Nasser E H, Judd A K, Sanchez A, Anastasiou D, Bucher D J
Department of Microbiology and Immunology, New York Medical College, Valhalla 10595, USA.
J Virol. 1996 Dec;70(12):8639-44. doi: 10.1128/JVI.70.12.8639-8644.1996.
Matrix protein (M1) of influenza virus inhibits its own polymerase; this suggested that a peptide segment of M1 with inhibitory properties could serve as an antiviral agent. A peptide synthesized to the Zn2+ finger region of the M1 sequence of influenza virus strain A/PR/8/34 centered around amino acids residues 148 to 166 was shown earlier to be 1,000-fold more effective as a polymerase inhibitor than M1. This peptide, designated peptide 6, represents a Zn2+ finger which includes a 7-residue "loop" and a 4-residue "tail" in addition to the 4 residues on either side of the loop involved in coordination of Zn2+. We have now demonstrated antiviral activity for this peptide in microassays measuring inhibition of the viral cytopathic effect. When the peptide was introduced into tissue culture 5 min after viral challenge with A/PR/8/34, antiviral activity was seen at levels as low as 0.1 nM; on a molar basis, the peptide was shown to be 1,000- to 2,500-fold more effective than ribavirin or amantadine. Antiviral activity was seen with addition of the peptide up to 1 h after viral infection; however, little or no activity was seen at later times, suggesting that viral replication is inhibited at an early stage, possibly at the level of transcription. Reduction in the finger loop or tail length reduced antiviral activity; reduction in the number of residues involved in coordination of Zn2+ abolished antiviral activity. In addition to A/PR/8/34, peptide 6 was shown to have antiviral activity against other type A influenza viruses, including those representing H1N1, H2N2, and H3N2 subtypes. Antiviral activity against type B influenza viruses was also seen. A low level of activity against vesicular stomatitis virus was observed. Zn2+ finger peptides or analogs of Zn2+ finger peptides may provide a new class of antiviral agents effective against influenza virus and possibly other viruses.
流感病毒的基质蛋白（M1）可抑制其自身的聚合酶；这表明具有抑制特性的M1肽段可作为一种抗病毒剂。先前已表明，针对甲型流感病毒A/PR/8/34 M1序列的锌指区域合成的一种肽，以氨基酸残基148至166为中心，作为聚合酶抑制剂的效力比M1高1000倍。这种肽被命名为肽6，它代表一个锌指，除了参与锌离子配位的环两侧的4个残基外，还包括一个7残基的“环”和一个4残基的“尾”。我们现在已经在测量病毒细胞病变效应抑制的微量试验中证明了这种肽的抗病毒活性。当在病毒用A/PR/8/34攻击后5分钟将该肽引入组织培养时，在低至0.1 nM的水平就可观察到抗病毒活性；以摩尔计，该肽比利巴韦林或金刚烷胺有效1000至2500倍。在病毒感染后长达1小时添加该肽时可观察到抗病毒活性；然而，在更晚的时间几乎没有或没有活性，这表明病毒复制在早期被抑制，可能在转录水平。锌指环或尾长度的减少会降低抗病毒活性；参与锌离子配位的残基数量减少会消除抗病毒活性。除了A/PR/8/34外，肽6还显示出对其他甲型流感病毒具有抗病毒活性，包括代表H1N1、H2N2和H3N2亚型的病毒。对乙型流感病毒也观察到了抗病毒活性。观察到对水疱性口炎病毒有低水平的活性。锌指肽或锌指肽类似物可能提供一类新的有效对抗流感病毒及可能其他病毒的抗病毒剂。
