PEGylated glucagon-like peptide-1 displays preserved effects on insulin release in isolated pancreatic islets and improved biological activity in db/db mice
PEGylated glucagon-like peptide-1 displays preserved effects on insulin release in isolated pancreatic islets and improved biological activity in db/db mice
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DOI：
10.1007/s00125-006-0234-3
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发表时间：
2006-07-01
期刊：
影响因子：
8.2
通讯作者：
Lee, KC
中科院分区：
文献类型：
--
作者：
Lee, S;Youn, YS;Lee, KC
目的/假设：胰高血糖素样肽-1（GLP-1）通过二肽基肽酶-IV和中性内肽酶24.11的快速降解和清除是GLP-1作为潜在降糖药开发的主要障碍。在这项研究中，设计了新的酶抗性聚乙二醇（PEG）结合GLP-1类似物，并检查了代谢稳定性和生物学效价。材料和方法：制备两种单聚乙二醇化GLP-1类似物，N-末端修饰的N-PEG/GLP-1和Lys-修饰的Lys-PEG/GLP-1。在血浆和组织浸提液中检测稳定性。使用离体大鼠胰岛进行体外胰岛素释放研究，而在db/db小鼠中测量体内血药反应。结果：Lys-PEG/GLP-1在血浆、肝和肾匀浆中的半衰期分别是GLP-1的40、10和28倍。Lys-PEG/GLP-1以剂量和葡萄糖依赖性方式刺激胰岛中的胰岛素分泌，并且与GLP-1一样有效。相比之下，N-PEG/GLP-1显示出延长的代谢稳定性，但具有显著较低的生物活性。Lys-PEG/GLP-1（9 nmol/kg i.p.）对于未禁食的db/db小鼠，GLP-1稳定了血糖（p < 0.001），而GLP-1（9 nmol/kg）仅引起葡萄糖水平的微小变化。在禁食db/db小鼠的OGTT期间，以1、3和9 nmol/kg（i. p.）与安慰剂相比，GLP-1（9 nmol/kg）分别使葡萄糖AUC（0- 3 h）降低48.7 +/- 9.4、55.0 +/- 2.9和63.4 +/-2.5%（p < 0.01），而GLP-1（9 nmol/kg）使葡萄糖水平降低39.5 +/- 12.9%（p < 0.01）。结论/解读：本研究证明，位点特异性聚乙二醇化GLP-1类似物对降解具有抗性。这些类似物的增强的生物效力突出了它们作为新的GLP-1样降糖药的潜力。
Aims/hypothesis: The rapid degradation and clearance of glucagon-like peptide-1 (GLP-1) by the enzymes dipeptidyl peptidase-IV and neutral endopeptidase 24.11 are the main impediments to the development of GLP-1 as a potential glucose-lowering agent. In this study, new enzyme-resistant polyethylene glycol (PEG)-conjugated GLP-1 analogues were designed and examined for metabolic stability and biological potency. Materials and methods: Two mono-PEGylated GLP-1 analogues, N-terminally modified N-PEG/GLP-1 and Lys-modified Lys-PEG/GLP-1, were prepared. Stability was tested in plasma and tissue extracts. In vitro insulin release studies were performed using isolated rat pancreatic islets, while in vivo glycaemic responses were measured in db/db mice. Results: The half-life of Lys-PEG/GLP-1 was 40-, 10- and 28-fold longer than that of GLP-1 in plasma, liver and kidney homogenates, respectively. Lys-PEG/GLP-1 stimulated insulin secretion in the islets in a dose- and glucose-dependent manner, and was as potent as GLP-1. In contrast, N-PEG/GLP-1 showed extended metabolic stability but had significantly lower biological activity. The administration of Lys-PEG/GLP-1 (9 nmol/kg i.p.) to non-fasted db/db mice stabilised glycaemia (p < 0.001), whereas GLP-1 (9 nmol/kg) only caused small changes in glucose level. During OGTT in fasted db/db mice, Lys-PEG/GLP-1 administered at 1, 3 and 9 nmol/kg (i.p.) reduced the glucose AUC(0-3h) by 48.7 +/- 9.4, 55.0 +/- 2.9 and 63.4 +/- 2.5%, respectively, compared with placebo (p < 0.01), whereas GLP-1 (9 nmol/kg) lowered the glucose level by 39.5 +/- 12.9% (p < 0.01). Conclusions/interpretation: This study demonstrates that site-specific PEGylated GLP-1 analogues are resistant to degradation. The enhanced biological potencies of these analogues highlight their potential as new, GLP-1-like glucose-lowering agents.