Identification of a Small-Molecule Entry Inhibitor for Filoviruses
Identification of a Small-Molecule Entry Inhibitor for Filoviruses
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DOI：
10.1128/jvi.01456-10
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发表时间：
2011-04-01
影响因子：
5.4
通讯作者：
Bowlin, Terry L.
中科院分区：
文献类型：
--
作者：
Basu, Arnab;Li, Bing;Bowlin, Terry L.
埃博拉病毒（EBOV）引起严重的出血热，对此没有治疗选择。防止EBOV进入宿主细胞是一种有吸引力的抗病毒策略，FDA批准的抗HIV药物恩夫韦肽已经验证了这一点。为了鉴定EBOV进入的抑制剂，使用EBOV包膜糖蛋白（EBOV-GP）基因来产生假型病毒以用于筛选化学文库。从利用假型病毒的小分子化合物文库的高通量筛选（HTS）中鉴定苯并二氮杂卓衍生物（化合物7）。化合物7在基于细胞的测定中被验证为感染性EBOV和马尔堡病毒（MARV）的抑制剂，50%抑制浓度（IC（50））分别为10 μ M和12 μ M。添加时间和结合研究表明，化合物7在EBOV感染期间的早期阶段与EBOV-GP结合。初步的Schrodinger SiteMap计算，使用已发表的EBOV-GP晶体结构在其融合前构象，建议在GP 1和GP 2界面处或附近的疏水口袋作为化合物7结合的合适位点。该预测得到突变分析的支持，表明残基Asn 69、Leu 70、Leu 184、Ile 185、Leu 186、Lys 190和Lys 191对于化合物7及其类似物与EBOV-GP的结合是关键的。我们假设化合物7与该疏水口袋结合，并因此抑制EBOV感染细胞，但机制的细节仍有待确定。总之，我们已经确定了一系列新型苯二氮卓类化合物，它们适合优化作为丝状病毒感染的潜在抑制剂。
Ebola virus (EBOV) causes severe hemorrhagic fever, for which therapeutic options are not available. Preventing the entry of EBOV into host cells is an attractive antiviral strategy, which has been validated for HIV by the FDA approval of the anti-HIV drug enfuvirtide. To identify inhibitors of EBOV entry, the EBOV envelope glycoprotein (EBOV-GP) gene was used to generate pseudotype viruses for screening of chemical libraries. A benzodiazepine derivative (compound 7) was identified from a high-throughput screen (HTS) of small-molecule compound libraries utilizing the pseudotype virus. Compound 7 was validated as an inhibitor of infectious EBOV and Marburg virus (MARV) in cell-based assays, with 50% inhibitory concentrations (IC(50)s) of 10 mu M and 12 mu M, respectively. Time-of-addition and binding studies suggested that compound 7 binds to EBOV-GP at an early stage during EBOV infection. Preliminary Schrodinger SiteMap calculations, using a published EBOV-GP crystal structure in its prefusion conformation, suggested a hydrophobic pocket at or near the GP1 and GP2 interface as a suitable site for compound 7 binding. This prediction was supported by mutational analysis implying that residues Asn69, Leu70, Leu184, Ile185, Leu186, Lys190, and Lys191 are critical for the binding of compound 7 and its analogs with EBOV-GP. We hypothesize that compound 7 binds to this hydrophobic pocket and as a consequence inhibits EBOV infection of cells, but the details of the mechanism remain to be determined. In summary, we have identified a novel series of benzodiazepine compounds that are suitable for optimization as potential inhibitors of filoviral infection.