title: Inhibition of human immunodeficiency virus type 1 reverse transcriptase, RNase H, and integrase activities by hydroxytropolones.
abstract: Human immunodeficiency virus type I reverse transcriptase (RT) possesses distinct DNA polymerase and RNase H sites, whereas integrase (IN) uses the same active site to perform 3'-end processing and strand transfer of the proviral DNA. These four enzymatic activities are essential for viral replication and require metal ions. Two Mg2+ ions are present in the RT polymerase site, and one or two Mg2+ ions are required for the catalytic activities of RNase H and IN. We tested the possibility of inhibition of the RT polymerase and RNase H as well as the IN 3'-end processing and transfer activities of purified enzymes by a series of 3,7-dihydroxytropolones designed to target two Mg2+ ions separated by approximately 3.7 angstroms. The RT polymerase and IN 3' processing and strand transfer activities were inhibited at submicromolar concentrations, while the RNase H activity was inhibited in the low micromolar range. In all cases, the lack of inhibition by tropolones and O-methylated 3,7-dihydroxytropolones was consistent with the active molecules binding the metal ions in the active site. In addition, inhibition of the DNA polymerase activity was shown to depend on the Mg2+ concentration. Furthermore, selective inhibitors were identified for several of the activities tested, leaving some potential for design of improved inhibitors. However, all tested compounds exhibited cellular toxicity that presently limits their applications.
authors.affiliation: Unité Propre de Recherche 9002 du CNRS conventionnée à l'Université Louis Pasteur, IBMC, 15 rue René Descartes, 67084 Strasbourg cedex, France.
authors.forename: Joël
authors.initials: J
authors.lastname: Didierjean
authors.forename: Catherine
authors.initials: C
authors.lastname: Isel
authors.forename: Flore
authors.initials: F
authors.lastname: Querré
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouscadet
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Aubertin
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Valnot
authors.forename: Serge R
authors.initials: SR
authors.lastname: Piettre
authors.forename: Roland
authors.initials: R
authors.lastname: Marquet
mesh_terms: D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019427:HIV Integrase / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D016914:Ribonuclease H / Q000037:antagonists & inhibitors*; D014334:Tropolone / Q000031:analogs & derivatives / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D019428:HIV Integrase Inhibitors; D014334:Tropolone; D019427:HIV Integrase; D054303:HIV Reverse Transcriptase; D016914:Ribonuclease H