An Ultrapotent and Selective Cyclic Peptide Inhibitor of Human β-Factor XIIa in a Cyclotide Scaffold
An Ultrapotent and Selective Cyclic Peptide Inhibitor of Human β-Factor XIIa in a Cyclotide Scaffold
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DOI：
10.1021/jacs.1c07574
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发表时间：
2021-11-01
影响因子：
15
通讯作者：
Suga, Hiroaki
中科院分区：
文献类型：
--
作者：
Liu, Wenyu;de Veer, Simon J.;Suga, Hiroaki
环肽是植物源肽，具有由其首尾相连的环状骨架和胱氨酸结核心形成的复杂结构。这些结构特征支持环肽的天然生物活性，以及它们作为药物先导物的有益特性，包括高蛋白水解稳定性和细胞渗透性。然而，它们固有的结构复杂性对环肽工程提出了挑战，特别是对于访问具有足够化学多样性的文库以设计有效和选择性的环肽变体。在这里，我们报告了一种使用mRNA展示的策略，使我们能够从包含基于木麻黄胰蛋白酶抑制剂-II（MCoTI-II）的环肽支架的超过10（12）个成员的文库中选择有效的基于环肽的FXIIa抑制剂。最有效和选择性的抑制剂cMCoFx 1对FXIIa具有pM抑制常数，相对于相关丝氨酸蛋白酶具有大于三个数量级的选择性，实现了对内源性凝血途径的特异性抑制。cMCoFx 1和FXIIa的共晶结构揭示了在接触界面的几个位置上的相互作用，这些位置传递了高亲和力结合，突出表明这种环肽是用于治疗开发的有吸引力的胱氨酸结支架。
Cyclotides are plant-derived peptides with complex structures shaped by their head-to-tail cyclic backbone and cystine knot core. These structural features underpin the native bioactivities of cyclotides, as well as their beneficial properties as pharmaceutical leads, including high proteolytic stability and cell permeability. However, their inherent structural complexity presents a challenge for cyclotide engineering, particularly for accessing libraries of sufficient chemical diversity to design potent and selective cyclotide variants. Here, we report a strategy using mRNA display enabling us to select potent cyclotide-based FXIIa inhibitors from a library comprising more than 10(12) members based on the cyclotide scaffold of Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II). The most potent and selective inhibitor, cMCoFx1, has a pM inhibitory constant toward FXIIa with greater than three orders of magnitude selectivity over related serine proteases, realizing specific inhibition of the intrinsic coagulation pathway. The cocrystal structure of cMCoFx1 and FXIIa revealed interactions at several positions across the contact interface that conveyed high affinity binding, highlighting that such cyclotides are attractive cystine knot scaffolds for therapeutic development.