Effects of non-nucleoside inhibitors of human immunodeficiency virus type 1 in cell-free recombinant reverse transcriptase assays.
We have employed a cell-free human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) assay to study the effects of non-nucleoside inhibitors of RT (NNRTI) by directly monitoring specific HIV DNA products using a HIV-1 genome-derived template and an oligodeoxynucleotide primer. As previously shown by ourselves and others, nucleoside analog triphosphates, e.g. 3'-azido-3'-deoxythymidine triphosphate and 2',3'-dideoxyadenosine triphosphate, could directly inhibit HIV RT RNA-dependent DNA polymerase activity by causing chain termination, as visualized in a RT reaction that yields specific DNA products. In contrast, each of two NNRTIs, nevirapine and delavirdine, directly inhibited RT activity without causing chain termination effects. We also analyzed interactions between nucleoside analogs and NNRTIs or among NNRTIs by chain elongation/dNTP incorporation and/or steady-state kinetic assays. Combinations of nevirapine with the triphosphates of either the (-)-strand of 2',3'-dideoxy-3'-thiacytidine or 2',3'-dideoxyadenosine yielded additive/synergistic effects on RT activity. However, only an additive effect was observed when combinations of nevirapine and 3'-azido-3'-deoxythymidine triphosphate were employed. Combinations of nevirapine and delavirdine had an antagonistic effect on the inhibition of HIV-1 RT activity.
McGill University AIDS Centre, Lady Davis Institute-Jewish General Hospital, Montreal, Quebec, Canada.
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D002474:Cell-Free System; D020008:Delavirdine; D003845:Deoxycytosine Nucleotides / Q000494:pharmacology; D054306:Dideoxynucleotides; D004336:Drug Antagonism; D004357:Drug Synergism; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000187:drug effects*; D007211:Indoles / Q000494:pharmacology; D019829:Nevirapine; D010879:Piperazines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000187:drug effects*; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology*; D013942:Thymine Nucleotides / Q000494:pharmacology; D015215:Zidovudine / Q000031:analogs & derivatives / Q000494:pharmacology
D003845:Deoxycytosine Nucleotides; D054306:Dideoxynucleotides; D007211:Indoles; D010879:Piperazines; D011725:Pyridines; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D015215:Zidovudine; C052873:zidovudine triphosphate; D019829:Nevirapine; D020008:Delavirdine; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase