The gel composition according to 3 above, which is a sugar chain containing a repeating unit as a repeating unit.
5). The main structure of a chain polysaccharide derivative having a succinimide residue introduced in the side chain is represented by the following formula (IV):

(Wherein R ¹ and X are as defined above, and n is an integer of 3 or more).
6). Y in the chain polysaccharide derivative is alkylene having 1 to 3 carbon atoms or the following formula (V);

(Wherein R ³ represents a hydrogen atom, or an optionally substituted aliphatic hydrocarbon group or aromatic hydrocarbon group having 1 to 30 carbon atoms). The gel composition according to any one of the above.
7). 7. The gel composition according to any one of 1 to 6 above, wherein the polyamine is a polyallylamine having at least 1 mol% or more free amino groups in the side chain.
8). 8. The gel composition according to any one of 1 to 7 above, wherein the polyamine has a molecular weight of 500 to 10,000.
9. 9. The gel composition according to any one of 1 to 8 above, wherein the polysaccharide derivative has a molecular weight of 250,000 or less.
10. The gel composition-base material composite material which has the gel composition in any one of said 1-9 in a base material.
11. 11. The gel composition-base material composite material according to 10, wherein the base material is tubular, sheet-like, porous, or a combination thereof.
12 11. The gel composition-base material composite material according to 10, wherein the base material is selected from a polyester resin and a fluororesin.
13. 10. A transplant material that directly contacts circulating blood or a mucous membrane in the body, comprising the gel composition according to any one of 1 to 9 or the gel composition-substrate composite material according to any one of 10 to 12.
14 14. An artificial blood vessel comprising the transplant material according to 13 above.
15. A blood compatible organ repair patch, artificial pericardium, artificial dura mater, anti-adhesion membrane or artificial skin comprising the transplant material according to the above 13.
16. A patch for cardiovascular repair, which is the patch for organ repair described in 15 above.
17. Applying one of the first liquid containing a chain polysaccharide derivative having a succinimide residue introduced into the side chain or the second liquid containing polyallylamine or a derivative thereof, and then applying or spraying or dipping the other liquid. A method for producing a gel composition-base material composite material comprising a step of reacting a residual amino group with an acylating agent after coating a material with a gel.
18. 18. The method for producing a gel composition-base material composite material according to 17 above, wherein the acylating agent is an acetylating agent.

  The gel composition of the present invention is not derived from humans or animals, has good antithrombotic properties, and blood leakage does not occur during or after the recovery period, and more than one month after the operation. The new tissue grows so that it does not leak from the whole blood vessel and then gradually decomposes and is excreted from the kidney without causing any abnormalities in the organ. For this reason, it is particularly useful as a material for transplantation, a material for transplantation, in particular, a material for covering or sealing an artificial blood vessel in contact with blood flow.

Hereinafter, the present invention will be described in detail.
(A) Chain polysaccharide derivative The chain polysaccharide derivative component in which the succinimide residue used in the present invention is introduced into the side chain is a linear polysaccharide containing a single sugar or a plurality of sugars as a repeating unit. A succinimide residue is introduced into the induced side chain. The sugar is not limited as long as it forms a chain polysaccharide, and includes D-glucose, D-mannose, D-fructose, D-galactose, D-xylose, D-arabinose or derivatives thereof, and homopolysaccharide Alternatively, either a heteropolysaccharide may be used. In addition, the linkage between sugars includes α (1 → 4), α (1 → 6), β (1 → 4) and the like, and therefore may include a branched chain. A molecule in which the length direction of the molecule can be defined.

を繰り返し単位として含む糖鎖であり、式中、Ｒ¹が以下の４群のいずれかの基である。 The chain polysaccharide derivative component is preferably the following compound:

In the formula, R ¹ is any ^one of the following 4 groups.

（式中、Ｙは置換されていてもよいアルキレンまたはカルバモイル基を表す。）で表されるスクシンイミド含有基；及び
４）下記式（III）：

（式中、Ｙは上に定義した通りであり、Ｒ²は水素原子、金属原子、炭素数１ないし３０の置換されていてもよい脂肪族炭化水素基もしくは芳香族炭化水素基を表す。）で表される基。 1) a hydrogen atom;
2) an optionally substituted aliphatic hydrocarbon and / or aromatic hydrocarbon having 1 to 30 carbon atoms;
3) The following formula (I):

(Wherein Y represents an optionally substituted alkylene or carbamoyl group); and 4) the following formula (III):

(In the formula, Y is as defined above, and R ² represents a hydrogen atom, a metal atom, an optionally substituted aliphatic hydrocarbon group or an aromatic hydrocarbon group having 1 to 30 carbon atoms.) A group represented by

The first group is the original hydroxyl group of sugar.
The second group is a group obtained by etherifying or esterifying the original hydroxyl group of sugar and includes various useful modifications. The aliphatic hydrocarbon group or the aromatic hydrocarbon group is not particularly limited, and the aliphatic hydrocarbon group may be linear, branched, or cyclic, and may contain any unsaturated group, but includes 3) group succinimide A chain length of 1 to 30 carbons is preferred so as not to interfere with the reaction of the group. These aliphatic hydrocarbon groups or aromatic hydrocarbon groups may have any substituent as long as they do not cause harmful reactions or decomposition in vivo, and if chemically possible, carbon atoms in the chain Including those replaced by. Similarly, the aromatic may be a heterocycle. Examples of such substituents include a hydroxyl group, an oxy group, an amino group, a carbonyl group, a carboxyl group, a mercapto group, and a thioether group.
Group 3) is an indispensable group for the gel formation of the present invention, and therefore the chain polysaccharide derivative component must contain at least one succinimide-containing group as R ¹ . At least one succinimide per 20 saccharide units, more preferably at least 1 per saccharide unit.

（式中、Ｒ³は前記Ｒ²と同様に水素原子、金属原子または炭素数１ないし３０の置換されていてもよい脂肪族炭化水素基もしくは芳香族炭化水素基を表す。）で表される基である。 Group 4) is its precursor. Therefore, Y in the third group and the fourth group is a group derived from a compound having reactivity with a hydroxyl group of a saccharide and does not hinder introduction of a succinimide group. Such groups include alkylene or carbamoyl groups. The alkylene is preferably alkylene having 1 to 30 carbons, more preferably 1 to 20 carbons, still more preferably 1 to 10 carbons, and most preferably 1 to 5 carbons. The carbamoyl group includes, for example, the following formula (V)

(In the formula, R ³ represents a hydrogen atom, a metal atom, or an optionally substituted aliphatic hydrocarbon group or aromatic hydrocarbon group having 1 to 30 carbon atoms in the same manner as R ² described above). It is a group.

（式中、ｐは１〜５の整数である。）、

で表される基が挙げられる。式中、Ｒ³は前記定義と同じであるが、−ＮＨＣＨ（Ｒ³）ＣＯＯ−が、いわゆる必須アミノ酸及びその慣用される誘導体に由来する基であることが望ましい。なお、ここでいう必須アミノ酸としては、グリシン、アラニン、アルギニン、アスパラギン酸、システイン、シスチン、グルタミン酸、グルタミン、ヒスチジン、リシン、ロイシン、イソロイシン、メチオニン、フェニルアラニン、プロリン、バリン、チロシン、スレオニン、トリプトファン、セリン等が挙げられ、慣用されるその誘導体にはヒドロキシプロリン、ヒドロキシリシン等が挙げられる。
また、式（Ｉ）中、Ｘは、−ＯＲ¹、−ＮＨＲ¹、またはカルバモイル基であり、式中、Ｒ¹及びカルバモイル基は上記定義と同様である。 Accordingly, examples of preferred succinimide-containing R ¹ groups include:

(Wherein p is an integer of 1 to 5),

The group represented by these is mentioned. In the formula, R ³ has the same definition as above, but —NHCH (R ³ ) COO— is preferably a group derived from a so-called essential amino acid and a commonly used derivative thereof. The essential amino acids here include glycine, alanine, arginine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, histidine, lysine, leucine, isoleucine, methionine, phenylalanine, proline, valine, tyrosine, threonine, tryptophan, serine. The derivatives thereof commonly used include hydroxyproline, hydroxylysine and the like.
In the formula (I), X is —OR ¹ , —NHR ¹ , or a carbamoyl group, and in the formula, R ¹ and the carbamoyl group are the same as defined above.

式中、Ｒ¹及びＸは前記定義の通りであり、ｎは３以上の整数である。Ｘの種類に応じて、セルロース誘導体（−ＯＲ¹）、キトサン誘導体（−ＮＨＲ¹）またはキチン誘導体（カルバモイル基）である。 The chain polysaccharide derivative is preferably a saccharide of the following formula (IV).

In the formula, R ¹ and X are as defined above, and n is an integer of 3 or more. Depending on the type of X, it is a cellulose derivative (—OR ¹ ), a chitosan derivative (—NHR ¹ ) or a chitin derivative (carbamoyl group).

In the case of using a cellulose derivative, in the case of a cellulose derivative formed after the gel is decomposed, is it excreted sufficiently because the main route is dispersion from the transplanted site and filtration from the urine in the polymer state? The influence of molecular weight is great on whether or not.
From the results of the experiment, the molecular weight is 250,000 daltons or less, preferably 125,000 daltons or less. If it was in this range, it was excreted from the kidney, and no abnormalities were caused in the major organs as a result of implantation for 6 months.

(B) Polyamine In this invention, a polyamine is used as a compound which bridge | crosslinks the said linear polysaccharide. As an example of a preferable compound, polyallylamine is used. Preferably, polyallylamine or an acylated product thereof having a molecular weight of 500 or more and 10,000 or less, more preferably a molecular weight of about 1000 to 8000, and still more preferably a molecular weight of about 2000 to 5000 is used. If the molecular weight is less than 500, the resulting gel lacks flexibility. Although a flexible physical property can be obtained by using a polymer having a high molecular weight as the polyamine, an excessive molecular weight is disadvantageous for excretion from the body. In addition, there is a problem that the viscosity is increased and the handleability is lowered.

In the present invention, it is indispensable to secure the antithrombogenicity by modifying the —NH ₂ group in the gel that was not involved in the reaction with respect to the gel after completion of the reaction with an acylating agent. These may be amidated with an acylating agent in advance. That is, polyallylamine in which a part of the amino residue is acylated can also be used. Suitable molecular weights and the like are the same as described above.

The proportion to be acylated in advance must be determined in consideration of the necessary physical and chemical properties depending on the degree of substitution of the succinimidyl group per unit of the first compound succinimidated chain hydrocarbon. . For example, in an artificial blood vessel or a cardiovascular surgical patch, it is necessary to endure a surgical operation such as a force for supporting a suture thread, cutting with scissors, picking with tweezers, and bending. In order to increase the crosslinking reaction rate and to increase the crosslinking density of the gel, it is possible to leave a residue that is not acetylated in an amount of 1 mol% or more, preferably 10 mol% or more, and most preferably 20 mol% or more. is necessary.
In order to adjust the hydrophilicity, polyvinylamine may be mixed.

(C) Gelation reaction One of the first liquid containing a chain polysaccharide derivative having a succinimide residue introduced in the side chain or the second liquid containing polyamine or a derivative thereof is applied to the substrate, and then the other liquid is applied or A method of gelation by dipping in a liquid to produce a gel-like reaction product is usually used, but a method of mixing and applying the above two liquids simultaneously, or a mixed application method such as a spray method is also used. obtain.
In addition, in the case of an anti-adhesion film that does not use a base material, it may be allowed to react while preventing moisture from drying after being cast in a container having a surface that is easy to peel, while mixing well with a spray, whipper, homogenizer, etc. .

The succinimide group-containing chain polysaccharide derivative is used after being dissolved in an aqueous solvent, particularly clean water such as water for injection, or a mixed solution of water and ethanol. One may be contacted with a crosslinking agent in a lyophilized solid state.
Arbitrary salts that do not adversely affect the reaction may be contained, and the pH may be adjusted with, for example, phosphate, citrate, carboxylate and the like. The preferred pH is neutral, pH 6.5 to 7.7, more preferably pH 7.3 to 7.6. The gelation reaction is basically a crosslinking reaction in which a plurality of amino groups in a polyallylamine molecule react with an end carbonyl group of a plurality of polysaccharide derivative molecules (a carbonyl group in YCO in formula (I)). (This is accompanied by elimination of N-hydroxysuccinimide).

(D) In the amidation present invention, it was found that the blood compatibility is remarkably improved by converting the -NH ₂ group remaining after gelation finally amide. Water absorption decreases as the number of carbon atoms of ethyl, propyl, butyl and alkyl groups used for acylation increases. In the case of an acylated polyallylamine, the reaction is suppressed by steric hindrance due to the introduced substituent, and therefore the acetyl group having the smallest volume is preferable. Thus, acetylation is the best way to reduce amino residues. Alternatively, the amino group of the polyamine may be acylated in advance up to 99 mol% and then used for crosslinking. In this case, the higher the acylation rate, the lower the cross-linking speed and the lower the cross-linking density, so that a fragile and quick-degrading gel is formed. Amidation can be accomplished by the action of an acylating agent.
The acylating agent is not particularly limited as long as it can convert an amino group into an amide, and examples thereof include N-acetoxysuccinimide, N-propionyloxysuccinimide, N-butyryloxysuccinimide and the like. These acylating agents are used as a solution of about 0.1 to 5% by mass in an aqueous solvent or an organic solvent, and converted to an amide by contacting with a gel or a gel-base complex described later. The conversion to amide can be confirmed by infrared rays or the like, but the reaction is usually carried out at a temperature of about 15 to 40 ° C. for 1 to 72 hours, so that the residual amino group is substantially, that is, 90% or more, preferably 95 % Or more, more preferably 98% or more. Prior to the amidation, treatments such as washing with water and dehydration may be performed.

（式中、ｐは１〜５の整数である。）で表される基と
（ｂ）：

（式中、Ｒ³は前記定義と同じ。）で表される基が挙げられるが、（ａ）の場合、多糖の水酸基−ＯＨの少なくとも一個を慣用の方法により、−Ｏ（ＣＨ₂）_pＣＯＯＨに転換するか、市販のカルボキシル化多糖を用い、これをＮ−ヒドロキシスクシンイミドと反応させる。 (E) Production method The gel composition of the present invention substantially comprises a chain polysaccharide derivative component introduced into a succinimide residue and a polyamine or a derivative thereof as described above. A chain polysaccharide derivative component in which a succinimide residue is introduced into the side chain can be produced as follows according to the preferred type of the succinimide residue.
Examples of preferred succinimide-containing R ¹ groups include
(A):

(Wherein p is an integer of 1 to 5) and (b):

(Wherein, R ³ is the same. As defined above) can be mentioned groups represented by, the method in the case, at least one conventional hydroxyl -OH polysaccharides (a), -O (CH _{2) p} Convert to COOH or use a commercially available carboxylated polysaccharide, which is reacted with N-hydroxysuccinimide.

  The esterification reaction between a carboxylated polysaccharide and N-hydroxysuccinimide can generally be performed in an aqueous solution. The amount ratio of polysaccharide to N-hydroxysuccinimide depends on the amount of carboxyl group contained in the polysaccharide, but usually 0.25 mol or more of N-hydroxysuccinimide is used with respect to the carboxyl group. The esterification reaction between the carboxylated polysaccharide and N-hydroxysuccinimide is preferably performed in the presence of a water-soluble carbodiimide. The presence of the water-soluble carbodiimide significantly accelerates the esterification reaction.

  Examples of the water-soluble carbodiimide in the present invention include 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, 1-cyclohexyl-3- (2-morpholin-4-ethyl) carbodiimide, or a hydrochloride thereof, or A sulfonate and the like can be preferably used. The reaction time of the esterification reaction is not particularly limited, but is preferably 1 minute to 48 hours. Moreover, although reaction temperature is not specifically limited, 0-40 degreeC is preferable. By reacting in the presence of 1-hydroxylbenzotriazole (HOBt) in addition to water-soluble carbodiimide, an ester compound is produced more efficiently. The amount of soluble carbodiimide and / or 1-hydroxylbenzotriazole (HOBt) added is about equimolar.

In the case of (b), it can be produced by various methods. Typically, at least one hydroxyl group -OH of the polysaccharide is first converted to -OCONHCH (R ³ ) COOR ⁵ (R ³ is as defined above). R ⁵ is a group defined in the same manner as R ³ ) to form a polysaccharide amino acid carbamate derivative, which is then deesterified and further esterified with N-hydroxysuccinimide. Is preferred.

A method for producing a polysaccharide amino acid carbamate derivative is described in Japanese Patent Application Laid-Open No. 2003-277401 by the present inventors. That is, an amino acid ester isocyanate represented by OCN-HCH (R ³ ) COOR ⁵ is reacted with a polysaccharide in the presence of lithium chloride or the like. Here, the amino acid ester isocyanate is alanine ester isocyanate and the like, and various essential amino acids and conventional derivatives thereof can be used as the amino acid portion as described above. But the manufacturing method of a polysaccharide amino acid carbamate derivative is not limited to the method of Unexamined-Japanese-Patent No. 2003-277401.

  The deesterification reaction of the polysaccharide amino acid carbamate derivative can be performed, for example, by reacting a dilute aqueous sodium hydroxide solution or dilute potassium hydroxide aqueous solution in a dilute alkaline aqueous solution. The esterification reaction between the carboxylated polysaccharide and N-hydroxysuccinimide after the deesterification reaction is the same as described above, and can generally be performed in an aqueous solution. The amount ratio of polysaccharide to N-hydroxysuccinimide depends on the amount of carboxyl group contained in the polysaccharide, but usually 0.25 mol or more of N-hydroxysuccinimide is used with respect to the carboxyl group. Moreover, an ester compound is efficiently produced | generated by making it react in presence of soluble carbodiimide and / or 1-hydroxyl benzotriazole (HOBt) similarly to the above.

The solution after completion of the reaction may be purified by removing low molecular weight substances by dialysis or ultrafiltration. The obtained aqueous solution is adjusted to a predetermined concentration or freeze-dried and redissolved at the time of use.
The concentration is appropriately adjusted according to the processing method. When the molecular weight is 125,000 to 250000, the range of 3 to 12% by mass is preferable. If it is 3% by mass or less, the viscosity and reaction rate are too low and unstable, and gelation is slow. On the other hand, when the content is 12% by mass or more, the viscosity is high and uniform processing is difficult. Most preferably, it is 5-10 mass%.
In this case, the higher the acylation rate, the lower the cross-linking speed and the lower the cross-linking density, so that a fragile and quick-degrading gel is formed. In the case of an acylated polyallylamine, the reaction is suppressed by steric hindrance due to the introduced substituent, and therefore the acetyl group having the smallest bulk is preferable.
Moreover, a gel composition-base material composite material can be manufactured by performing these reactions on a base material. For example, in the case of an artificial blood vessel, a porous cylindrical base material is immersed in the first substance, that is, an aqueous solution of a succinimidylated chain polysaccharide derivative, and a sufficient amount of voids are formed by repeated decompression and pressurization. Fulfill. Remove excess solution.

Next, for example, it is immersed in a 20% by mass aqueous solution of 20% by mole of polyallylamine and allowed to stand for 24 hours. After sufficiently washing with water to remove the unreacted crosslinking agent, the remaining amino group was acetylated by immersing in an alcohol solution of N-acetoxystacinimide for 48 hours. The acetylation conditions can be confirmed and determined by surface infrared for each individual case. Not only the surface but also the amino group inside the layer can be quantified by quantifying the terminal amino group.
Thereafter, it is thoroughly washed and dried. If necessary, a known moisturizing agent, especially glycerin having no safety problem, is added and dried to obtain the desired product.
As the substrate, a polyester resin or a fluororesin that does not biodegrade is suitable. Specific examples include polyethylene terephthalate, polybutylene terephthalate, polycyclohexane terephthalate, polyethylene 2,6-naphthalate, polybutylene 2,6-naphthalate and the like, and these are used alone or as a mixture. Among these, polyethylene terephthalate alone or a resin composition containing polyethylene terephthalate as a main component and other polyesters is preferable. As fluororesin, tetrafluoroethylene resin (PTFE) is mainly used, but as other fluororesin, tetrafluoroethylene / hexafluoroethylene copolymer (FEP), tetrafluoroethylene / perfluoroalkoxy are used. Ethylene copolymer (PFA), ethylene trifluoride chloride (PCTFE), ethylene / tetrafluoroethylene copolymer (ETFE), and the like can also be used. The shape of these base materials is not particularly limited, and is tubular, sheet-like, porous, or a combination thereof. Further, in order to increase the affinity with the hydrogel, it may be pre-processed in advance so as to reduce the contact angle with water. Examples of such processing include plasma processing.

If you want to prevent adhesions,
(a) Mix the two liquids and pour them into the mold.
(b) After one liquid is poured into the mold, the other liquid is poured.
(c) Spray the two liquids onto the mold by spray mixing spray.
The gel which does not contain a base material can be shape | molded by methods, such as.
When it is desired to accelerate decomposition (disintegration) other than in the case of an artificial blood vessel, the object can be achieved by mixing and cross-linking a succinimide derivative of carboxymethyl amylose.
The base-gel complex or gel obtained as described above reduces blood coagulation by amidating substantially all of the remaining amino groups by acylation as described above.

This property is also important in the case of preventing adhesion without using a substrate. Anti-adhesion is caused by coagulation of the bleeding blood, so when the wound surface is covered after surgery, it is shielded from contact with organs and tissues by coagulation and does not cause blood clots on the surface. It is important to do so.
Furthermore, it is required that the graft collapse after one month when the wound surface is fully healed. For this purpose, a chain polysaccharide that is easily degraded in the living body, such as amylose and dextran, may be mixed or used alone.

[Applicable to]
The gelling material for medical use of the present invention comprises (i) adhesion, blood coagulation such as artificial blood vessels, cardiovascular repair anti-adhesion membranes, patches, artificial dura mater, artificial pericardium, etc. (Iii) It is excreted so as not to cause any damage to the organ, and (vi) it has an effective property that does not cause any adverse effects on the whole body or local area in terms of biological safety. Applies to implants.

Hereinafter, the present invention will be specifically described with reference to Examples and Comparative Examples.
These do not limit the invention. In the following examples, “degree of substitution” is the degree of substitution of saccharide hydroxyl groups calculated from the results of elemental analysis.

Example 1: Synthesis of carboxymethylcellulose succinimide ester A carboxymethylcellulose succinimide ester was produced by a reaction schematically represented by the following formula (in the following formula, the acetyl group induced at the 6-position was 1/2 Although esterified, this is for convenience of explanation, and does not necessarily indicate that the reaction proceeds at this position and ratio).

Carboxymethylcellulose (25.8 g, degree of substitution 1.2, molecular weight 250,000 daltons, Aldrich) dissolved in water (400 ml of water for injection), N-hydroxysuccinimide (11.6 grams, manufactured by Junsei Chemical Co., Ltd.), 1-hydroxybenzotriazole (13.5 g, manufactured by Kanto Chemical Co., Inc.) and 1-ethyl-3- (3-dimethylaminopropyl-carbodiimide) (15.5 g, manufactured by Peptide Institute, Inc.) were added and stirred at room temperature for 48 hours. After completion of the reaction, the product was dialyzed in distilled water for 2 days with a hemodialyzer (fraction molecular weight: about 40,000, manufactured by Nikkiso Co., Ltd.). After dialysis, lyophilization gave carboxymethylcellulose succinimide ester (21.5 g).
The reaction was confirmed by IR spectrum. From the results of elemental analysis, succinimide had a degree of substitution of 0.8 per glucose unit.
In a sample bottle, the carboxymethylcellulose succinimide ester (1 g) obtained in Example 1 was dissolved in a phosphate buffer solution (20 ml, pH 7.4).
An artificial blood vessel substrate having a pleated tubular body made of polyester fiber having a length of 10 cm and an inner diameter of 10 mm was sufficiently immersed in the solution, and then the excess liquid was removed. Next, it was gently immersed in a 20% by mass polyallylamine aqueous solution and allowed to react at room temperature for 24 hours. The sufficiently washed blood vessel was dipped in an aqueous glycerin solution and dried. When the residual amino group was quantified, about 60 mol% of the polyallylamine remained without being involved in crosslinking. The artificial blood vessel was replaced with a length of 5 cm in the descending aortic region of an adult dog weighing 10 kg. Two weeks later, the blood was sacrificed and removed, and a red thrombus adhered to the entire inner surface of the artificial blood vessel.

Before the artificial blood vessel prepared in exactly the same manner was immersed in the glycerin aqueous solution, it was immersed in ethanol and replaced several times to remove water from the gel. Next, the remaining amino group was acetylated by reacting in a 1% by mass ethanol aqueous solution of N-acetoxysuccinimide for 48 hours.
After sufficiently washing with water to remove the low molecular weight compound, glycerin was used to impart moisture retention and drying.
The artificial blood vessel was sterilized with ethylene oxide gas and replaced with the descending aorta of the dog in the same manner. As a result of excision after 2 weeks, no red thrombus was adhered to the inner surface.
The artificial blood vessel was cut into 1 cm ² squares to make small pieces, which were placed subcutaneously on the rat's back and removed at 1, 2, 4, and 6 months. As a result of pathological observation, most of the gel disappeared after 6 months. Although the kidney and lung were observed, no abnormal findings were observed.

Example 2:
The carboxymethylcellulose succinimide ester prepared in Example 1 was similarly made into an aqueous solution and impregnated into a warp knitted fabric made of polyester fiber having a thickness of 0.5 ml. In order to sufficiently infiltrate the voids of the fiber, the defoaming operation was performed by repeatedly reducing pressure and pressing. Excess solution was removed. It was immersed in 75 mol% acetylated polyallylamine (molecular weight 4000) and reacted for 24 hours. The remaining amino acid was 18 mol%. After washing with water and substituting with ethanol for dehydration, it was immersed in a 1% ethanol solution of N-acetoxysuccinimide and acetylated for 48 hours. Thereafter, it was sufficiently washed with water, treated with glycerin, dried and sterilized. Some were not acetylated after cross-linking, but were moisturized with glycerin and dried.

A 1 cm ² sample of both acetylated and non-acetylated prepared in this manner was sewn as a patch to the abdominal aorta wall of the dog. As a result of excision after 2 weeks, the acetylated sample had no red thrombus adhesion and showed good performance as a cardiovascular patch. However, the blood contact surface of the patch that was not acetylated covered about 50% of the area with a red thrombus. In both cases, the gel was swollen and thickened, but the degree of swelling was lower in the gel acetylated after crosslinking. It was confirmed that the sheet containing the gel that was not acetylated had high water absorption and poor antithrombotic properties.

Example 3:
Using carboxymethylcellulose (27 g, substitution degree 1.4, molecular weight 125,000 Dalton, Nippon Paper Chemicals Co., Ltd.), the reaction was carried out in the same manner as in the previous example to obtain 22 g of carboxymethylcellulose succinimide ester. From the results of elemental analysis, succinimide was 0.65 per glucose unit.
After dialysis, the freeze-dried polymer was dissolved to make a 7% by mass aqueous solution. 10 ml of this aqueous solution was taken, and then 2 ml of 20% by mass of polyallylamine having a molecular weight of 3000 was sprayed onto a flat container with a commercially available atomized mixed spray (Meiji Machine Co., Ltd.).
When the solution of the succinimide substituted product of carboxymethylcellulose sodium salt was mixed with a small amount of methylene blue and sprayed, it was confirmed that the whole was homogeneously mixed. The reaction was performed at 40 ° C. for 24 hours.

A translucent hydrogel was obtained when peeled from the mold in water. The unreacted crosslinking agent was removed by washing with sufficient water. Half was dried and sterilized after glycerin treatment. The other half was dehydrated with alcohol and then acetylated as in the previous example. It was sterilized after washing with water, applying glycerin and drying.
The two kinds of gel sheets obtained by cutting the obtained sheet into a size of 2 × 2 cm were affixed to the spleen that had been bleeding in the abdominal cavity of the dog. Two weeks later, an autopsy revealed that the acetylated gel had almost no adhesion with the surroundings, whereas the non-acetylated sample swelled with blood infiltrating the gel and swelled firmly on the surface of the spleen. It was glued.
A 1.0 mass% physiological saline solution of carboxymethylcellulose sodium salt having a molecular weight of 500,000 daltons and 125,000 daltons was prepared and sterilized at 121 ° C. Rats were administered by intravenous injection at a rate of 50 ml / kg body weight and urine was collected for 72 hours thereafter. The results of quantitative analysis of carboxymethylcellulose in the urine by GPC are listed in the table.

これらの結果から、球状の蛋白の分画分子量では４万ダルトンの腎臓から鎖状の糖鎖は分子量が２５万ダルトン以下では吸収されずに容易に排泄されることが明らかとなった。

From these results, it was clarified that the chain sugar chain is easily excreted from the kidney of 40,000 daltons without being absorbed at a molecular weight of 250,000 daltons or less from the kidney of 40,000 daltons with the molecular weight cutoff of the globular protein.

Example 4:
Succinimide esterification was carried out in the same manner using carboxymethyl amylose sodium salt (molecular weight 500,000 Dalton, substitution degree 0.5). As a result of elemental analysis, the degree of substitution was 0.2 per glucose unit.
10 g of this was added to 100 ml of water, heated to 50 ° C. and stirred. Although it was not completely dissolved, it was poured into a flat container and lyophilized.
Next, a solution of 5,000 daltons 20% by mass of polyallylamine dissolved in alcohol was sprayed on the resulting dry sponge having a thickness of 5 mm. The reaction was allowed to proceed for 24 hours at room temperature, and then repeatedly washed with alcohol to remove unreacted crosslinking agent and reaction product.
This porous sponge was lyophilized immediately after partly immersed in water to reduce the ethanol concentration. The remainder was acetylated with the above N-acetoxysuccinimide dissolved in alcohol for 48 hours.

  Next, an artificial dura mater sheet (sold by Cosmotech Co., Ltd.) made by stretching Teflon (registered trademark) to be porous was hydrophilized by atmospheric pressure plasma treatment to increase the wettability to water. This sheet, which was impregnated with water on one side of the sheet and then adhered to the two types of sponges, was sewn on the surface of the spleen of a bleeding dog, and the appearance after 2 weeks was observed. As a result, the acetylated sheet was not fused at all, whereas in the sheet that was not subjected to the acetylation treatment after the fact, the sheet was firmly fused and could not be peeled off from the spleen surface.

As described in detail above, in the present invention, (a) a succinimide esterified polysaccharide derivative having a molecular weight of 250,000 or less,
(I) After polyamine (preferably polyallylamine) is directly or after acylating a part of amino groups, after reacting with (a) to form a gel,
(C) By artificially acylating all remaining amino groups, anti-thrombotic and biodegradable and artificial blood vessels and medical sheets that have the property of safely treating the degradation products due to excretion from the kidney I can get it.
The degree of crosslinking can be reduced by pre-acylating the polyamine to increase the degradability. As a result, it is possible to provide artificial blood vessels and other transplant materials that do not cause thrombus or fusion even by directly contacting the circulating blood or the mucous membrane in the body without using human or animal-derived raw materials.

Claims (18)

  A gel composition characterized by amidating a free amino residue that was not involved in gelation in a hydrogel obtained by reacting a chain polysaccharide derivative having a succinimide residue introduced into the side chain with polyamine or a derivative thereof .   The gel composition according to claim 1, wherein the chain polysaccharide derivative having a succinimide residue introduced in the side chain contains one succinimide per at least 20 sugar units. A chain polysaccharide derivative in which a succinimide residue is introduced into the side chain, the hydrogen atom of the hydroxyl group of the sugar is represented by the following formula (I):

The gel composition according to claim 1 or 2, wherein the gel composition is substituted with a succinimide-containing group represented by the formula (Y represents an optionally substituted alkylene or carbamoyl group). A chain polysaccharide derivative having a succinimide residue introduced in the side chain is represented by the following formula (II):

(In the formula, R ¹ is a hydrogen atom, a metal atom, an optionally substituted aliphatic hydrocarbon group or aromatic hydrocarbon group having 1 to 30 carbon atoms, or the following formula (I):

(Wherein Y represents an optionally substituted alkylene or carbamoyl group) or a succinimide-containing group represented by the following formula (III):

(In the formula, Y is as defined above, and R ² represents a hydrogen atom, a metal atom, an optionally substituted aliphatic hydrocarbon group or an aromatic hydrocarbon group having 1 to 30 carbon atoms.) X is —OR ¹ , —NHR ¹ (wherein R ¹ is the same as defined above) or a carbamoyl group, and R ^{1 in} the molecule may be the same or different. However, at least one of them is a succinimide-containing group of the above formula (I). The gel composition according to claim 3, which is a sugar chain containing a repeating unit as a repeating unit. The main structure of a chain polysaccharide derivative having a succinimide residue introduced in the side chain is represented by the following formula (IV):

The gel composition according to claim 4, wherein R ¹ and X are as defined above, and n is an integer of 3 or more. Y in the chain polysaccharide derivative is alkylene having 1 to 3 carbon atoms or the following formula (V);

(Wherein R ³ represents a hydrogen atom, or an optionally substituted aliphatic hydrocarbon group or aromatic hydrocarbon group having 1 to 30 carbon atoms). The gel composition according to any one of the above.   The gel composition according to any one of claims 1 to 6, wherein the polyamine is a polyallylamine having at least 1 mol% or more free amino groups in a side chain.   The gel composition according to any one of claims 1 to 7, wherein the polyamine has a molecular weight of 500 to 10,000.   The gel composition according to any one of claims 1 to 8, wherein the polysaccharide derivative has a molecular weight of 250,000 or less.   The gel composition-base material composite material which has the gel composition in any one of Claims 1-9 in a base material.   The gel composition-substrate composite material according to claim 10, wherein the substrate is tubular, sheet-like, porous, or a combination thereof.   The gel composition-base material composite material according to claim 10, wherein the base material is selected from a polyester resin and a fluororesin.   A transplant material that directly contacts circulating blood or a mucous membrane in the body, comprising the gel composition according to any one of claims 1 to 9 or the gel composition-substrate composite material according to any one of claims 10 to 12.   An artificial blood vessel comprising the transplant material according to claim 13.   A blood compatible organ repair patch, artificial pericardium, artificial dura mater, anti-adhesion membrane or artificial skin comprising the transplant material according to claim 13.   The patch for cardiovascular repair which is the patch for organ repair of Claim 15.   Applying one of the first liquid containing a chain polysaccharide derivative having a succinimide residue introduced into the side chain or the second liquid containing polyallylamine or a derivative thereof, and then applying or spraying or dipping the other liquid. A method for producing a gel composition-base material composite material comprising a step of reacting a residual amino group with an acylating agent after coating a material with a gel. The method for producing a gel composition-substrate composite material according to claim 17, wherein the acylating agent is an acetylating agent.