NU6300 is a small molecule compound that specifically inhibits pyroptosis by covalently binding to the C191 site of the Gasdermin D (GSDMD) protein[1]. NU6300 can covalently react with the cysteine 191 (C191) of GSDMD, a key executioner protein in pyroptosis, thereby blocking the cleavage and palmitoylation of GSDMD. This prevents the N-terminal fragment of GSDMD from forming pores in the cell membrane, ultimately inhibiting the occurrence of pyroptosis[2].
In vitro, pretreatment of human monocytic THP-1 cells, mouse bone marrow-derived macrophages, and HEK-293T cells with NU6300 (0.5–10μM) for 40 minutes to 16 hours, followed by activation of AIM2, NLRC4, or NLRP3 inflammasomes via nigericin (10μM; 1h), poly(dA:dT) (1μg/mL; 6h), or flagellin (1μg/mL; 6h), significantly inhibited the cleavage and palmitoylation of GSDMD. This blocked its membrane localization and oligomerization, thereby reducing pyroptosis and the release of the proinflammatory cytokine IL-1β[3].
In vivo, NU6300 (5mg/kg) was administered via intraperitoneal injection in DSS-induced colitis mouse models (daily for five consecutive days) or LPS-induced sepsis mouse models (single injection). NU6300 significantly alleviated body weight loss, disease activity index elevation, colon shortening, and histopathological damage in DSS-induced colitis mice, while also improving epithelial and mucosal damage, crypt dilation, and goblet cell depletion. Additionally, NU6300 significantly increased the survival rate of septic mice, reduced the spleen index, and lowered the levels of proinflammatory cytokines[3].
References:
[1] Shua Y, Wang Q, Wang Y, et al. Progress in small-molecule inhibitors of gasdermin D. Eur J Med Chem. 2025 Dec 15;300:118171.
[2] Anscombe E, Meschini E, Mora-Vidal R, et al. Identification and Characterization of an Irreversible Inhibitor of CDK2. Chem Biol. 2015 Sep 17;22(9):1159-64.
[3] Jiang X, Zhang X, Cai X, et al. NU6300 covalently reacts with cysteine-191 of gasdermin D to block its cleavage and palmitoylation. Sci Adv. 2024 Feb 9;10(6):eadi9284.