Lee D J, Cameron A J, Wright T H, Harris P W R, Brimble M A
School of Chemical Sciences , The University of Auckland , 23 Symonds St , Auckland 1142 , New Zealand . Email:
School of Biological Sciences , The University of Auckland , 3 Symonds St , Auckland 1142 , New Zealand.
Chem Sci. 2018 Oct 29;10(3):815-828. doi: 10.1039/c8sc03409e. eCollection 2019 Jan 21.
The clinical significance of batch-wise variability on the pharmacokinetics and potency of commercial erythropoietin (EPO), prepared recombinantly as a heterogeneous mixture of glycoforms, necessitates the development of synthetic strategies to afford homogenous EPO formulations. Herein we present a previously unexplored and divergent route towards 'click' neoglycoprotein analogues of EPO, employing one-pot native chemical ligation (NCL) of alkynylated peptides and copper-catalysed azide-alkyne cycloaddition (CuAAC) with azido monosaccharides. By design, our synthetic platform permits glycosylation at virtually any stage, providing flexibility for the synthesis of various glycoforms for biological analysis. Insights obtained from attempted folding of our 'click' neoglycoprotein EPO analogue, bearing four different neutral sugar moieties, highlight the important role played by the charged oligosaccharides present in native EPO glycoproteins.
作为糖型异质混合物重组制备的商业促红细胞生成素（EPO），其批次间变异性对药代动力学和效力的临床意义，使得开发合成策略以提供均质EPO制剂成为必要。在此，我们展示了一条前所未有的、不同的合成路线，用于制备EPO的“点击”新糖蛋白类似物，该路线采用炔基化肽的一锅法天然化学连接（NCL）以及与叠氮单糖的铜催化叠氮-炔环加成反应（CuAAC）。通过设计，我们的合成平台允许在几乎任何阶段进行糖基化，为合成用于生物学分析的各种糖型提供了灵活性。从我们带有四种不同中性糖部分的“点击”新糖蛋白EPO类似物的折叠尝试中获得的见解，突出了天然EPO糖蛋白中存在的带电荷寡糖所起的重要作用。
