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.
NU6300是一种通过共价结合Gasdermin D (GSDMD)蛋白C191位点来特异性抑制细胞焦亡的小分子化合物[1]。NU6300能够与细胞焦亡的关键执行蛋白GSDMD的第191位半胱氨酸（C191）发生共价反应，阻断GSDMD的切割和棕榈酰化，从而阻止其N端片段在细胞膜上形成孔隙，抑制细胞焦亡的发生[2]。
在体外，NU6300(0.5–10μM)预处理人单核细胞THP-1、小鼠骨髓来源巨噬细胞及HEK-293T细胞40分钟至16小时，随后通过尼日利亚霉素（10μM; 1h）、poly(dA:dT)（1μg/mL; 6h）或鞭毛蛋白（1μg/mL; 6h）等激活AIM2、NLRC4或NLRP3炎症小体，显著抑制GSDMD的切割和棕榈酰化，阻断其膜定位及寡聚化，从而减少细胞焦亡和促炎因子IL-1β的释放[3]。
在体内，NU6300(5mg/kg)腹腔注射，用于处理DSS诱导的结肠炎小鼠模型（连续五天）或LPS诱导的脓毒症小鼠模型（单次注射）。NU6300显著减轻DSS诱导的结肠炎小鼠的体重下降、疾病活动指数升高、结肠缩短和组织病理损伤，改善上皮和黏膜损伤、隐窝扩张和杯状细胞耗竭。NU6300显著提高脓毒症小鼠存活率，降低脾脏指数及促炎细胞因子水平[3]。