Novel Small Molecule Entry Inhibitors of Ebola Virus
Novel Small Molecule Entry Inhibitors of Ebola Virus
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DOI：
10.1093/infdis/jiv223
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发表时间：
2015-10-01
影响因子：
6.4
通讯作者：
Bowlin, Terry L.
中科院分区：
文献类型：
--
作者：
Basu, Arnab;Mills, Debra M.;Bowlin, Terry L.
背景目前的埃博拉病毒（EBOV）爆发凸显了缺乏可用的抗病毒药物或疫苗来治疗感染患者和阻止EBOV传播的问题。EBOV糖蛋白（GP）在病毒感染的早期阶段起着关键作用，包括受体结合和膜融合，使其成为开发抗EBOV药物的潜在靶点。我们报道了2种新的靶向病毒进入的EBOV抑制剂的鉴定。为了鉴定EBOV进入的小分子抑制剂，我们使用表达EBOV-GP的基于人类免疫缺陷病毒的假型病毒进行了基于细胞的高通量筛选。鉴定了两种化合物，并使用免疫荧光法、AlphaLISA法和组织蛋白酶B抑制酶测定法进行了作用机制研究。我们报告了2个新的进入抑制剂的鉴定。这些抑制剂（1）抑制EBOV感染（50%抑制浓度，约0.28和约10 μ mol/L）在进入的后期阶段，（2）诱导Niemann-Pick C表型，和（3）抑制GP-Niemann-Pick C1（NPC 1）蛋白相互作用。我们已经鉴定了2种新的EBOV抑制剂MBX 2254和MBX 2270，其可以作为开发抗EBOV治疗剂的起点。我们的研究结果还强调了NPC 1-GP相互作用在EBOV进入中的重要性以及NPC 1作为抗丝状病毒治疗靶点的吸引力。
Background. The current Ebola virus (EBOV) outbreak has highlighted the troubling absence of available antivirals or vaccines to treat infected patients and stop the spread of EBOV. The EBOV glycoprotein (GP) plays critical roles in the early stage of virus infection, including receptor binding and membrane fusion, making it a potential target for the development of anti-EBOV drugs. We report the identification of 2 novel EBOV inhibitors targeting viral entry.Methods. To identify small molecule inhibitors of EBOV entry, we carried out a cell-based high-throughput screening using human immunodeficiency virus-based pseudotyped viruses expressing EBOV-GP. Two compounds were identified, and mechanism-of-action studies were performed using immunoflourescence, AlphaLISA, and enzymatic assays for cathepsin B inhibition.Results. We report the identification of 2 novel entry inhibitors. These inhibitors (1) inhibit EBOV infection (50% inhibitory concentration, approximately 0.28 and approximately 10 mu mol/L) at a late stage of entry, (2) induce Niemann-Pick C phenotype, and (3) inhibit GP-Niemann-Pick C1 (NPC1) protein interaction.Conclusions. We have identified 2 novel EBOV inhibitors, MBX2254 and MBX2270, that can serve as starting points for the development of an anti-EBOV therapeutic agent. Our findings also highlight the importance of NPC1-GP interaction in EBOV entry and the attractiveness of NPC1 as an antifiloviral therapeutic target.