TY - JOUR
T1 - Ebselen, a small-molecule capsid inhibitor of HIV-1 replication
AU - Thenin-Houssier, Suzie
AU - De Vera, Ian Mitchelle S.
AU - Pedro-Rosa, Laura
AU - Brady, Angela
AU - Richard, Audrey
AU - Konnick, Briana
AU - Opp, Silvana
AU - Buffone, Cindy
AU - Fuhrmann, Jakob
AU - Kota, Smitha
AU - Billack, Blase
AU - Pietka-Ottlik, Magdalena
AU - Tellinghuisen, Timothy
AU - Choe, Hyeryun
AU - Spicer, Timothy
AU - Scampavia, Louis
AU - Diaz-Griffero, Felipe
AU - Kojetin, Douglas J.
AU - Valentea, Susana T.
N1 - Funding Information:
We are grateful to the NIH AIDS reagent program, Division of AIDS, NIAID, NIH, for the HIV-1 p24 monoclonal antibody (183-H12-5C) from Bruce Chesebro and Kathy Wehrly and for providing all the HIV-1 primary isolates and drug-resistant viruses. Analogs of Ebselen, S-, O-, and C-Ebselen, were a gift from Barbara Slusher, Johns Hopkins University School of Medicine, Baltimore, MD. HeLa-CD4-LTR-LacZ and HeLa-CD4-LTR-Luc cells were provided by Uriel Hazan, Université de Cachan, France. Human hepatocarcinoma cell line Huh7.5 cells were provided by Charles Rice of Rockefeller University. We thank Massimo Caputi for providing the GST-CTD and CTD-Flag constructs. CAI peptide was a gift from Asim K. Debnath. We also honor the memory of our friend and colleague Donny Strosberg, who initiated this project. We declare that we have no conflicts of interest.
Funding Information:
We are grateful to the NIH AIDS reagent program, Division of AIDS, NIAID, NIH, for the HIV-1 p24 monoclonal antibody (183-H12-5C) from Bruce Chesebro and Kathy Wehrly and for providing all the HIV-1 primary isolates and drug-resistant viruses. Analogs of Ebselen, S-, O-, and C-Ebselen, were a gift from Barbara Slusher, Johns Hopkins University School of Medicine, Baltimore, MD. HeLa-CD4-LTR-LacZ and HeLa-CD4-LTR-Luc cells were provided by Uriel Hazan, Universit? de Cachan, France. Human hepatocarcinoma cell line Huh7.5 cells were provided by Charles Rice of Rockefeller University. We thank Massimo Caputi for providing the GST-CTD and CTD-Flag constructs. CAI peptide was a gift from Asim K. Debnath. We also honor the memory of our friend and colleague Donny Strosberg, who initiated this project. We declare that we have no conflicts of interest.
Publisher Copyright:
Copyright © 2016, American Society for Microbiology. All Rights Reserved.
PY - 2016/4
Y1 - 2016/4
N2 - 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 1,280 in 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.
AB - 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 1,280 in 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.
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U2 - 10.1128/AAC.02574-15
DO - 10.1128/AAC.02574-15
M3 - Article
C2 - 26810656
AN - SCOPUS:84963763473
SN - 0066-4804
VL - 60
SP - 2195
EP - 2208
JO - Antimicrobial Agents and Chemotherapy
JF - Antimicrobial Agents and Chemotherapy
IS - 4
ER -
