Zhang Min, Dou Huaizhi, Yin Lichen, Zhang Yu, Zhu Siyu, Yin Chunhua
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, PR China.
Pharmazie. 2009 Mar;64(3):190-6.
Insulin was modified with monomethoxypoly(ethylene glycol) 5000 (MPEG5000) by reaction of insulin amino groups with activated MPEG5000, and mono-, di- and tri-PEGylated insulin were obtained. Circular dichroism demonstrated that the attachment of MPEG5000 to insulin did not alter the tertiary structure of insulin. PEGylation of insulin could inhibit the self-association tendency of insulin, which decreased with the increase of substitution degree. The physical stability and proteolytic stability of insulin increased after MPEG conjugation, and the stabilities of the conjugates depended on the PEGylation degree. In vivo biological activity of mono-PEGylated insulin following parenteral administration both in mice and rats was preserved while di- and tri-PEGylated insulin showed a great loss in biological activity. In addition, mono-PEGylated insulin administered subcutaneously in mice as well as intravenously in rats displayed extended action profiles. These results indicated that mono-substituted MPEG5000-insulin conjugate was a good candidate for insulin replacement therapy.
通过胰岛素氨基与活化的甲氧基聚乙二醇5000（MPEG5000）反应对胰岛素进行修饰，得到了单聚乙二醇化、双聚乙二醇化和三聚乙二醇化的胰岛素。圆二色性表明MPEG5000与胰岛素的连接未改变胰岛素的三级结构。胰岛素的聚乙二醇化可抑制胰岛素的自缔合倾向，且该倾向随取代度的增加而降低。MPEG偶联后胰岛素的物理稳定性和蛋白水解稳定性增加，且缀合物的稳定性取决于聚乙二醇化程度。单聚乙二醇化胰岛素经皮下注射给小鼠以及经静脉注射给大鼠后，其体内生物活性得以保留，而双聚乙二醇化和三聚乙二醇化胰岛素的生物活性则大幅丧失。此外，单聚乙二醇化胰岛素经皮下注射给小鼠以及经静脉注射给大鼠后显示出延长的作用曲线。这些结果表明单取代的MPEG5000 - 胰岛素缀合物是胰岛素替代治疗的良好候选物。
