combinatorial computational approaches to identify tetracycline derivatives as flavivirus inhibitors组合计算方法来确定四环素衍生物为黄病毒抑制剂.pdfVIP

Combinatorial Computational Approaches to Identify Tetracycline Derivatives as Flavivirus Inhibitors 1,2 1 1 1 1 Jinn-Moon Yang , Yan-Fu Chen , Yu-Yin Tu , Kuei-Rong Yen , Yun-Liang Yang * 1 Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, 2 Institute of Bioinformatics, National Chiao Tung University, Hsinchu, Taiwan Limited structural information of drug targets, cellular toxicity possessed by lead compounds, and large amounts of potential leads are the major issues facing the design-oriented approach of discovering new leads. In an attempt to tackle these issues, we have developed a process of virtual screening based on the observation that conformational rearrangements of the dengue virus envelope protein are essential for the mediation of viral entry into host cells via membrane fusion. Screening was based solely on the structural information of the Dengue virus envelope protein and was focused on a target site that is presumably important for the conformational rearrangements necessary for viral entry. To circumvent the issue of lead compound toxicity, we performed screening based on molecular docking using structural databases of medical compounds. To enhance the identification of hits, we further categorized and selected candidates according to their novel structural characteristics. Finally, the selected candidates were subjected to a biological validation assay to assess inhibition of Dengue virus propagation in mammalian host cells using a plaque formation assay. Among the 10 compounds examined, rolitetracycline and doxycycline significantly inhibited plaque formation, demonstrating their inhibitory effect on dengue virus propagation. Both compounds were tetracycline derivatives with IC50s estimated to be 67.1 mM and 55.6 mM, respectively. Their docked conformations disp