Rhomboid-family intramembrane proteases regulate important biological processes and have been associated with malaria, cancer, and Parkinson's disease. However, due to the lack of potent, selective, and pharmacologically compliant inhibitors, the wide therapeutic potential of rhomboids is currently untapped. Here, we bridge this gap by discovering that peptidyl α-ketoamides substituted at the ketoamide nitrogen by hydrophobic groups are potent rhomboid inhibitors active in the nanomolar range, surpassing the currently used rhomboid inhibitors by up to three orders of magnitude. Such peptidyl ketoamides show selectivity for rhomboids, leaving most human serine hydrolases unaffected. Crystal structures show that these compounds bind the active site of rhomboid covalently and in a substrate-like manner, and kinetic analysis reveals their reversible, slow-binding, non-competitive mechanism. Since ketoamides are clinically used pharmacophores, our findings uncover a straightforward modular way for the design of specific inhibitors of rhomboid proteases, which can be widely applicable in cell biology and drug discovery.

中文翻译：

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设计有效、选择性和药理学相容的菱形蛋白酶抑制剂的通用和模块化策略

菱形家族膜内蛋白酶调节重要的生物学过程，并与疟疾、癌症和帕金森病有关。然而，由于缺乏有效的、选择性的和药理学兼容的抑制剂，菱形肌的广泛治疗潜力目前尚未开发。在这里，我们通过发现在酮酰胺氮处被疏水基团取代的肽基 α-酮酰胺是在纳摩尔范围内有效的菱形抑制剂，比目前使用的菱形抑制剂高出三个数量级，从而弥补了这一差距。这种肽基酮酰胺对菱形体表现出选择性，使大多数人丝氨酸水解酶不受影响。晶体结构表明，这些化合物以类似底物的方式共价结合菱形的活性位点，动力学分析揭示了它们的可逆、缓慢结合、非竞争机制。由于酮酰胺是临床上使用的药效团，我们的研究结果揭示了一种设计菱形蛋白酶特异性抑制剂的简单模块化方法，可广泛应用于细胞生物学和药物发现。