Mudhasani Rajini, Kota Krishna P, Retterer Cary, Tran Julie P, Tritsch Sarah R, Zamani Rouzbeh, Whitehouse Chris A, Bavari Sina
Therapeutic Discovery Center, Molecular and Translational Sciences Division, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD, USA.
Therapeutic Discovery Center, Molecular and Translational Sciences Division, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD, USA
J Biomol Screen. 2015 Jan;20(1):141-52. doi: 10.1177/1087057114556253. Epub 2014 Oct 23.
High-content image-based screening was developed as an approach to test a small-molecule library of compounds targeting signal transduction pathways for antiviral activity against multiple highly pathogenic RNA viruses. Of the 2843 compounds screened, 120 compounds exhibited ≥60% antiviral activity. Four compounds (E225-0969, E528-0039, G118-0778, and G544-0735), which were most active against Rift Valley fever virus (RVFV) and showed broad-spectrum antiviral activity, were selected for further evaluation for their concentration-response profile and cytotoxicity. These compounds did not show any visible cytotoxicity at the highest concentration of compound tested (200 µM). All four of these compounds were more active than ribavirin against several viruses. One compound, E225-0969, had the lowest effective concentration (EC50 = 1.9-8.92 µM) for all the viruses tested. This compound was 13- and 43-fold more inhibitory against RVFV and Chikungunya virus (CHIKV), respectively, than ribavirin. The highest selectivity index (>106.2) was for E225-0969 against CHIKV. Time-of-addition assays suggested that all four lead compounds targeted early steps in the viral life cycle (entry and/or replication) but not virus egress. Overall, this work demonstrates that high-content image analysis can be used to screen chemical libraries for new antivirals against highly pathogenic viruses.
基于高内涵图像的筛选技术被开发出来，用于测试一个针对信号转导途径的小分子化合物文库，以检测其对多种高致病性RNA病毒的抗病毒活性。在筛选的2843种化合物中，有120种化合物表现出≥60%的抗病毒活性。选择了四种对裂谷热病毒（RVFV）活性最高且具有广谱抗病毒活性的化合物（E225-0969、E528-0039、G118-0778和G544-0735），进一步评估它们的浓度-反应曲线和细胞毒性。在测试的最高化合物浓度（200µM）下，这些化合物未显示出任何明显的细胞毒性。这四种化合物对几种病毒的活性均高于利巴韦林。其中一种化合物E225-0969对所有测试病毒的有效浓度最低（EC50 = 1.9-8.92µM）。该化合物对RVFV和基孔肯雅病毒（CHIKV）的抑制作用分别比利巴韦林高13倍和43倍。E225-0969对CHIKV的选择性指数最高（>106.2）。添加时间实验表明，所有四种先导化合物均靶向病毒生命周期的早期步骤（进入和/或复制），而非病毒释放。总体而言，这项工作表明高内涵图像分析可用于筛选化学文库，以寻找针对高致病性病毒的新型抗病毒药物。
