Development of an AlphaScreen-based HIV-1 integrase dimerization assay for discovery of novel allosteric inhibitors

Journal of Biomolecular Screening
Jonas DemeulemeesterFrauke Christ

Abstract

In recent years, HIV-1 integrase (IN) has become an established target in the field of antiretroviral drug discovery. However, its sole clinically approved inhibitor, the integrase strand transfer inhibitor (INSTI) raltegravir, has a surprisingly low genetic barrier for resistance. Furthermore, the only two other integrase inhibitors currently in advanced clinical trials, elvitegravir and dolutegravir, share its mechanism of action and certain resistance pathways. To maintain a range of treatment options, drug discovery efforts are now turning toward allosteric IN inhibitors, which should be devoid of cross-resistance with INSTIs. As IN requires a precise and dynamic equilibrium between several oligomeric species for its activities, the modulation of this equilibrium presents an interesting allosteric target. We report on the development, characterization, and validation of an AlphaScreen-based assay for high-throughput screening for modulators of HIV-1 IN dimerization. Compounds identified as hits in this assay proved to act as allosteric IN inhibitors. Additionally, the assay offers a flexible platform to study IN dimerization.

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Citations

Jul 10, 2013·Asian Pacific Journal of Tropical Biomedicine·Koosha PaydaryKazem Baesi
Jul 16, 2014·Journal of Molecular Graphics & Modelling·Mario A QuevedoWim Dehaen
Mar 29, 2014·Expert Opinion on Therapeutic Patents·Jonas DemeulemeesterFrauke Christ
Sep 26, 2013·Drug Discovery Today. Technologies
Sep 13, 2013·Journal of Medicinal Chemistry·Roberto Di Santo
Jun 30, 2012·The FEBS Journal·Michal MaesAssaf Friedler
Mar 27, 2015·Retrovirology·Belete Ayele DesimmieZeger Debyser
Jul 6, 2014·Current Opinion in Pharmacology·Cristina TintoriMaurizio Botta
Jan 4, 2015·Trends in Biochemical Sciences·Zeger DebyserRik Gijsbers
Jun 14, 2016·Trends in Pharmacological Sciences·Katerina CermakovaZeger Debyser
Jul 12, 2018·PloS One·Caroline WeydertZeger Debyser
Jan 14, 2018·Retrovirology·Jonas DemeulemeesterZeger Debyser
Jul 3, 2020·Journal of Virology·Tomofumi NakamuraHirotomo Nakata
Apr 18, 2018·BioTechniques·John R VeloriaKevin N Dalby
Jul 6, 2021·Journal of Immunological Methods·Elizabeth A ChristianShihua Lin

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Methods Mentioned

BETA
AlphaScreen
PCR
ion exchange chromatography
electrophoresis
Assay
enzyme-linked immunosorbent assay
ELISA

Software Mentioned

AlphaScreen
GraphPad
EnVision
Excel
Prism

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