Inhibition of influenza A virus replication by compounds interfering with the fusogenic function of the viral hemagglutinin.
Several compounds that specifically inhibited replication of the H1 and H2 subtypes of influenza virus type A were identified by screening a chemical library for antiviral activity. In single-cycle infections, the compounds inhibited virus-specific protein synthesis when added before or immediately after infection but were ineffective when added 30 min later, suggesting that an uncoating step was blocked. Sequencing of hemagglutinin (HA) genes of several independent mutant viruses resistant to the compounds revealed single amino acid changes that clustered in the stem region of the HA trimer in and near the HA2 fusion peptide. One of the compounds, an N-substituted piperidine, could be docked in a pocket in this region by computer-assisted molecular modeling. This compound blocked the fusogenic activity of HA, as evidenced by its inhibition of low-pH-induced cell-cell fusion in infected cell monolayers. An analog which was more effective than the parent compound in inhibiting virus replication was synthesized. It was also more effective in blocking other manifestations of the low-pH-induced conformational change in HA, including virus inactivation, virus-induced hemolysis of erythrocytes, and susceptibility of the HA to proteolytic degradation. Both compounds inhibited viral protein synthesis and replication more effectively in cells infected with a virus mutated in its M2 protein than with wild-type virus. The possible functional relationship between M2 and HA suggested by these results is discussed.
Plotch
S J
SJ
Department of Molecular Biology, Infectious Disease Section, Wyeth-Ayerst Research, Pearl River, New York 10965, USA. Plotchs@war.wyeth.com
O'Hara
B
B
Morin
J
J
Palant
O
O
LaRocque
J
J
Bloom
J D
JD
Lang
S A
SA
DiGrandi
M J
MJ
Bradley
M
M
Nilakantan
R
R
Gluzman
Y
Y
D000547:Amantadine / Q000494:pharmacology; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002459:Cell Fusion / Q000187:drug effects*; D002460:Cell Line; D003001:Cloning, Molecular; D003198:Computer Simulation; D006389:Hemagglutinins, Viral / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000187:drug effects* / Q000502:physiology; D008958:Models, Molecular; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D006389:Hemagglutinins, Viral; D011500:Protein Synthesis Inhibitors; D014764:Viral Proteins; D000547:Amantadine