Mathiesen Isabel R, Calder Ewen D D, Kunzelmann Simone, Walport Louise J
Protein-Protein Interaction Laboratory, The Francis Crick Institute, London, UK.
Department of Chemistry, Molecular Sciences Research Hub, Imperial College London, London, UK.
Commun Chem. 2024 Dec 19;7(1):304. doi: 10.1038/s42004-024-01388-9.
Covalent drugs can achieve high potency with long dosing intervals. However, concerns remain about side-effects associated with off-target reactivity. Combining macrocyclic peptides with covalent warheads provides a solution to minimise off-target reactivity: the peptide enables highly specific target binding, positioning a weakly reactive warhead proximal to a suitable residue in the target. Here we demonstrate the direct discovery of covalent cyclic peptides using encoded libraries containing a weakly electrophilic cysteine-reactive fluoroamidine warhead. We combine direct incorporation of the warhead into peptide libraries using the flexible in vitro translation system with a peptide selection approach that identifies only covalent target binders. Using this approach, we identify potent and selective covalent inhibitors of the peptidyl arginine deiminase, PADI4 or PAD4, that react exclusively at the active site cysteine. We envisage this approach will enable covalent peptide inhibitor discovery for a range of related enzymes and expansion to alternative warheads in the future.
共价药物可以通过延长给药间隔时间来实现高效能。然而，与脱靶反应相关的副作用仍然令人担忧。将大环肽与共价弹头相结合提供了一种将脱靶反应降至最低的解决方案：肽能够实现高度特异性的靶点结合，将弱反应性弹头定位在靶点中合适残基的附近。在这里，我们展示了使用含有弱亲电半胱氨酸反应性氟脒弹头的编码文库直接发现共价环肽。我们将弹头通过灵活的体外翻译系统直接掺入肽库，并结合一种仅识别共价靶点结合物的肽筛选方法。使用这种方法，我们鉴定出了肽基精氨酸脱亚氨酶PADI4的强效和选择性共价抑制剂，该抑制剂仅在活性位点半胱氨酸处发生反应。我们设想这种方法将能够发现一系列相关酶的共价肽抑制剂，并在未来扩展到其他弹头。
