Development of antiviral drugs inhibiting fusion between viral envelope and cellular membrane

我们发现流感病毒血凝素（HA）的每个亚型（H1-H16）在糖蛋白HA2亚基的a-螺旋区含有一致的序列。在培养基中加入具有该序列的合成肽后，病毒在MDCK细胞中的复制率降低了10%。由于多肽似乎干扰了细胞中HA的适当折叠，因此使用Chariot将多肽转染到细胞中，战车能够独立于内体途径有效地将多肽输送到培养细胞中。在病毒接种前后给药均无抑制作用。这些数据可能表明，传递肽的内体途径在干扰HA折叠方面非常有效，这是在细胞内囊泡系统中进行的。为了应对未来的流感大流行，我们试图生产针对HA的H5和H2亚型的单克隆抗体。在两种HA亚型的HA球形头部中发现了每个病毒株共有的7个氨基酸残基，并引入到与小鼠I -A^b MHC II类分子结合的框架成分中。用含7个残基的合成肽免疫C57BL/6小鼠脾细胞制备杂交瘤细胞。发现一株细胞系产生抗A/Singapore/1/57 （H2N2）流感病毒的中和抗体。虽然尚未研究抗体与H5病毒的交叉反应性，但预计抗体将为未来流感大流行提供治疗手段。

We found that each subtype (H1-H16) of influenza virus hemagglutinin (HA) contained the consensus sequence in a-helix region of HA2 subunit of the glycoprotein. Virus replication in MDCK cells was reduced by 10% when synthetic peptides with this sequence were added in the medium. Since the peptides seemed to interfere with proper folding of HA in the cells, the peptides were transfected into the cells using Chariot, which is capable of efficiently delivering peptide into cultured cells independently of the endosomal pathway. No inhibition was found by delivering the peptides before or after virus inoculation. These data may indicate that the endosomal pathway for delivering the peptides is much effective in the interference of the folding of HA, which is carried out in the intracellular vesicle system.To prepare for the future pandemic of influenza, we attempted to produce monoclonal antibodies against H5 and H2 subtypes of HA. Seven amino acid residues common to each virus strain tested were found in the globular head of HA of both HA subtypes and introduced into the frame component bound to mouse I -A^b MHC class II molecule. The splenocytes from C57BL/6 mice immunized with the synthetic peptides containing the seven residues were used to prepare hybridoma cells. One cell line was found to produce neutralizing antibodies against A/Singapore/1/57 (H2N2) influenza virus. Although cross-reactivity with H5 virus of the antibodies is not yet studied, it is expected that the antibodies would provide therapeutic means for future pandemic of influenza.