Yusuf Hamied Department of Chemistry, University of Cambridge, Cambridge, UK.
Biologics Engineering, Oncology R&D, AstraZeneca, The Discovery Centre; Cambridge Biomedical Campus, Cambridge, UK.
Nat Commun. 2024 Aug 24;15(1):7308. doi: 10.1038/s41467-024-51610-4.
Cyclization provides a general strategy for improving the proteolytic stability, cell membrane permeability and target binding affinity of peptides. Insertion of a stable, non-reducible linker into a disulphide bond is a commonly used approach for cyclizing phage-displayed peptides. However, among the vast collection of cysteine reactive linkers available, few provide the selectivity required to target specific cysteine residues within the peptide in the phage display system, whilst sparing those on the phage capsid. Here, we report the development of a cyclopropenone-based proximity-driven chemical linker that can efficiently cyclize synthetic peptides and peptides fused to a phage-coat protein, and cyclize phage-displayed peptides in a site-specific manner, with no disruption to phage infectivity. Our cyclization strategy enables the construction of stable, highly diverse phage display libraries. These libraries can be used for the selection of high-affinity cyclic peptide binders, as exemplified through model selections on streptavidin and the therapeutic target αvβ3.
环化提供了一种提高肽的蛋白水解稳定性、细胞膜通透性和靶结合亲和力的通用策略。在二硫键中插入稳定的、不可还原的连接子是一种常用的环化噬菌体展示肽的方法。然而，在大量可用的半胱氨酸反应性连接子中，很少有连接子能够在噬菌体展示系统中针对肽中的特定半胱氨酸残基提供所需的选择性，同时避免噬菌体衣壳上的半胱氨酸。在这里，我们报告了一种基于环丙烯酮的近邻驱动化学连接子的开发，该连接子可以有效地环化合成肽和融合到噬菌体衣壳蛋白上的肽，并以特异性方式环化噬菌体展示的肽，而不影响噬菌体的感染力。我们的环化策略能够构建稳定的、高度多样化的噬菌体展示文库。这些文库可用于筛选高亲和力的环状肽结合物，如在链霉亲和素和治疗靶点αvβ3上的模型选择中所证明的那样。
