State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, China.
Department of Pharmacy, The Second Affiliated Hospital of Jiaxing University, China.
J Cell Mol Med. 2021 Nov;25(22):10534-10542. doi: 10.1111/jcmm.16984. Epub 2021 Oct 10.
The main mechanism of pyroptosis is Caspase-1-mediated GSDMD cleavage, and GSDMD is also the executive protein of pyroptosis. Our previous study has shown that mafenide can inhibit pyroptosis by inhibiting the GSDMD-Asp275 site to suppress cleavage. In this study, sulfonamide was used as the parent nucleus structure to synthesize sulfa-4 and sulfa-20. Screening of drug activity in the pyroptosis model of BV2 and iBMDM cell lines revealed the efficacy of five compounds were superior to mafenide, which exerted a better inhibitory effect on the occurrence of pyroptosis. For in vivo assay, Sulfa-4 and Sulfa-22 were intervened in the neuroinflammation APP/PS1 mice. As a result, the administration of Sulfa-4 and Sulfa-22 could significantly inhibit the activation of microglia, decrease the expression of inflammatory factors in the central nervous system and simultaneously suppress the production of p30-GSDMD as well as the expression of upstream NLRP3 inflammasome and Caspase-1 protein. Immunoprecipitation and Biotin-labelled assay confirmed the targeted binding relationship of Sulfa-4 and Sulfa-22 with GSDMD protein in the iBMDM model in vitro. In this study, we investigated a new type inhibitor of GSDMD cleavage, which exerted a good inhibitory effect on pyroptosis and provided new references for the development of inflammatory drugs in the future.
细胞焦亡的主要机制是 Caspase-1 介导的 GSDMD 切割，GSDMD 也是细胞焦亡的执行蛋白。我们之前的研究表明，苯扎氯铵通过抑制 GSDMD-Asp275 位点抑制切割来抑制细胞焦亡。在这项研究中，磺胺作为母核结构合成了磺胺-4 和磺胺-20。在 BV2 和 iBMDM 细胞系的细胞焦亡模型中筛选药物活性，发现五种化合物的疗效优于苯扎氯铵，对细胞焦亡的发生有更好的抑制作用。在体内实验中，磺胺-4 和磺胺-22 干预 APP/PS1 神经炎症小鼠。结果表明，磺胺-4 和磺胺-22 的给药可显著抑制小胶质细胞的激活，降低中枢神经系统中炎症因子的表达，同时抑制 p30-GSDMD 的产生以及上游 NLRP3 炎性体和 Caspase-1 蛋白的表达。免疫沉淀和生物素标记实验证实了磺胺-4 和磺胺-22 在体外 iBMDM 模型中与 GSDMD 蛋白的靶向结合关系。本研究探索了一种新型 GSDMD 切割抑制剂，对细胞焦亡有较好的抑制作用，为未来炎症药物的开发提供了新的参考。
