title: HTS identifies novel and specific uncompetitive inhibitors of the two-component NS2B-NS3 proteinase of West Nile virus.
abstract: West Nile virus (WNV), a member of the Flavividae family, is a mosquito-borne, emerging pathogen. In addition to WNV, the family includes dengue, yellow fever, and Japanese encephalitis viruses, which affect millions of individuals worldwide. Because countermeasures are currently unavailable, flaviviral therapy is urgently required. The flaviviral two-component nonstructural NS2B-NS3 proteinase (protease [pro]) is essential for viral life cycle and, consequently, is a promising drug target. We report here the results of the miniaturization of an NS2B-NS3pro activity assay, followed by high-throughput screening of the National Institutes of Health's 65,000 compound library and identification of novel, uncompetitive inhibitors of WNV NS2B-NS3pro that appear to interfere with the productive interactions of the NS2B cofactor with the NS3pro domain. We anticipate that following structure optimization, the identified probes could form the foundation for the design of novel and specific therapeutics for WNV infection. We also provide the structural basis for additional species-selective allosteric inhibitors of flaviviruses.
authors.affiliation: Pittsburgh Molecular Library Screening Center, Department of Pharmacology, University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA 15260, USA. paj18@pitt.edu
authors.forename: Paul A
authors.initials: PA
authors.lastname: Johnston
authors.forename: Jennifer
authors.initials: J
authors.lastname: Phillips
authors.forename: Tong Ying
authors.initials: TY
authors.lastname: Shun
authors.forename: Sunita
authors.initials: S
authors.lastname: Shinde
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
authors.forename: Donna M
authors.initials: DM
authors.lastname: Huryn
authors.forename: Michael C
authors.initials: MC
authors.lastname: Myers
authors.forename: Boris I
authors.initials: BI
authors.lastname: Ratnikov
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Smith
authors.forename: Ying
authors.initials: Y
authors.lastname: Su
authors.forename: Russell
authors.initials: R
authors.lastname: Dahl
authors.forename: Nicholas D P
authors.initials: ND
authors.lastname: Cosford
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Shiryaev
authors.forename: Alex Y
authors.initials: AY
authors.lastname: Strongin
mesh_terms: D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D002853:Chromatography, Liquid; D003627:Data Interpretation, Statistical; D004353:Drug Evaluation, Preclinical; D004355:Drug Stability; D007202:Indicators and Reagents; D008024:Ligands; D013058:Mass Spectrometry; D008958:Models, Molecular; D011480:Protease Inhibitors / Q000138:chemical synthesis* / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D014901:West Nile Fever / Q000821:virology; D014902:West Nile virus / Q000187:drug effects / Q000201:enzymology*
chemical_list: D000998:Antiviral Agents; D007202:Indicators and Reagents; D008024:Ligands; C080102:NS2B protein, flavivirus; D011480:Protease Inhibitors; D017361:Viral Nonstructural Proteins