title: Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication.
abstract: The human immunodeficiency virus type 1 (HIV-1) capsid plays crucial roles in HIV-1 replication and thus represents an excellent drug target. We developed a high-throughput screening method based on a time-resolved fluorescence resonance energy transfer (HTS-TR-FRET) assay, using the C-terminal domain (CTD) of HIV-1 capsid to identify inhibitors of capsid dimerization. This assay was used to screen a library of pharmacologically active compounds, composed of 1,280in vivo-active drugs, and identified ebselen [2-phenyl-1,2-benzisoselenazol-3(2H)-one], an organoselenium compound, as an inhibitor of HIV-1 capsid CTD dimerization. Nuclear magnetic resonance (NMR) spectroscopic analysis confirmed the direct interaction of ebselen with the HIV-1 capsid CTD and dimer dissociation when ebselen is in 2-fold molar excess. Electrospray ionization mass spectrometry revealed that ebselen covalently binds the HIV-1 capsid CTD, likely via a selenylsulfide linkage with Cys198 and Cys218. This compound presents anti-HIV activity in single and multiple rounds of infection in permissive cell lines as well as in primary peripheral blood mononuclear cells. Ebselen inhibits early viral postentry events of the HIV-1 life cycle by impairing the incoming capsid uncoating process. This compound also blocks infection of other retroviruses, such as Moloney murine leukemia virus and simian immunodeficiency virus, but displays no inhibitory activity against hepatitis C and influenza viruses. This study reports the use of TR-FRET screening to successfully identify a novel capsid inhibitor, ebselen, validating HIV-1 capsid as a promising target for drug development.
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Suzie
authors.initials: S
authors.lastname: Thenin-Houssier
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Ian Mitchelle S
authors.initials: IM
authors.lastname: de Vera
authors.affiliation: Department of Molecular Therapeutics, Molecular Screening Center, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Pedro-Rosa
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Brady
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Audrey
authors.initials: A
authors.lastname: Richard
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Briana
authors.initials: B
authors.lastname: Konnick
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USA.
authors.forename: Silvana
authors.initials: S
authors.lastname: Opp
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USA.
authors.forename: Cindy
authors.initials: C
authors.lastname: Buffone
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Jakob
authors.initials: J
authors.lastname: Fuhrmann
authors.affiliation: Department of Cancer Biology, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Smitha
authors.initials: S
authors.lastname: Kota
authors.affiliation: Department of Pharmaceutical Sciences, St. John's University, Jamaica, New York, USA.
authors.forename: Blase
authors.initials: B
authors.lastname: Billack
authors.affiliation: Department of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Pietka-Ottlik
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Tellinghuisen
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Hyeryun
authors.initials: H
authors.lastname: Choe
authors.affiliation: Department of Molecular Therapeutics, Molecular Screening Center, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Spicer
authors.affiliation: Department of Molecular Therapeutics, Molecular Screening Center, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Scampavia
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USA.
authors.forename: Felipe
authors.initials: F
authors.lastname: Diaz-Griffero
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida, USA.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Kojetin
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, Jupiter, Florida, USA svalente@scripps.edu.
authors.forename: Susana T
authors.initials: ST
authors.lastname: Valente
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry / Q000494:pharmacology*; D001393:Azoles / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites; D002213:Capsid / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D036022:Capsid Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D062313:Databases, Pharmaceutical; D031541:Fluorescence Resonance Energy Transfer; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D054833:Isoindoles; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000821:virology; D008979:Moloney murine leukemia virus / Q000187:drug effects / Q000502:physiology; D016566:Organoselenium Compounds / Q000737:chemistry / Q000494:pharmacology*; D011485:Protein Binding; D000072417:Protein Domains; D055503:Protein Multimerization / Q000187:drug effects; D017433:Protein Structure, Secondary; D015302:Simian Immunodeficiency Virus / Q000187:drug effects / Q000502:physiology; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D019065:Virus Assembly / Q000187:drug effects / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents; D001393:Azoles; D036022:Capsid Proteins; D054833:Isoindoles; D016566:Organoselenium Compounds; D054852:Small Molecule Libraries; C042986:ebselen
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