High-throughput identification of compounds targeting influenza RNA-dependent RNA polymerase activity.
As influenza viruses have developed resistance towards current drugs, new inhibitors that prevent viral replication through different inhibitory mechanisms are useful. In this study, we developed a screening procedure to search for new antiinfluenza inhibitors from 1,200,000 compounds and identified previously reported as well as new antiinfluenza compounds. Several antiinfluenza compounds were inhibitory to the influenza RNA-dependent RNA polymerase (RdRP), including nucleozin and its analogs. The most potent nucleozin analog, 3061 (FA-2), inhibited the replication of the influenza A/WSN/33 (H1N1) virus in MDCK cells at submicromolar concentrations and protected the lethal H1N1 infection of mice. Influenza variants resistant to 3061 (FA-2) were isolated and shown to have the mutation on nucleoprotein (NP) that is distinct from the recently reported resistant mutation of Y289H [Kao R, et al. (2010) Nat Biotechnol 28:600]. Recombinant influenza carrying the Y52H NP is also resistant to 3061 (FA-2), and NP aggregation induced by 3061 (FA-2) was identified as the most likely cause for inhibition. In addition, we identified another antiinfluenza RdRP inhibitor 367 which targets PB1 protein but not NP. A mutant resistant to 367 has H456P mutation at the PB1 protein and both the recombinant influenza and the RdRP expressing the PB1 H456P mutation have elevated resistance to 367. Our high-throughput screening (HTS) campaign thus resulted in the identification of antiinfluenza compounds targeting RdRP activity.
Su
Ching-Yao
CY
Genomics Research Center, Academia Sinica, Taipei.
Cheng
Ting-Jen R
TJ
Lin
Meng-I
MI
Wang
Shi-Yun
SY
Huang
Wen-I
WI
Lin-Chu
Shao-Ying
SY
Chen
Yu-Hou
YH
Wu
Chung-Yi
CY
Lai
Michael M C
MM
Cheng
Wei-Chieh
WC
Wu
Ying-Ta
YT
Tsai
Ming-Daw
MD
Cheng
Yih-Shyun E
YS
Wong
Chi-Huey
CH
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D004285:Dogs; D024882:Drug Resistance, Viral / Q000235:genetics; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects* / Q000201:enzymology; D007251:Influenza, Human / Q000188:drug therapy* / Q000517:prevention & control / Q000821:virology; D051379:Mice; D009698:Nucleoproteins / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors*; D014764:Viral Proteins / Q000037:antagonists & inhibitors; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D009698:Nucleoproteins; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase