Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs.
Carbovir (the carbocyclic analog of 2'-3'-didehydro-2',3'-dideoxyguanosine) is a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication. Assays were developed to assess the mechanism of inhibition by the 5'-triphosphate of carbovir of HIV-1 reverse transcriptase using either RNA or DNA templates that contain all four natural nucleotides. Carbovir-TP was a potent inhibitor of HIV-1 reverse transcriptase using either template with Ki values similar to that observed by AZT-TP, ddGTP, and ddTTP. The kinetic constants for incorporation of these nucleotide analogs into DNA by HIV-1 reverse transcriptase using either template were similar to the values seen for their respective natural nucleotides. In addition, the incorporation of either carbovir-TP or AZT-TP in the presence of dGTP or dTTP, respectively, indicated that the mechanism of inhibition by these two nucleotide analogs was due to their incorporation into the DNA resulting in chain termination. Carbovir-TP was not a potent inhibitor of DNA polymerase alpha, beta, or gamma, or DNA primase. Given the potent activity of carbovir-TP against HIV-1 reverse transcriptase and its lack of activity against human DNA polymerases, we believe that further evaluation of this compound as a potential drug for the treatment of HIV-1 infection is warranted.
Parker
W B
WB
Kettering-Meyer Laboratory, Southern Research Institute, Birmingham, Alabama 35205.
White
E L
EL
Shaddix
S C
SC
Ross
L J
LJ
Buckheit
R W
RW
Germany
J M
JM
Secrist
J A
JA
Vince
R
R
Shannon
W M
WM
D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis / Q000378:metabolism; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors*; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors*; D004258:DNA Polymerase III / Q000037:antagonists & inhibitors*; D003848:Deoxyguanine Nucleotides / Q000378:metabolism / Q000494:pharmacology*; D054306:Dideoxynucleotides; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008969:Molecular Sequence Data; D012313:RNA / Q000378:metabolism; D018894:Reverse Transcriptase Inhibitors; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism / Q000494:pharmacology; D015215:Zidovudine / Q000031:analogs & derivatives / Q000494:pharmacology; D006160:Guanosine Triphosphate
D000998:Antiviral Agents; D004247:DNA; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004258:DNA Polymerase III; D003848:Deoxyguanine Nucleotides; D054306:Dideoxynucleotides; D012313:RNA; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D015215:Zidovudine; C066928:carbovir triphosphate; D006160:Guanosine Triphosphate; C031709:2',3'-dideoxyguanosine 5'-triphosphate; C052873:zidovudine triphosphate; C029603:deoxyguanosine triphosphate; C017216:2',3'-dideoxythymidine triphosphate; C024157:thymidine 5'-triphosphate