DISCOVERY OF SELECTIVE, SMALL-MOLECULE INHIBITORS OF RNA COMPLEXES .1. THE TAT PROTEIN TAR RNA COMPLEXES REQUIRED FOR HIV-1 TRANSCRIPTION

We have developed a therapeutic program focusing on the inhibition of a human immunodeficiency virus-1 specific protein-RNA interaction. Thi s program begins with a search for small organic molecules that would interfere with the binding of Tat protein to TAR RNA. The methodologie s chosen to study the HIV-1 Tat-TAR interaction and inhibition include gel mobility shift assays, scintillation proximity assays, filtration assays, and mass spectrometry. These methods helped establish in vitr o high-throughput screening assays which rapidly identified Tat-TAR in hibitors from our corporate compound library. Tat-activated reporter g ene assays were then used to investigate the cellular activities of th e Tat-TAR inhibitors. The cellular activity, selectivity, and toxicity data for select Tat-TAR inhibitors were determined. Evaluation of bot h the cellular data and the Tat-TAR inhibition results led to further testing in anti-HIV-l infection assays. (C) 1997 Elsevier Science Ltd.