小分子AI44靶蛋白过氧化物还原酶1的发现及其抑制NLRP3炎症小体活化的分子机制

研究表明NLRP3炎症小体介导固有免疫反应对炎症及相关疾病的发生至关重要。前期从系列化合物中发现AI44可高效抑制NLRP3炎症小体活化，对小鼠败血症有较好改善效果，初步证实其靶蛋白为过氧化物还原酶1（PRDX1）。本项目拟从分子、细胞及动物水平研究AI44通过作用于PRDX1抑制NLRP3活化、改善炎症的效果及机制：用免疫共沉淀、免疫荧光、SPR、ITC等多种手段确证AI44与PRDX1结合及其构效关系；用计算机模拟、肽段截短和点突变等确定AI44与PRDX1的结合位点；揭示AI44对PRDX1氧化还原酶活、寡聚化、磷酸化、泛素化及胞内定位的影响并考察AI44通过PRDX1抑制NLRP3炎症小体的分子机制；在败血症和结肠炎模型中探讨AI44的治疗效果。本项目的实施预期为基于NLRP3炎症小体的疾病干预提供药物靶标，为炎症及相关疾病的治疗提供新的思路。

Studies have proved that innate immunity and inflammation-reaction mediated by NLRP3 inflammasome play essential roles during the development of inflammation and related disease. Previously we have screened AI44 as an effective candidate for NLRP3 inflammasome inhibition and found that it was benefit for improvement of sepsis in mice. Several evidence suggested its target was Peroxiredoxin1. The project here aims to clarify the character of the AI44-PRDX1 interaction and detailed mechanism for AI44’s NLRP3 inhibition effect from the molecular, cellular and animal levels, demonstrating the role of this mechanism involved in AI44’s anti-inflammation activity：IP, IF, SPR, ITC etal confirm the interaction, chemical structure and pharmacologic activity relationship between AI44 and PRDX1; computer docking, peptide truncation, point mutation for searching the binding site between AI44 and PRDX1; uncovering the effect of AI44 on enzyme activity, oligomerization, phosphorylation, ubiquitin, sub-cellular localization of PRDX1 and the detail mechanism for AI44-driven PRDX1 mediated NLRP3 inhibition; Employ the mice sepsis, colitis disease model for testing the therapeutic effect of AI44. Results of this study will be helpful to provide candidates for the therapy of inflammation and related disease by targeting PRDX1-NLRP3 inflammasome and, more importantly, may help to identify some critical proteins for regulation of NLRP3 inflammasome.