Site-Specific PEGylation of HR2 Peptides: Effects of PEG Conjugation Position and Chain Length on HIV-1 Membrane Fusion Inhibition and Proteolytic Degradation
Site-Specific PEGylation of HR2 Peptides: Effects of PEG Conjugation Position and Chain Length on HIV-1 Membrane Fusion Inhibition and Proteolytic Degradation
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DOI：
10.1021/bc3002248
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发表时间：
2012-08-01
影响因子：
4.7
通讯作者：
Klok, Harm-Anton
中科院分区：
文献类型：
--
作者：
Danial, Maarten;van Dulmen, Tim H. H.;Klok, Harm-Anton
来自HIV-1包膜糖蛋白gp 41的HR 1或HR 2区的肽已被证明是防止病毒-宿主细胞膜融合的有效抑制剂。然而，这些肽类药物的血浆半衰期相对较短，并且易受酶降解的影响。修饰的肽/蛋白质与聚（乙二醇）（PEG）是一个行之有效的策略，以克服这些限制：这份手稿提出了一个系统的研究结果的影响的网站HR 2衍生肽的PEG化，以及PEG分子量对这些共轭物的生物活性和蛋白水解稳定性。在基于模型细胞-细胞的测定中对缀合物的融合抑制功效的研究揭示了与野生型HR 2衍生肽相比，PEG化肽的活性损失。然而，通过控制PEG化位点，更具体地说，通过在肽的非相互作用α-螺旋表面的沿着更中心的位置之一引入PEG链，可以使活性损失最小化。在基于胰蛋白酶的模型测定中评估PEG-肽缀合物的蛋白水解稳定性，其揭示降解半衰期增加高达3.4倍，这可能有助于补偿PEG-肽缀合物与野生型肽相比的较低抑制功效。这项研究的结果强调了位点特异性聚乙二醇化的力量，以提高肽/蛋白质药物的稳定性，同时最大限度地减少对生物活性的不利影响。
Peptides derived from the HR1 or HR2 regions of the HIV-1 envelope glycoprotein gp41 have been shown to be effective inhibitors to prevent virus-host cell membrane fusion. These peptide drugs, however, suffer from relatively short plasma half-lives and are susceptible to enzymatic degradation. Modification of peptides/proteins with poly (ethylene glycol) (PEG) is a well established strategy to overcome these limitations: This manuscript presents the results of a systematic study on the influence of the site of PEGylation of HR2-derived peptides, as well as of PEG molecular weight on the biological activity and proteolytic stability of these conjugates. Investigation of the fusion inhibitory efficacy of the conjugates in a model cell-cell based assay revealed a loss in activity for the PEGylated peptides as compared to the wild-type HR2-derived peptide. The loss of activity, however, can be minimized by controlling the site of PEGylation, more specifically, by introducing the PEG chain at one of the more central positions along the non interacting alpha-helical surface of the peptides. The proteolytic stability of the PEG-peptide conjugates was assessed in a trypsin-based model assay, which revealed an up to 3.4-fold increase in degradation half-life that may help to compensate for the lower inhibitory efficacy of the PEG-peptide conjugates as compared to the wild type peptide. The results of this study emphasize the power of site specific PEGylation to improve the stability of peptide/protein drugs while,minimizing adverse effects on biological activity.