TY - JOUR
T1 - Discovery of Influenza Polymerase PA-PB1 Interaction Inhibitors Using an in Vitro Split-Luciferase Complementation-Based Assay
AU - Zhang, Jiantao
AU - Hu, Yanmei
AU - Wu, Nan
AU - Wang, Jun
N1 - Funding Information:
We thank I. Ghosh for providing pcDNA3.1-Kz-FKBP-27aa-Nfluc and pCDNA3.1-Kz-CFluc-25aa-FRB plasmids, H. Li for providing pGEX-6P-1-GST-Cluc398-NS3 and pET28a-NLuc416-NS2B-E66_stop plasmids, and E. Fodor for providing pcDNA-PA-GFP and pcDNA-PB1 plasmids. This work was supported by the University of Arizona startup fund, the Arizona Biomedical Research Centre Young Investigator grant, and the National Institutes of Health AI 119187 and AI 144887 to J.W.
PY - 2020/1/17
Y1 - 2020/1/17
N2 - The limited therapeutic options and increasing drug-resistance call for next-generation influenza antivirals. Due to the essential function in viral replication and high sequence conservation among influenza viruses, influenza polymerase PA-PB1 protein-protein interaction becomes an attractive drug target. Here, we developed an in vitro split luciferase complementation-based assay to speed up screening of PA-PB1 interaction inhibitors. By screening 10,000 compounds, we identified two PA-PB1 interaction inhibitors, R160792 and R151785, with potent and broad-spectrum antiviral activity against a panel of influenza A and B viruses, including amantadine-, oseltamivir-, or dual resistant strains. Further mechanistic study reveals that R151785 inhibits PA nuclear localization, reduces the levels of viral RNAs and proteins, and inhibits viral replication at the intermediate stage, all of which are in line with its antiviral mechanism of action. Overall, we developed a robust high throughput-screening assay for screening broad-spectrum influenza antivirals targeting PA-PB1 interaction and identified R151785 as a promising antiviral drug candidate.
AB - The limited therapeutic options and increasing drug-resistance call for next-generation influenza antivirals. Due to the essential function in viral replication and high sequence conservation among influenza viruses, influenza polymerase PA-PB1 protein-protein interaction becomes an attractive drug target. Here, we developed an in vitro split luciferase complementation-based assay to speed up screening of PA-PB1 interaction inhibitors. By screening 10,000 compounds, we identified two PA-PB1 interaction inhibitors, R160792 and R151785, with potent and broad-spectrum antiviral activity against a panel of influenza A and B viruses, including amantadine-, oseltamivir-, or dual resistant strains. Further mechanistic study reveals that R151785 inhibits PA nuclear localization, reduces the levels of viral RNAs and proteins, and inhibits viral replication at the intermediate stage, all of which are in line with its antiviral mechanism of action. Overall, we developed a robust high throughput-screening assay for screening broad-spectrum influenza antivirals targeting PA-PB1 interaction and identified R151785 as a promising antiviral drug candidate.
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U2 - 10.1021/acschembio.9b00552
DO - 10.1021/acschembio.9b00552
M3 - Article
C2 - 31714745
AN - SCOPUS:85075685530
VL - 15
SP - 74
EP - 82
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 1
ER -