摘要
目的:通过优化聚乙二醇(PEG)对水蛭素的化学修饰反应条件,得到具有生物学活性、半衰期延长的修饰产物。方法:通过改变化学反应中聚乙二醇与水蛭素的比例获得不同修饰度的产物;采用SDS-PAGE分析产物的修饰度;TH发色底物法分析聚乙二醇修饰前、后水蛭素的活性;用凝胶过滤层析对水蛭素的PEG修饰产物进行分离纯化;采用部分活化凝血酶时间(APTT)鉴定PEG修饰水蛭素在动物体内延长半衰期的效果。结果:化学反应中PEG与水蛭素摩尔比超过3∶1时,水蛭素被修饰后抗凝活性明显下降。凝胶过滤层析方法分离得到不同修饰程度的水蛭素。活性分析表明,连接1个和2个聚乙二醇分子的水蛭素保持了原有活性,并可不同程度地延长体内半衰期;连接3个以上PEG的水蛭素活性明显降低。结论:通过控制PEG对水蛭素的修饰反应条件,得到了具有生物学活性、体内半衰期延长的修饰产物。
Objective ： To obtain the optimal reaction condition for pegylation of hirudin in order prolong half-life of hirudin in vivo by pegylation. Methods： The pegylated products were isolated by gel filtration chromatography. Chromogenic substrate TH was used to analyze the special activity of hirudin with different pegylation degree. Activated partial thrombin time （APTT） of rabbit serum was analyzed to identify the half-life of hirudin in vivo. Results： The activity of the pegylated mixture decreased dramatically when the ratio of PEG vs hirudin exceeded 3： 1. The activities of pegylated hirudins linked with 1 or 2 PEG were well retained but that with 3 PEG obviously attenuated. The half-lives of hirudins with different pegylated degree were prolonged to different extents. Conclusion： Under the optimization of modified conditions, two pegylated hirudins with prolonged half life were obtained without activity loss.
出处
《军事医学科学院院刊》
CSCD
北大核心
2005年第4期333-336,共4页
Bulletin of the Academy of Military Medical Sciences
基金
国家"973"计划基金资助项目(2002CB713804)
关键词
水蛭素
聚乙二醇
化学修饰
半衰期
hirudin
polyethylene glycols
chemical modification
half-life
