Non-covalent mTOR inhibition suppresses NLRP3-GSDMD inflammasome-mediated pyroptosis.

Pyroptosis is an inflammatory form of programmed cell death primarily driven by activation of the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, which triggers caspase-1-dependent cleavage of gasdermin D (GSDMD) and subsequent release of pro-inflammatory cytokines. The mechanistic target of rapamycin (mTOR) signaling pathway has emerged as a critical regulator of pyroptosis, yet its precise role in inflammasome activation remains to be incompletely understood. Here, we report that compound CD25, a novel non-covalent small-molecule inhibitor, suppresses NLRP3 inflammasome-mediated pyroptosis by targeting both mTORC1 and mTORC2 pathways. Furthermore, CD25 preserved mitochondrial integrity, thereby inhibiting apoptosis-associated speck-like protein containing a CARD (ASC) oligomerization, and blocking GSDMD cleavage and membrane pore formation in macrophages. In murine models of lipopolysaccharide-induced sepsis and acetaminophen-induced acute liver injury, administration of CD25 markedly attenuated immune-mediated inflammatory injury and ameliorated tissue damage. Collectively, these findings highlight mTOR signaling as a promising therapeutic target for modulating NLRP3-dependent pyroptosis in immune-related inflammatory disorders.