Luo G, Colonno R, Krystal M
Department of Virology, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, Connecticut 06492, USA.
Virology. 1996 Dec 1;226(1):66-76. doi: 10.1006/viro.1996.0628.
A novel inhibitor of influenza virus growth in tissue culture was identified and characterized. This compound (BMY-27709) has an IC50 of 3-8 microM for A/WSN/33 virus growth in a multicycle replication assay and is active against all H1 and H2 subtype viruses tested. However, BMY-27709 was found to be inactive against H3 subtype viruses, as well as influenza B/Lee/40 virus. BMY-27709 was also found to act against H1 and H2 viruses early in infection, suggesting that the target for inhibition is the hemagglutinin protein. This was confirmed through the use of reassortant viruses and the isolation of a virus resistant to BMY-27709. The resistant virus isolated contained a phenylalanine to serine change at amino acid 110 of the HA2 subunit. That this single mutation was responsible for the acquisition of resistance to BMY-27709 was proven through reverse genetics, as transfectant virus containing only this change was shown to be resistant to BMY-27709, while the control virus without this mutation remained sensitive. BMY-27709 is able to inhibit virus-induced red blood cell hemolysis, suggesting that it blocks the membrane fusion function of hemagglutinin. These experiments further illustrate that the hemagglutinin protein of influenza virus is a viable target for the discovery and development of small molecule inhibitors of virus growth.
一种新型的流感病毒在组织培养中生长的抑制剂被鉴定和表征。这种化合物（BMY-27709）在多轮复制试验中对A/WSN/33病毒生长的IC50为3-8微摩尔，并且对所有测试的H1和H2亚型病毒均有活性。然而，发现BMY-27709对H3亚型病毒以及B/Lee/40流感病毒无活性。还发现BMY-27709在感染早期对H1和H2病毒起作用，这表明抑制的靶点是血凝素蛋白。通过使用重配病毒和分离对BMY-27709耐药的病毒证实了这一点。分离出的耐药病毒在HA2亚基的第110位氨基酸处发生了苯丙氨酸到丝氨酸的变化。通过反向遗传学证明，仅含有这种变化的转染病毒对BMY-27709耐药，而没有这种突变的对照病毒仍然敏感，从而证明了这种单一突变导致了对BMY-27709的耐药性。BMY-27709能够抑制病毒诱导的红细胞溶血，这表明它阻断了血凝素的膜融合功能。这些实验进一步说明，流感病毒的血凝素蛋白是发现和开发病毒生长小分子抑制剂的可行靶点。
