Fibrinogen-like globe domain of human Tenascin-C (hFBG-C); A Target Enabling Package

Chronic activation of the innate immune system by the damage-associated molecular pattern FBG-C (C-terminal fibrinogen-like globe domain of Tenascin-C) contributes to a variety of inflammatory diseases including arthritis, systemic sclerosis, and cancer. This TEP summarizes the first reported efforts to develop small-molecule FBG-C binders, with the aim to disrupt FBG-C-mediated pro-inflammatory protein-protein interactions (PPIs). We present the soluble expression of disulphide-containing human FBG-C (hFBG-C) in <em>E. coli</em>, the novel structure of hFBG-C, and preliminary chemical matter against hFBG-C derived from a crystallographic fragment screen. Finally, we introduce two robustly validated cellular assays, in either immortalized monocytes or primary human macrophages, which provide a route to development of small molecules which inhibit hFBG-C-activated inflammation.损伤相关分子模式（damage-associated molecular pattern）FBG-C（腱生蛋白-C（Tenascin-C）的C端纤维蛋白原样球状结构域）介导的先天免疫系统慢性激活，可引发关节炎、系统性硬化症及癌症等多种炎症性疾病。本技术实验方案（TEP）总结了首例报道的开发小分子FBG-C结合剂的相关工作，旨在阻断FBG-C介导的促炎蛋白-蛋白相互作用（protein-protein interactions, PPIs）。我们实现了含二硫键的人源FBG-C（hFBG-C）在大肠杆菌（E. coli）中的可溶性表达，解析了hFBG-C的全新晶体结构，并获得了通过晶体学片段筛选得到的靶向hFBG-C的初步化学实体。最后，我们介绍了两种分别基于永生化单核细胞与原代人巨噬细胞的经过严格验证的细胞检测体系，可为开发抑制hFBG-C激活的炎症反应的小分子化合物提供可行途径。