PDF(Pubmed)
Abstract：
Since NEK7 is critical for NLRP3 inflammasome activation, NEK7 inhibitors could be employed as therapeutic agents against gout, a representative disease caused by NLRP3 inflammasome.
We designed NEK7 inhibitors based on biochemical kinome profiling of 2,7-substituted thieno[3,2-d]pyrimidine derivatives (SLC3031~3035 and SLC3037). Inflammasome activation was assessed by ELISA of IL-1b and immunoblotting of IL-1b maturation after treatment of bone marrow-derived macrophages with LPS+monosodium urate (MSU). NLPR3 binding to NEK7 and oligomerization were examined using immunoprecipitation and Blue Native gel electrophoresis, respectively. In vivo effect was investigated by studying gross and histopathological changes of food pad tissue of MSU-injected mice, together with assays of maturation of IL-1b and ASC speck in the tissue.
SLC3037 inhibited inflammasome by MSU and other inflammasome activators through blockade of NLRP3 binding to NEK7 or oligomerization, and subsequent ASC oligomerization/phosphorylation. SLC3037 significantly reduced foot pad thickness and inflammation by MSU, which was superior to the effects of colchicine. SLC3037 significantly reduced content or maturation of IL-1b and ASC speck in the food pad. The number and height of intestinal villi were decreased by colchicine but not by SLC3037.
SLC3037, a NLRP3 inhibitor blocking NEK7 binding to NLRP3, could be a novel agent against diseases associated with NLRP3 inflammasome activation such as gout, cardiovascular diseases, metabolic syndrome or neurodegenerative diseases.
We designed NEK7 inhibitors based on biochemical kinome profiling of 2,7-substituted thieno[3,2-d]pyrimidine derivatives (SLC3031~3035 and SLC3037). Inflammasome activation was assessed by ELISA of IL-1b and immunoblotting of IL-1b maturation after treatment of bone marrow-derived macrophages with LPS+monosodium urate (MSU). NLPR3 binding to NEK7 and oligomerization were examined using immunoprecipitation and Blue Native gel electrophoresis, respectively. In vivo effect was investigated by studying gross and histopathological changes of food pad tissue of MSU-injected mice, together with assays of maturation of IL-1b and ASC speck in the tissue.
SLC3037 inhibited inflammasome by MSU and other inflammasome activators through blockade of NLRP3 binding to NEK7 or oligomerization, and subsequent ASC oligomerization/phosphorylation. SLC3037 significantly reduced foot pad thickness and inflammation by MSU, which was superior to the effects of colchicine. SLC3037 significantly reduced content or maturation of IL-1b and ASC speck in the food pad. The number and height of intestinal villi were decreased by colchicine but not by SLC3037.
SLC3037, a NLRP3 inhibitor blocking NEK7 binding to NLRP3, could be a novel agent against diseases associated with NLRP3 inflammasome activation such as gout, cardiovascular diseases, metabolic syndrome or neurodegenerative diseases.
摘要：
■由于NEK7对NLRP3炎性体激活至关重要，NEK7抑制剂可用作痛风的治疗剂，由NLRP3炎性体引起的代表性疾病。
■我们基于2,7-取代的噻吩并[3,2-d]嘧啶衍生物（SLC3031〜3035和SLC3037）的生化动力学谱分析设计了NEK7抑制剂。在用LPS+尿酸单钠(MSU)处理骨髓来源的巨噬细胞后,通过IL-1b的ELISA和IL-1b成熟的免疫印迹评估炎性体活化。使用免疫沉淀和蓝色天然凝胶电泳检查NLPR3与NEK7的结合和寡聚化，分别。通过研究MSU注射小鼠的食物垫组织的总体和组织病理学变化来研究体内作用，连同组织中IL-1b和ASC斑点的成熟测定。
■SLC3037通过阻断NLRP3与NEK7的结合或寡聚化而抑制MSU和其他炎症小体激活剂的炎症小体，和随后的ASC寡聚化/磷酸化。SLC3037显著降低了MSU的脚垫厚度和炎症，其效果优于秋水仙碱。SLC3037显着降低了食物垫中IL-1b和ASC斑点的含量或成熟。秋水仙素降低了肠绒毛的数量和高度，但SLC3037没有降低。
■SLC3037是一种阻断NEK7与NLRP3结合的NLRP3抑制剂，可能是一种新的药物，可以对抗与NLRP3炎性体激活相关的疾病，如痛风，心血管疾病,代谢综合征或神经退行性疾病。
■我们基于2,7-取代的噻吩并[3,2-d]嘧啶衍生物（SLC3031〜3035和SLC3037）的生化动力学谱分析设计了NEK7抑制剂。在用LPS+尿酸单钠(MSU)处理骨髓来源的巨噬细胞后,通过IL-1b的ELISA和IL-1b成熟的免疫印迹评估炎性体活化。使用免疫沉淀和蓝色天然凝胶电泳检查NLPR3与NEK7的结合和寡聚化，分别。通过研究MSU注射小鼠的食物垫组织的总体和组织病理学变化来研究体内作用，连同组织中IL-1b和ASC斑点的成熟测定。
■SLC3037通过阻断NLRP3与NEK7的结合或寡聚化而抑制MSU和其他炎症小体激活剂的炎症小体，和随后的ASC寡聚化/磷酸化。SLC3037显著降低了MSU的脚垫厚度和炎症，其效果优于秋水仙碱。SLC3037显着降低了食物垫中IL-1b和ASC斑点的含量或成熟。秋水仙素降低了肠绒毛的数量和高度，但SLC3037没有降低。
■SLC3037是一种阻断NEK7与NLRP3结合的NLRP3抑制剂，可能是一种新的药物，可以对抗与NLRP3炎性体激活相关的疾病，如痛风，心血管疾病,代谢综合征或神经退行性疾病。
