title: Merged screening for human immunodeficiency virus Tat and Rev inhibitors.
abstract: In addition to "conventional" drug discovery targets used in modern strategies, mainly focusing on proteins, recent insights into gene regulation as a novel drug concept have begun to invite the targeting of biomolecular interactions between proteins and RNA. Because two protein-RNA interactions (Tat and trans-activation-responsive element, Rev and Rev-responsive element) are essential for any productive replication of human immunodeficiency virus, this important human pathogen was used as a model system for our studies. The design of a fluorescence-based high throughput assay, in which both targets were presented in the same vessel, enabled us to simultaneously interrogate two characteristics of a potential inhibitor: potency of interference and selectivity toward each of the interactions. Although related systems have been reported for several DNA binders, an extension into interference with transcription events would open a new dimension of cellular regulation. Here we describe the setup of the screening assay for over 110,000 compounds as well as a primary characterization of identified hits. The assay's characteristics demonstrate that a microwell-based dual screening system for RNA binders may add a powerful tool to modern drug discovery.
authors.affiliation: Novartis Pharma AG, Basel, Switzerland.
authors.forename: F
authors.initials: F
authors.lastname: Hamy
authors.forename: E
authors.initials: E
authors.lastname: Felder
authors.forename: K
authors.initials: K
authors.lastname: Lipson
authors.forename: T
authors.initials: T
authors.lastname: Klimkait
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015694:Gene Products, rev / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D015696:Gene Products, tat / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006678:HIV / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D006801:Humans; D008904:Miniaturization; D008954:Models, Biological; D018716:Molecular Mimicry; D020871:RNA Stability; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D013696:Temperature; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D015694:Gene Products, rev; D015696:Gene Products, tat; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus