Programmed cell death (PCD), including apoptosis, apoptotic necrosis, and pyroptosis, is involved in various organ dysfunction syndromes. Recent studies have revealed that a substrate of caspase-3, gasdermin E (GSDME), functions as an effector for pyroptosis; however, few inhibitors have been reported to prevent pyroptosis mediated by GSDME. Here, we developed a class of GSDME-derived inhibitors containing the core structure of DMPD or DMLD. Ac-DMPD-CMK and Ac-DMLD-CMK could directly bind to the catalytic domains of caspase-3 and specifically inhibit caspase-3 activity, exhibiting a lower IC₅₀ than that of Z-DEVD-FMK. Functionally, Ac-DMPD/DMLD-CMK substantially inhibited both GSDME and PARP cleavage by caspase-3, preventing apoptotic and pyroptotic events in hepatocytes and macrophages. Furthermore, in a mouse model of bile duct ligation that mimics intrahepatic cholestasis-related acute hepatic failure, Ac-DMPD/DMLD-CMK significantly alleviated liver injury. Together, this study not only identified two specific inhibitors of caspase-3 for investigating PCD but also, more importantly, shed light on novel lead compounds for treating liver failure and organ dysfunctions caused by PCD.

程序性细胞死亡 (PCD)，包括细胞凋亡、凋亡坏死和细胞焦亡，与各种器官功能障碍综合征有关。最近的研究表明，caspase-3 的底物 gasdermin E (GSDME) 是细胞焦亡的效应物。然而，据报道很少有抑制剂可以防止 GSDME 介导的细胞焦亡。在这里，我们开发了一类包含 DMPD 或 DMLD 核心结构的 GSDME 衍生抑制剂。Ac-DMPD-CMK 和 Ac-DMLD-CMK 可以直接结合 caspase-3 的催化结构域并特异性抑制 caspase-3 活性，表现出较低的 IC ₅₀与 Z-DEVD-FMK 相比。从功能上讲，Ac-DMPD/DMLD-CMK 显着抑制了 Caspase-3 对 GSDME 和 PARP 的切割，从而防止了肝细胞和巨噬细胞的凋亡和细胞焦亡事件。此外，在模拟肝内胆汁淤积相关急性肝衰竭的胆管结扎小鼠模型中，Ac-DMPD/DMLD-CMK 显着减轻了肝损伤。总之，这项研究不仅确定了两种用于研究 PCD 的 caspase-3 特异性抑制剂，而且更重要的是，揭示了治疗 PCD 引起的肝功能衰竭和器官功能障碍的新型先导化合物。