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Characterization of Potent Fusion Inhibitors of Influenza Virus

New inhibitors of influenza viruses are needed to combat the potential emergence of novel human influenza viruses. We have identified a class of small molecules that inhibit replication of influenza virus at picomolar concentrations in plaque reduction assays. The compound also inhibits replication...

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Autores principales: Rowse, Michael, Qiu, Shihong, Tsao, Jun, Xian, Tongmei, Khawaja, Sarah, Yamauchi, Yohei, Yang, Zhen, Wang, Guoxin, Luo, Ming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4372562/
https://www.ncbi.nlm.nih.gov/pubmed/25803288
http://dx.doi.org/10.1371/journal.pone.0122536
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author Rowse, Michael
Qiu, Shihong
Tsao, Jun
Xian, Tongmei
Khawaja, Sarah
Yamauchi, Yohei
Yang, Zhen
Wang, Guoxin
Luo, Ming
author_facet Rowse, Michael
Qiu, Shihong
Tsao, Jun
Xian, Tongmei
Khawaja, Sarah
Yamauchi, Yohei
Yang, Zhen
Wang, Guoxin
Luo, Ming
author_sort Rowse, Michael
collection PubMed
description New inhibitors of influenza viruses are needed to combat the potential emergence of novel human influenza viruses. We have identified a class of small molecules that inhibit replication of influenza virus at picomolar concentrations in plaque reduction assays. The compound also inhibits replication of vesicular stomatitis virus. Time of addition and dilution experiments with influenza virus indicated that an early time point of infection was blocked and that inhibitor 136 tightly bound to virions. Using fluorescently labeled influenza virus, inhibition of viral fusion to cellular membranes by blocked lipid mixing was established as the mechanism of action for this class of inhibitors. Stabilization of the neutral pH form of hemagglutinin (HA) was ruled out by trypsin digestion studies in vitro and with conformation specific HA antibodies within cells. Direct visualization of 136 treated influenza virions at pH 7.5 or acidified to pH 5.0 showed that virions remain intact and that glycoproteins become disorganized as expected when HA undergoes a conformational change. This suggests that exposure of the fusion peptide at low pH is not inhibited but lipid mixing is inhibited, a different mechanism than previously reported fusion inhibitors. We hypothesize that this new class of inhibitors intercalate into the virus envelope altering the structure of the viral envelope required for fusion to cellular membranes.
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spelling pubmed-43725622015-04-04 Characterization of Potent Fusion Inhibitors of Influenza Virus Rowse, Michael Qiu, Shihong Tsao, Jun Xian, Tongmei Khawaja, Sarah Yamauchi, Yohei Yang, Zhen Wang, Guoxin Luo, Ming PLoS One Research Article New inhibitors of influenza viruses are needed to combat the potential emergence of novel human influenza viruses. We have identified a class of small molecules that inhibit replication of influenza virus at picomolar concentrations in plaque reduction assays. The compound also inhibits replication of vesicular stomatitis virus. Time of addition and dilution experiments with influenza virus indicated that an early time point of infection was blocked and that inhibitor 136 tightly bound to virions. Using fluorescently labeled influenza virus, inhibition of viral fusion to cellular membranes by blocked lipid mixing was established as the mechanism of action for this class of inhibitors. Stabilization of the neutral pH form of hemagglutinin (HA) was ruled out by trypsin digestion studies in vitro and with conformation specific HA antibodies within cells. Direct visualization of 136 treated influenza virions at pH 7.5 or acidified to pH 5.0 showed that virions remain intact and that glycoproteins become disorganized as expected when HA undergoes a conformational change. This suggests that exposure of the fusion peptide at low pH is not inhibited but lipid mixing is inhibited, a different mechanism than previously reported fusion inhibitors. We hypothesize that this new class of inhibitors intercalate into the virus envelope altering the structure of the viral envelope required for fusion to cellular membranes. Public Library of Science 2015-03-24 /pmc/articles/PMC4372562/ /pubmed/25803288 http://dx.doi.org/10.1371/journal.pone.0122536 Text en © 2015 Rowse et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Rowse, Michael
Qiu, Shihong
Tsao, Jun
Xian, Tongmei
Khawaja, Sarah
Yamauchi, Yohei
Yang, Zhen
Wang, Guoxin
Luo, Ming
Characterization of Potent Fusion Inhibitors of Influenza Virus
title Characterization of Potent Fusion Inhibitors of Influenza Virus
title_full Characterization of Potent Fusion Inhibitors of Influenza Virus
title_fullStr Characterization of Potent Fusion Inhibitors of Influenza Virus
title_full_unstemmed Characterization of Potent Fusion Inhibitors of Influenza Virus
title_short Characterization of Potent Fusion Inhibitors of Influenza Virus
title_sort characterization of potent fusion inhibitors of influenza virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4372562/
https://www.ncbi.nlm.nih.gov/pubmed/25803288
http://dx.doi.org/10.1371/journal.pone.0122536
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