蚕蛹壳聚糖甘氨酸衍生物的制备及其抗凝血作用研究

Preparation of Chitosan Glycine Derivative from Silkworm Pupae and its Anti-coagulant Activity

壳聚糖是生产抗凝血物质的理想原料。采用化学方法对蚕蛹壳聚糖进行分子修饰,即在蚕蛹壳聚糖分子上引入甘氨酸和环氧氯丙烷反应的中间体,制备壳聚糖甘氨酸衍生物。通过红外光谱及核磁共振分析其分子结构,结果显示甘氨酸已接枝到蚕蛹壳聚糖分子上,为在C6-OH发生醚化反应的壳聚糖甘氨酸衍生物。抗凝血试验结果表明,蚕蛹壳聚糖甘氨酸衍生物具有抗凝血活性,当其质量浓度达到2mg/mL时抗凝血效果最佳,可使血液完全不凝固。从凝血时间、凝血酶原时间、活化部分凝血活酶时间、凝血酶时间的检测结果可知,蚕蛹壳聚糖甘氨酸衍生物影响到外源性、内源性凝血系统和凝血活性酶的形成。依据抗凝血试验结果分析认为,蚕蛹壳聚糖甘氨酸衍生物中C6引入的羧基与壳聚糖C2上的氨基共同作用,产生了抗凝血效果。研究结果提示:蚕蛹壳聚糖甘氨酸衍生物可进一步开发为医用抗凝血材料。

Chitosan is an ideal material for preparing anti-coagulant substances.In present study,chemical methods were employed to conduct molecular modifications towards chitosan of silkworm pupae.Glycine and epichlorohydrin reaction intermediates were introduced onto the chitosan molecules to prepare the glycine derivative of chitosan.Infrared spectrometry and nuclear magnetic resonance analyses revealed that glycines had been grafted onto chitosan molecules of silkworm pupae.The resultant glycine derivative of chitosan was from the etherification reaction on C6-OH.Anti-co-agulation test indicated that chitosan glycine derivative of silkworm pupae had anti-coagulant activity.When its mass concentration reached 2 mg/mL,the blood could be maintained non-coagulant.Determination on coagulation time,throm-binogen time,partial activation time of thromboplastin,and thrombin time revealed that chitosan glycine derivative of silkworm pupae could exert influences on the formation of exogenous and endogenous coagulation systems and expression of enzymes with coagulation activity.The anti-coagulant test results suggest that the C6 carboxyl groups introduced onto chitosan glycine derivative of silkworm pupae and the chitosan＇s C2 amino groups had worked together to produce the anti-coagulant effect.The obtained results show that the chitosan glycine derivative of silkworm pupae could be further developed into anti-coagulant materials for medical use.