title: Chemical library screens targeting an HIV-1 accessory factor/host cell kinase complex identify novel antiretroviral compounds.
abstract: Nef is an HIV-1 accessory protein essential for AIDS progression and an attractive target for drug discovery. Lack of a catalytic function makes Nef difficult to assay in chemical library screens. We developed a high-throughput screening assay for inhibitors of Nef function by coupling it to one of its host cell binding partners, the Src-family kinase Hck. Hck activation is dependent upon Nef in this assay, providing a direct readout of Nef activity in vitro. Using this screen, a unique diphenylfuropyrimidine was identified as a strong inhibitor of Nef-dependent Hck activation. This compound also exhibited remarkable antiretroviral effects, blocking Nef-dependent HIV replication in cell culture. Structurally related analogs were synthesized and shown to exhibit similar Nef-dependent antiviral activity, identifying the diphenylfuropyrimidine substructure as a new lead for antiretroviral drug development. This study demonstrates that coupling noncatalytic HIV accessory factors with host cell target proteins addressable by high-throughput assays may afford new avenues for the discovery of anti-HIV agents.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Lori
authors.initials: L
authors.lastname: Emert-Sedlak
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kodama
authors.forename: Edwina C
authors.initials: EC
authors.lastname: Lerner
authors.forename: Weixiang
authors.initials: W
authors.lastname: Dai
authors.forename: Caleb
authors.initials: C
authors.lastname: Foster
authors.forename: Billy W
authors.initials: BW
authors.lastname: Day
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Smithgall
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology*; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D004789:Enzyme Activation; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D057166:High-Throughput Screening Assays; D006801:Humans; D015394:Molecular Structure; D011485:Protein Binding; D051572:Proto-Oncogene Proteins c-hck / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014779:Virus Replication; D054311:nef Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
chemical_list: D019380:Anti-HIV Agents; D054852:Small Molecule Libraries; D054311:nef Gene Products, Human Immunodeficiency Virus; C514837:nef protein, Human immunodeficiency virus 1; D051572:Proto-Oncogene Proteins c-hck