title: Development of an AlphaScreen-based HIV-1 integrase dimerization assay for discovery of novel allosteric inhibitors.
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.
authors.affiliation: Katholieke Universiteit Leuven, Leuven, Belgium.
authors.forename: Jonas
authors.initials: J
authors.lastname: Demeulemeester
authors.forename: Cristina
authors.initials: C
authors.lastname: Tintori
authors.forename: Maurizio
authors.initials: M
authors.lastname: Botta
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
authors.forename: Frauke
authors.initials: F
authors.lastname: Christ
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D057075:Enzyme Assays / Q000379:methods*; D019427:HIV Integrase / Q000737:chemistry* / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D055503:Protein Multimerization / Q000187:drug effects; D011786:Quality Control; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D019428:HIV Integrase Inhibitors; D011993:Recombinant Fusion Proteins; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1