JPS6087225A - Production of fibrination promoting material - Google Patents

Abstract

PURPOSE:To obtain the titled material capable of promoting the formation of stabilized fibrins stable for a long term, by neutralizing a structure, e.g. a fiber assembly or a sponge, made from oxidized cellulose as a material, and immobilizing a specific factor and thrombin, therein. CONSTITUTION:A structure having a shape of a fiber assembly (preferably having 10-30% -CH2OH oxidized into -COOH in the cellulose for improved bioabsorbability), sponge, powder, monofilament, film or microcapsule,. A blood coagulation factor XIII (F XIII) and thrombin are bonded covalently and immobilized in the above-mentioned structure by a method of using a dehydration condensing agent, etc. to give the aimed fibrination promoting material. The resultant material is usable for treating a wound of cut or abrasion, etc., wound surface by a burn or operation wound surface or being an embolic material used for blood vessel obstruction therapy or injection material used for puncture therapy.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides an embolic material used for treatment of wounds such as cuts and abrasions, wounds caused by burns, 1 surgical side, 1 ulcers generated on the body surface, tooth extraction sockets, etc., or vascular occlusion therapy. or regarding injection materials used in puncture therapy.

More specifically, the present invention relates to a fibrin formation promoting material that can effectively promote the production of stabilized fibrin for a long period of time.

The present inventors have developed a blood coagulation box xm factor (hereinafter [FXI[I
It is abbreviated as J. ) and tronds are better materials for wound treatment because they can promote the formation of stabilized fibrin compared to materials that only have tronds immobilized, that is, materials that only promote the formation of non-stabilized fibrin. I discovered that the
5-58163) continued their research into using oxidized cellulose, which is commonly used as a hemostatic agent, as a material, and found that if oxidized cellulose was used as it was for immobilizing FXIIT and tropin, stabilized fibrin It was found that this had a negative effect on the promotion of production.

In view of the current situation, in order to solve the above-mentioned problems, the present inventors continued to study and found that, after neutralizing oxidized cellulose, FXII and thrombin were immobilized to promote the formation of stabilized fibrin. The present invention was achieved based on the discovery that the performance was well maintained.

In other words, in the present invention, after neutralizing a structure made of oxidized cellulose and having properties such as a fiber aggregate, sponge, powder, monofilament 1, film, or microcapsule, FXII and thrombin are added to the structure. This is a method for producing a fibrin formation-promoting material that can effectively promote the production of stabilized fibrin for a long period of time, which is characterized by immobilization.

Up to now, there have been studies on neutralizing oxidized cellulose and then using it as an immobilization carrier.
No. 517,772 discloses that deactivation of thrombin can be prevented by partially neutralizing oxidized cellulose before treating it with thrombin. However, there is no precedent for treating oxidized cellulose with FXIII (5), and there is also no precedent for considering the ability to promote the formation of stabilized tubulin as in the present invention.

What is the oxidized cellulose used in the present invention?2 For example, cellulose materials such as wood pulp, cotton, cotton linters, ramie, jute paper, hemp, regenerated cellulose, and rayon are mixed with ozone,
Hydrogen peroxide, periodate.

In the present invention, -CI+201 of cellulose is oxidized by a known method using nitrogen dioxide or the like.
Those in which 10 to 30% of one group is oxidized to -COO11 groups are preferably used because of their good bioabsorption, and 15 to 25
% is particularly preferably used.

In the present invention, such oxidized cellulose is used after being neutralized, and neutralization here means -COO of oxidized cellulose.
II means converting the acid group into a salt. To neutralize the oxidized cellulose, the oxidized cellulose may be treated with a hydroxide of a metal such as sodium or calcium, or with various salts such as sodium, calcium, or ammonium, such as an acetate salt or a carbonate salt. If the degree of neutralization is too high, the structure will become water-soluble and will be difficult to immobilize (on the other hand, if it is less than 3, it will be difficult to obtain good sustained release properties, and the structure will become gel-like and lose its shape). It is best to do this before you lose it.

As a guideline, 111 when neutralized oxidized cellulose is immersed in water is 3 to 7. It is preferable to carry out neutralization so that the ratio is preferably between 5 and 6.

Neutralization may be performed on the structure, or the material may be neutralized before being processed into the structure, and then the structure may be formed.

Trond used in the present invention is a protease that can convert fibrinogen to fibrin. Although thrombin is degraded from the blood of humans, cows, pigs, etc., it is preferable to use human thrombin when it is applied to the treatment of a single patient.

FXIII used in the present invention is called a fibrin stabilizing factor, and is a factor that directly acts on non-stabilized fibrin and is involved in the generation of isopeptide bonds between tubulin molecules. F
XI is isolated from blood or placenta of humans, cows, etc.2
When applied to human treatment, it is preferable to use human-derived FXnI.

FXI and thrombin can be immobilized by binding or adsorption to neutralized oxidized cellulose. To combine 1, for example 0. ZaborsK
y+″In+mobilized Enzymes″
Conventionally known covalent bonding or ionic bonding methods such as those described in C11CPress, 1973 can be employed.

To covalently bond FXIII and thrombin to neutralized oxidized cellulose, dehydration condensation of dicyclohexylcarposiimide, 1-cyclohexyl-3-(2-morpholinoethyl)-carpoimidomethotluenesulfonate, etc. Method using agent.

Oxidized cellulose is treated with polyaldehydes such as glutaraldehyde and dialdehyde starch.

A method of introducing a formyl group and then treating with FXIII and thrombin, an iminocarbonate group, a bromoacetyl group using bromic cyan instead of polyaldehyde, an acid anhydride group using polymaleic anhydride such as a maleic anhydride copolymer, and toluene diisocyanate. Isocyanate groups by polyisocyanates such as xamethylene diisocyanate.

Ethylene glycol, tetramethyl glycol.

Since an epoxy group can be introduced into a structure using a polyglycidyl derivative such as glycerin, and a chlortriazinyl group can be introduced into a structure using cyanuric chloride, this is a method for binding FXI and thrombin using functional groups that form these covalent bonds. etc. can be adopted.

6- In order to ionically bond FXIU and thrombin to neutralized oxidized cellulose, the carboxyl group of oxidized cellulose may be used, or as mentioned above, formyl group, iminocarbonate group, bromoacetyl group, acid anhydride group, isocyanate group.

Oxidized cellulose into which epoxy groups and chlortriazinyl groups have been introduced can be aminated by treatment with polyamines such as polyethyleneimine, and carboxylated by treatment with aminocarboxylic acids such as 2-glycine. FXl and thrombin can be ionically bonded to a structure into which amino groups and carboxyl groups, which are ion exchange groups, have been introduced.

When FXI and thrombin are adsorbed onto neutralized oxidized cellulose by a physical adsorption method, an entrapment method, etc., it can be carried out as follows. That is, by dissolving FXTI and thrombin in a solvent that can wet neutralized oxidized cellulose, and treating the structure with this solution, FX
IIr and thrombin can be physically adsorbed. The entrapment method is a method in which FX■ and thrombin are wrapped in a fine lattice of gel so that they cannot be released.

When immobilizing FXIII and thrombin by any of the above methods, FXI and thrombin may be immobilized at the same time, or FXI may be immobilized first and then thrombin may be immobilized, The opposite may be true. Alternatively, FXIII-immobilized products and thrombin-immobilized products may be manufactured separately, and these may be uniformly mixed or layered.

9 In addition, when manufacturing the material of the present invention, FXI
Calcium ions involved in the activation of [[ can be immobilized. Furthermore, when producing the material of the present invention, pharmaceuticals such as antiplasmin agents, bactericides, antibiotics, hormones, etc. can be immobilized on the structure together with FXI and thrombin, if necessary. antiplasmin.

It is an inhibitor of plasmin, a fibrin-dissolving enzyme5
Therefore, inhibiting plasmin suppresses fibrinolytic activity, ie, fibrinolytic activity. Therefore, a material in which antiplasmin is immobilized along with FXI and thrombin can promote fibrin production by suppressing fibrinolytic activity. Examples of antiplasmin include aprotinin extracted from bovine lungs, natural substances such as bebustin isolated from microbial culture fluid, leupepsin, antipain, and chymostatin, ε-aminocaproic acid, tranexamic acid,
Examples include gabexate mesylate, and particularly preferred are ε-aminocaproic acid and tranexamic acid.

The material of the present invention can be used to protect wounds such as cuts and abrasions, ulcers generated on the surface of wounds caused by burns, tooth extraction sockets, anastomoses, etc., or as an embolization material used for vascular occlusion therapy, and for puncture therapy. It is used in the injection material used and can effectively promote the production of stabilized fibrin for a long period of time. Therefore 1
When the material of the present invention is used as a wound protection agent,
It has a hardening effect that significantly accelerates wound healing and at the same time relieves local pain, and when used as an embolic agent, it quickly and firmly forms a thrombus, preventing blood vessel recanalization and preventing injection materials from forming. When used as a drug, it enables rapid and reliable hemostasis.

Hereinafter, the present invention will be explained in more detail by showing two examples. As thrombin, human plasma thrombin from Midori Juji Co., Ltd. was used, and as FXI, FXI concentrated dry preparation was used. FXII [Preparation contains 4 fresh human plasma per bottle]
FXI has an activity corresponding to 60 times the amount present in ml.

In addition, a test for the ability to form stabilized fibrin was conducted as follows. That is, Ca
After FXIII was activated by treatment with an aqueous solution containing th, the structure was placed in a 0.5% fibrinogen aqueous solution and left at 37° C. for 10 minutes. Thereafter, it was confirmed whether the fibrin generated on the surface of the structure was stabilized fibrin. Since unstabilized fibrin dissolves in 1% monochloroacetic acid, but stabilized fibrin does not dissolve in 1% monochloroacetic acid, the fibrin formed as described above

Claims (1)

[Claims] (Fiber aggregates, sponges, powders, monofilaments, films made of 11-oxide cellulose. After neutralizing a structure having a shape such as a microcapsule, a blood coagulation box xm factor is added to the structure. and a method for producing a fibrin formation promoting material capable of effectively promoting the production of stabilized fibrin for a long period of time, which is characterized by immobilizing thrombin.