PMID- 27654301 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20191120 IS - 1098-5514 (Electronic) IS - 0022-538X (Print) IS - 0022-538X (Linking) VI - 90 IP - 23 DP - 2016 Dec 1 TI - A Novel Benzodiazepine Compound Inhibits Yellow Fever Virus Infection by Specifically Targeting NS4B Protein. PG - 10774-10788 LID - 10.1128/JVI.01253-16 [doi] AB - Although a highly effective vaccine is available, the number of yellow fever cases has increased over the past 2 decades, which highlights the pressing need for antiviral therapeutics. In a high-throughput screening campaign, we identified an acetic acid benzodiazepine (BDAA) compound which potently inhibits yellow fever virus (YFV). Interestingly, while treatment of YFV-infected cultures with 2 muM BDAA reduced the virion production by greater than 2 logs, the compound was not active against 21 other viruses from 14 different viral families. Selection and genetic analysis of drug-resistant viruses revealed that replacement of the proline at amino acid 219 (P219) of the nonstructural protein 4B (NS4B) with serine, threonine, or alanine conferred YFV with resistance to BDAA without apparent loss of replication fitness in cultured mammalian cells. However, replacement of P219 with glycine conferred BDAA resistance with significant loss of replication ability. Bioinformatics analysis predicts that the P219 amino acid is localized at the endoplasmic reticulum lumen side of the fifth putative transmembrane domain of NS4B, and the mutation may render the viral protein incapable of interacting with BDAA. Our studies thus revealed an important role and the structural basis for the NS4B protein in supporting YFV replication. Moreover, in YFV-infected hamsters, oral administration of BDAA protected 90% of the animals from death, significantly reduced viral load by greater than 2 logs, and attenuated virus infection-induced liver injury and body weight loss. The encouraging preclinical results thus warrant further development of BDAA or its derivatives as antiviral agents to treat yellow fever. IMPORTANCE Yellow fever is an acute viral hemorrhagic disease which threatens approximately 1 billion people living in tropical areas of Africa and Latin America. Although a highly effective yellow fever vaccine has been available for more than 7 decades, the low vaccination rate fails to prevent outbreaks in at-risk regions. It has been estimated that up to 1.7 million YFV infections occur in Africa each year, resulting in 29,000 to 60,000 deaths. Thus far, there is no specific antiviral treatment for yellow fever. To cope with this medical challenge, we identified a benzodiazepine compound that selectively inhibits YFV by targeting the viral NS4B protein. To our knowledge, this is the first report demonstrating in vivo safety and antiviral efficacy of a YFV NS4B inhibitor in an animal model. We have thus reached a critical milestone toward the development of specific antiviral therapeutics for clinical management of yellow fever. CI - Copyright (c) 2016, American Society for Microbiology. All Rights Reserved. FAU - Guo, Fang AU - Guo F AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Wu, Shuo AU - Wu S AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Julander, Justin AU - Julander J AD - Institute for Antiviral Research, Utah State University, Logan, Utah, USA. FAU - Ma, Julia AU - Ma J AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Zhang, Xuexiang AU - Zhang X AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Kulp, John AU - Kulp J AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Cuconati, Andrea AU - Cuconati A AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Block, Timothy M AU - Block TM AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Du, Yanming AU - Du Y AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Guo, Ju-Tao AU - Guo JT AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA. FAU - Chang, Jinhong AU - Chang J AD - Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, Pennsylvania, USA jinhong.chang@bblumberg.org. LA - eng GR - HHSN272201000039C/AI/NIAID NIH HHS/United States GR - HHSN272201100019C/AI/NIAID NIH HHS/United States GR - HHSN272201100022C/AI/NIAID NIH HHS/United States GR - HHSN272201100019I/AI/NIAID NIH HHS/United States GR - HHSN272201000039I/AI/NIAID NIH HHS/United States GR - HHSN272201100022I/AI/NIAID NIH HHS/United States GR - R01 AI104636/AI/NIAID NIH HHS/United States PT - Journal Article DEP - 20161114 PL - United States TA - J Virol JT - Journal of virology JID - 0113724 PMC - PMC5110185 EDAT- 2016/09/23 06:00 MHDA- 2016/09/23 06:01 PMCR- 2017/05/14 CRDT- 2016/09/23 06:00 PHST- 2016/06/27 00:00 [received] PHST- 2016/09/15 00:00 [accepted] PHST- 2016/09/23 06:00 [pubmed] PHST- 2016/09/23 06:01 [medline] PHST- 2016/09/23 06:00 [entrez] PHST- 2017/05/14 00:00 [pmc-release] AID - JVI.01253-16 [pii] AID - 01253-16 [pii] AID - 10.1128/JVI.01253-16 [doi] PST - epublish SO - J Virol. 2016 Nov 14;90(23):10774-10788. doi: 10.1128/JVI.01253-16. Print 2016 Dec 1.