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Preparation method and application of oxidized low-molecular-weight heparin-antithrombin compounds

A low-molecular-weight heparin and antithrombin technology, applied in the preparation of oxidized low-molecular-weight heparin-antithrombin complex-coated extracorporeal circulation pipelines, in the field of blood and biocompatible coatings, which can solve problems that cannot be completely prevented. Blood activation, plasma protein adsorption is not selective, etc., to achieve the effect of increasing affinity, reducing adsorption, and reducing consumption

Inactive Publication Date: 2010-11-10
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0006] Aiming at the technical defects that the heparin-coated extracorporeal circulation pipeline still has no selectivity for plasma protein adsorption and cannot completely prevent blood activation mediated by surgical wounds, the present invention prepares an oxidized low molecular weight heparin-antithrombin complex, And use polyethyleneimine-glutaraldehyde combination technology to immobilize it on the surface of the extracorporeal circulation pipeline to further improve the biocompatibility of the extracorporeal circulation pipeline

Method used

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  • Preparation method and application of oxidized low-molecular-weight heparin-antithrombin compounds
  • Preparation method and application of oxidized low-molecular-weight heparin-antithrombin compounds
  • Preparation method and application of oxidized low-molecular-weight heparin-antithrombin compounds

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preparation example Construction

[0017] According to the above technical principles, the present invention provides a method for preparing an oxidized low molecular weight heparin-antithrombin complex, the method comprising the following steps:

[0018] (1) React excess sodium periodate and low-molecular-weight heparin sodium injection for 30 minutes at room temperature in the dark, and centrifuge three times with an AmiconUltra-4 centrifugal ultrafiltration device with a molecular weight cut-off of 3K to obtain low-molecular-weight heparin injection. Molecular weight heparin sodium solution, wherein the average molecular weight of the low molecular weight heparin sodium is 5000.

[0019] (2) Mix the oxidized low molecular weight heparin sodium solution obtained in step (1) with antithrombin in 0.02mol / L pH7.3 phosphate buffered saline for 40 ° C water bath reaction for 14 days, promote the non-enzymatic glycosylation reaction between the terminal aldose residue of oxidized low molecular weight heparin and th...

Embodiment

[0024] Example: Preparation of polyvinyl chloride pipe coated with oxidized low molecular weight heparin-antithrombin complex and its physical and chemical properties detection

[0025] Add 2g of sodium periodate and 8ml of low molecular weight heparin sodium injection into a 25ml volumetric flask, then add deionized water to 25ml, and react in the dark for 30 minutes at room temperature; Centrifuge at 6500 rpm for 20 min at room temperature, discard the ultrafiltrate, add deionized water to the retentate, and repeat the centrifugation 3 times to obtain about 2 ml of oxidized low molecular weight heparin sodium solution.

[0026] 2mg antithrombin, 0.18gNaCl, 0.44ml0.2mol / LNaH 2 PO 4 Solution, 1.56ml0.2mol / LNa 2 HPO 4 solution, oxidized low molecular weight heparin sodium solution and 16ml deionized water were fully mixed to obtain a mixed reaction solution of about 20ml, which was put into 40 ° Reaction in C water bath for 14d; Add 2ml0.5mol / L sodium cyanoborohydride sol...

Embodiment 2

[0037] Example 2: Biocompatibility Evaluation of Polyvinyl Chloride Tubing Coated with Oxidized Low Molecular Weight Heparin-Antithrombin Complex

[0038] Biocompatibility evaluation experiments were carried out with the heparin-coated, low-molecular-weight heparin-coated and oxidized low-molecular-weight heparin-antithrombin complex-coated polyvinyl chloride pipes prepared in Example 1. The experiment was divided into five groups: control group, no coating group, heparin coating group, low molecular weight heparin coating group and oxidized low molecular weight heparin-antithrombin complex coating group. After human anticoagulated whole blood was simulated in vitro with Chandlar-Loop model for 4 hours, platelet count, fibrinogen content, plasma interleukin-8 (IL-8) and monocyte chemoattractant protein- 1 (MCP-1), and observe the inner surface of each group of pipelines with a scanning electron microscope. The results showed that the platelet counts in each group (×10 9 / L...

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Abstract

The invention discloses a method for preparing oxidized low-molecular-weight heparin-antithrombin compounds and applying the oxidized low-molecular-weight heparin-antithrombin compounds to coating layer used for improving extracorporeal circulation pipeline blood and biocompatibility. The method comprises the following steps: firstly, oxidizing low-molecular-weight heparin through sodium periodate; then, carrying out nonenzymatic glycosylation reaction with antithrombin; obtaining the purified oxidized low-molecular-weight heparin-antithrombin compounds through centrifugal ultrafiltration and diethylaminoethyl-agarose high-flowing-speed anion exchange chromatography; and then, using the polymine-glutaric dialdehyde combining technology for immobilizing the low-molecular-weight heparin-antithrombin compounds on the surface of the extracorporeal circulation pipeline. The extracorporeal circulation pipeline of the oxidized low-molecular-weight heparin-antithrombin compound coating layer prepared by the invention has better coating layer stability, anticoagulation performance and biocompatibility than the extracorporeal circulation pipeline of the heparin coating layer and low-molecular-weight heparin coating layer, the inflammatory mediator generation and release and the blood coagulation system activation caused by the contact of blood with the surface of the extracorporeal circulation pipeline can be lightened.

Description

technical field [0001] The invention belongs to the field of medical technology, and relates to the preparation of an oxidized low molecular weight heparin-antithrombin complex and a coating method for improving the blood and biocompatibility of medical equipment in contact with blood, in particular to a A method for preparing an extracorporeal circulation tube coated with oxidized low molecular weight heparin-antithrombin complex. Background technique [0002] Extracorporeal circulation is a routine technique in cardiac surgery at present. Extracorporeal circulation is mainly composed of connecting pipes, oxygenator, blood reservoir, arterial microembolus filter, etc., and its constituent materials are mostly chemical substances such as polyvinyl chloride, polytetrafluoroethylene, polyester, polyethylene, silicone rubber, and stainless steel. . Due to the poor biocompatibility of these artificial synthetic materials, contact with blood can cause activation of the body's h...

Claims

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Application Information

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IPC IPC(8): A61L31/10A61L31/08A61L33/16A61L33/10
Inventor 杨剑罗鹏易定华杨丽芳曹瑞军刘维永杨春周华
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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