Discovery of selective, small-molecule inhibitors of RNA complexes--I. 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. 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.
Mei
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HY
BioOrganic Chemistry Section, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI 48106, USA.
Mack
D P
DP
Galan
A A
AA
Halim
N S
NS
Heldsinger
A
A
Loo
J A
JA
Moreland
D W
DW
Sannes-Lowery
K A
KA
Sharmeen
L
L
Truong
H N
HN
Czarnik
A W
AW
D000595:Amino Acid Sequence; D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001483:Base Sequence; D001570:Benzodiazepinones / Q000494:pharmacology; D002166:Camptothecin / Q000494:pharmacology; D015696:Gene Products, tat / Q000187:drug effects* / Q000378:metabolism*; D015497:HIV-1 / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D011758:Pyrroles / Q000494:pharmacology; D012367:RNA, Viral / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*; D015533:Transcriptional Activation / Q000187:drug effects / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology; D054322:tat Gene Products, Human Immunodeficiency Virus
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D019380:Anti-HIV Agents; D000972:Antineoplastic Agents, Phytogenic; D001570:Benzodiazepinones; D015696:Gene Products, tat; D011758:Pyrroles; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus; C072173:Ro 5-3335; D002166:Camptothecin