Abstract:
NLRP3 is an intracellular sensor protein that detects a broad range of danger signals and environmental insults. Its activation results in a protective pro-inflammatory response designed to impair pathogens and repair tissue damage via the formation of the NLRP3 inflammasome. Assembly of the NLRP3 inflammasome leads to caspase 1-dependent secretory release of the pro-inflammatory cytokines IL-1β and IL-18 as well as to gasdermin d-mediated pyroptotic cell death. Herein, we describe the discovery of a novel indazole series of high affinity, reversible inhibitors of NLRP3 activation through screening of DNA-encoded libraries and the potent lead compound 3 (BAL-0028, IC50 = 25 nM) that was identified directly from the screen. SPR studies showed that compound 3 binds tightly (KD range 104-123 nM) to the NACHT domain of NLRP3. A CADD analysis of the interaction of compound 3 with the NLRP3 NACHT domain proposes a binding site that is distinct from those of ADP and MCC950 and includes specific site interactions. We anticipate that compound 3 (BAL-0028) and other members of this novel indazole class of neutral inhibitors will demonstrate significantly different physical, biochemical, and biological properties compared to NLRP3 inhibitors previously identified.
摘要:
NLRP3是一种细胞内传感蛋白,可检测各种危险信号和环境危害。其活化导致保护性促炎反应,其设计为通过NLRP3炎性体的形成损害病原体和修复组织损伤。NLRP3炎性体的组装导致促炎细胞因子IL-1β和IL-18的胱天蛋白酶1依赖性分泌释放,并导致gasdermind介导的细胞凋亡。在这里,我们描述了一种高亲和力的新型吲唑系列的发现,通过筛选DNA编码的文库和直接从筛选鉴定的有效先导化合物3(BAL-0028,IC50=25nM),NLRP3激活的可逆抑制剂。SPR研究显示化合物3与NLRP3的NACHT结构域紧密结合(KD范围104-123nM)。化合物3与NLRP3NACHT结构域的相互作用的CADD分析提出了与ADP和MCC950的结合位点不同的结合位点,并且包括特异性位点相互作用。我们预计,化合物3(BAL-0028)和其他成员的这种新型的吲哚类中性抑制剂将表现出显著不同的物理,生物化学,和与先前鉴定的NLRP3抑制剂相比的生物学特性。