title: A Potent Anti-influenza Compound Blocks Fusion through Stabilization of the Prefusion Conformation of the Hemagglutinin Protein.
abstract: An ultrahigh-throughput screen was performed to identify novel small molecule inhibitors of influenza virus replication. The screen employed a recombinant influenza A/WSN/33 virus expressingRenillaluciferase and yielded a hit rate of 0.5%, of which the vast majority showed little cytotoxicity at the inhibitory concentration. One of the top hits from this screen, designated S20, inhibits HA-mediated membrane fusion. S20 shows potent antiviral activity (IC50= 80 nM) and low toxicity (CC50= 40 μM), yielding a selectivity index of 500 and functionality against all of the group 1 influenza A viruses tested in this study, including the pandemic H1N1 and avian H5N1 viruses. Mechanism of action studies proved a direct S20-HA interaction and showed that S20 inhibits fusion by stabilizing the prefusion conformation of HA. In silico docking studies were performed, and the predicted binding site in HA2 corresponds with the area where resistance mutations occurred and correlates with the known role of this region in fusion. This high-throughput screen has yielded many promising new lead compounds, including S20, which will potentially shed light on the molecular mechanisms of viral infection and serve as research tools or be developed for clinical use as antivirals.
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Kris M
authors.initials: KM
authors.lastname: White
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Paul
authors.initials: P
authors.lastname: De Jesus
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, United States.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Pablo
authors.initials: P
authors.lastname: Abreu
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Elisa
authors.initials: E
authors.lastname: Barile
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, United States.
authors.forename: Puiying A
authors.initials: PA
authors.lastname: Mak
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, United States.
authors.forename: Paul
authors.initials: P
authors.lastname: Anderson
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Quy T
authors.initials: QT
authors.lastname: Nguyen
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Inoue
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Silke
authors.initials: S
authors.lastname: Stertz
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Renate
authors.initials: R
authors.lastname: Koenig
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Pellecchia
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Peter
authors.initials: P
authors.lastname: Palese
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, United States.
authors.forename: Kelli
authors.initials: K
authors.lastname: Kuhen
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Adolfo
authors.initials: A
authors.lastname: García-Sastre
authors.affiliation: Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute , 10901 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Sumit K
authors.initials: SK
authors.lastname: Chanda
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.
authors.forename: Megan L
authors.initials: ML
authors.lastname: Shaw
keywords: antiviral; docking; fusion; hemagglutinin; influenza; screen