An endosomal toll-like receptor inhibitor drug with therapeutic effects in autoimmune diseases. An endosomal toll-like receptor inhibitor drug with therapeutic effects in autoimmune diseases

Autoimmune disease is a systemic inflammatory response of immune blood cells caused by self-tolerance dysfunction, due to genetic or environmental factors or a combination thereof. Studies on autoimmune disease regarding the inhibition of toll-like receptor (TLR)7/9-MyD axis or the regulation of plasmacytoid dendritic cells (pDCs) and Th17 comprising an abundance of respective receptors have shown improvements in phenotypes or molecular markers, providing clues to the practicality of the inhibitors of endosomal TLRs. This study discovered several small-molecule compounds in endosomes exerting specific inhibitory effects on SITE1 shared among TLR 3, 7, 8, and 9, named endosomal TLR inhibitors (ETIs). The drug discovery process involved identifying lead compounds optimized to the receptors using a quantitative structure-activity relationship model and subsequent modification of the compounds to improve their binding affinity to target proteins. To propose a method of verifying the drug effects against side or placebo effect arising from false positives, the drug evaluation process converged from cellular response to signaling process to protein binding. Drug values on the inhibition of endosomal TLRs in autoimmune disease were assessed based on the onset of mild-to-life-threatening autoimmune disease including psoriasis, systemic lupus erythematosus, and multiple sclerosis. Complete healing was achieved for psoriasis through direct or indirect association with TLRs. Therapeutic potential of the novel ETIs for psoriasis was verified and is considered an effective drug for other autoimmune diseases associated with the TLR7/9-MyD axis. Overall design: Gene expression profiling analysis of RNA-seq data to verify the effect of ETI41 or ETI60 treatment in TLR7 activation situation

自身免疫性疾病（Autoimmune disease）是由自身免疫耐受功能紊乱引发的免疫血细胞全身性炎症反应，发病诱因涵盖遗传因素、环境因素，或二者协同作用。针对Toll样受体（Toll-like receptor, TLR）7/9-MyD轴的抑制，或对浆细胞样树突状细胞（plasmacytoid dendritic cells, pDCs）与富含特异性受体的辅助性T细胞17（Th17）进行调控的自身免疫病相关研究，已在表型与分子标志物层面取得改善效果，为内体Toll样受体抑制剂的实用化研发提供了重要线索。本研究在内体组分中发现了数种对TLR3、7、8、9共有的SITE1位点具有特异性抑制活性的小分子化合物，将其命名为内体Toll样受体抑制剂（endosomal TLR inhibitors, ETIs）。该药物的研发流程包括：利用定量构效关系模型筛选适配靶受体的先导化合物，并通过后续结构修饰优化化合物与靶蛋白的结合亲和力。为建立可验证药物规避假阳性引发的副作用或安慰剂效应的方法，本研究的药物评价体系从细胞应答、信号转导过程延伸至蛋白质结合层面。研究基于银屑病、系统性红斑狼疮、多发性硬化等从轻症到危及生命的自身免疫病的发病情况，评估了ETIs对自身免疫病中内体Toll样受体的抑制活性。其中，银屑病可通过与TLR的直接或间接关联实现完全治愈，本研究验证了新型ETIs对银屑病的治疗潜力，且认为其可作为其他与TLR7/9-MyD轴相关的自身免疫病的有效治疗药物。整体实验设计：通过对RNA-seq数据进行基因表达谱分析，验证ETI41或ETI60在TLR7激活场景下的治疗效果。