Monoconjugation of Human Amylin with Methylpolyethyleneglycol

Amylin is a pancreatic hormone cosecreted with insulin that exerts unique roles in metabolism and glucose homeostasis. The therapeutic restoration of postprandial and basal amylin levels is highly desirable in diabetes mellitus. Protein conjugation with the biocompatible polymer polyethylene glycol (PEG) has been shown to extend the biological effects of biopharmaceuticals. We have designed a PEGylated human amylin by using the aminoreactive compound methoxylpolyethylene glycol succinimidyl carbonate (mPEGsc). The synthesis in organic solvent resulted in high yields of monoPEGylated human amylin, which showed large stability against aggregation, an 8 times increase in half-life in vivo compared to the non-conjugated amylin, and pharmacological activity as shown by modulation of cAMP production in MCF–7 cell line, decrease in glucagon and modulation of glycemia following subcutaneous administration in mice. Altogether these data reveal the potential use of PEGylated human amylin for the restoration of fasting amylin levels.

胰淀素（Amylin）是一类与胰岛素共同分泌的胰腺激素，在代谢调控与血糖稳态维持中发挥独特生理功能。糖尿病患者亟需实现餐后与基础胰淀素水平的治疗性恢复。将蛋白质与生物相容性聚合物聚乙二醇（polyethylene glycol, PEG）偶联，已被证实可有效延长生物制药制剂的生物学效应时长。本研究通过氨基反应性化合物甲氧基聚乙二醇琥珀酰亚胺碳酸酯（methoxylpolyethylene glycol succinimidyl carbonate, mPEGsc），设计制备了聚乙二醇化人胰淀素。在有机溶剂中进行合成可获得高收率的单聚乙二醇化人胰淀素，该产物具备优异的抗聚集稳定性，其体内半衰期较非偶联胰淀素提升8倍；同时通过MCF-7细胞系中环磷酸腺苷（cAMP）生成的调控实验、小鼠皮下给药后胰高血糖素水平降低及血糖稳态调控实验，证实了其药理活性。综上，上述研究数据揭示了聚乙二醇化人胰淀素在恢复空腹胰淀素水平方面的应用潜力。