Design of structure-based reverse transcriptase inhibitors.
Based on the crystallographic structure of the active site in the reverse transcriptase (RT) of human immunodeficiency virus (HIV), a group of hydrophobic polyadenylic acid (5') derivatives were designed and synthesized as inhibitors of the enzyme. These compounds were found to inhibit all six of the RTs tested, with IC50 = 10(-11)-10(-8) M, but did not inhibit either RNA polymerase II (even at 10(-5) M) or DNA polymerase I up to 10(-6) M inhibitor concentration. The underivatized poly(A) did not inhibit any of the RTs tested under the same conditions. In aqueous solutions of purified HIV-1 RT, poly-2'-O-(2,4-dinitrophenyl)-oligo(A) was found to inhibit the enzyme reversibly and compete with the primer-template poly(A)-(dT)12, whereas poly-2'-O-(3-fluoro-4,6-dinitrophenyl)-poly(A) was found to inactivate HIV-1 RT irreversibly by covalent labeling. A comparison of physicochemical properties of the hybrids poly(A)-poly(dT) and dinitrophenyl-poly(A)-poly(dT) shows that the hydrophobic dinitrophenyl groups stabilize double helical structures. These inhibitors were also found to be effective in keeping susceptible lymphocytes viable in the presence of HIV-1 (wild type). The effective inhibitor concentrations (EC50) were found to be 0.2-2.6 microgram/ml. No toxic effect on the host cells was found even at 100-1000-fold higher inhibitor concentrations.
Kang
I
I
Department of Chemistry, State University of New York, Buffalo 14214-3094.
Wang
J H
JH
D000818:Animals; D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D001665:Binding Sites; D002417:Cattle; D018360:Crystallography, X-Ray; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors; D015195:Drug Design; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D009052:Leukemia Virus, Murine / Q000201:enzymology; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000382:microbiology; D008322:Mammals; D011061:Poly A / Q000138:chemical synthesis / Q000494:pharmacology*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry; D018894:Reverse Transcriptase Inhibitors; D013816:Thermodynamics; D013950:Thymus Gland / Q000201:enzymology
D007202:Indicators and Reagents; D018894:Reverse Transcriptase Inhibitors; D011061:Poly A; D012319:RNA Polymerase II; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I