Abstract
We have developed a therapeutic program focusing on the inhibition of a human immunodeficiency virus-1 specific protein-RNA interaction. This program begins with a search for small organic molecules that would interfere with the binding of Tat protein to TAR RNA. The methodologies 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 vitro high-throughput screening assays which rapidly identified Tat-TAR inhibitors from our corporate compound library. Tat-activated reporter gene assays were then used to investigate the cellular activities of the Tat-TAR inhibitors. The cellular activity, selectivity, and toxicity data for select Tat-TAR inhibitors were determined. Evaluation of both the cellular data and the Tat-TAR inhibition results led to further testing in anti-HIV-1 infection assays.
MeSH terms
-
Amino Acid Sequence
-
Aminoglycosides
-
Anti-Bacterial Agents / pharmacology
-
Anti-HIV Agents / pharmacology*
-
Antineoplastic Agents, Phytogenic / pharmacology
-
Base Sequence
-
Benzodiazepinones / pharmacology
-
Camptothecin / pharmacology
-
Gene Products, tat / drug effects*
-
Gene Products, tat / metabolism*
-
HIV-1 / drug effects*
-
HIV-1 / metabolism
-
HIV-1 / physiology*
-
HeLa Cells / drug effects
-
Humans
-
Molecular Sequence Data
-
Pyrroles / pharmacology
-
RNA, Viral / antagonists & inhibitors*
-
RNA, Viral / metabolism*
-
Transcription, Genetic / drug effects*
-
Transcriptional Activation / drug effects
-
Transcriptional Activation / physiology
-
Virus Replication / drug effects
-
Virus Replication / physiology
-
tat Gene Products, Human Immunodeficiency Virus
Substances
-
Aminoglycosides
-
Anti-Bacterial Agents
-
Anti-HIV Agents
-
Antineoplastic Agents, Phytogenic
-
Benzodiazepinones
-
Gene Products, tat
-
Pyrroles
-
RNA, Viral
-
tat Gene Products, Human Immunodeficiency Virus
-
Ro 5-3335
-
Camptothecin