Rhomboid proteases form one of the most widespread intramembrane protease families. They have been implicated in variety of human diseases. The currently reported rhomboid inhibitors display some selectivity, but their construction involves multistep synthesis protocols. Here, we report benzoxazin-4-ones as novel inhibitors of rhomboid proteases with a covalent, but slow reversible inhibition mechanism. Benzoxazin-4-ones can be synthesized from anthranilic acid derivatives in a one-step synthesis, making them easily accessible. We demonstrate that an alkoxy substituent at the 2-position is crucial for potency and results in low micromolar inhibitors of rhomboid proteases. Hence, we expect that these compounds will allow rapid synthesis and optimization of inhibitors of rhomboids from different organisms.

菱形蛋白酶形成最广泛的膜内蛋白酶家族之一。它们与多种人类疾病有关。当前报道的菱形抑制剂表现出一定的选择性，但是它们的构建涉及多步合成方案。在这里，我们报告苯并恶嗪-4-酮作为具有共价但缓慢的可逆抑制机制的菱形蛋白酶的新型抑制剂。苯并恶嗪-4-酮可从邻氨基苯甲酸衍生物一步合成，使其易于合成。我们证明，在2位上的烷氧基取代基对于效力至关重要，并导致菱形蛋白酶的低微摩尔抑制剂。因此，我们预期这些化合物将允许快速合成和优化来自不同生物体的菱形抑制剂。