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目的我们在实验中偶然发现了一种能抑制RORγt活性的新型化合物,拟通过鉴定该化合物对Th17体外分化及其细胞因子分泌的影响对其进行生物学评价,以确定该化合物是否可作为一种新型的自身免疫病候选药物。方法构建能在Jurkat细胞中稳定表达Gal4-RORγt的荧光素酶报告系统,通过该体系发现了一种对RORγt有显著抑制活性的新型化合物,并在小鼠Th17细胞分化实验中对该化合物的作用效果进行体外验证。在此基础上,对该化合物进行生物功能分析,包括EC50、CC50的测定以及T细胞特异性分析。结果通过体外验证实验,我们确定该化合物(编号为compound 2)能显著的抑制Th17分化及其细胞因子IL-17A、IL-17F的表达和分泌。在后续的生物学评价中,我们还发现该化合物对RORγt有较高的抑制效率,较低的细胞毒性和较强的T细胞特异性。结论本研究中得到的化合物compound 2可作为一种潜在的新型前导化合物应用于治疗自身免疫性疾病和一些炎症性疾病。
Purpose We have in the experiment accidentally discovered a novel compound that inhibits RORγt activity and is intended to evaluate its biological effects by identifying the effect of this compound on Th17 in vitro differentiation and its cytokine secretion to determine whether this compound can be used as a New autoimmune disease drug candidates. METHODS: A luciferase reporter system capable of stably expressing Gal4-RORγt in Jurkat cells was constructed. A novel compound with significant inhibitory activity against RORγt was found and its effect on the differentiation of mouse Th17 cells Effect of in vitro validation. On this basis, the compounds were analyzed for biological function, including the determination of EC50 and CC50, as well as T cell specificity analysis. Results Through in vitro validation experiments, we identified that this compound (compound 2) significantly inhibited Th17 differentiation and the expression and secretion of its cytokines IL-17A and IL-17F. In the subsequent biological evaluation, we also found that the compound has higher inhibitory efficiency on RORγt, lower cytotoxicity and stronger T cell specificity. Conclusion The compound compound 2 obtained in this study can be used as a potential new lead compound in the treatment of autoimmune diseases and some inflammatory diseases.