Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection

Jagannadha K. Sastry, Pramod N. Nehete, Saeed Khan, Billie J. Nowak, William Plunkett, Ralph B. Arlinghaus, David Farquhar

Research output: Contribution to journalArticle

Abstract

2′,3′-Dideoxyuridine (ddU) is ineffective at controlling human immunodeficiency virus type 1 (HIV-1) infection in human T cells, because it is not biotransformed to the active 5′-triphosphate. The metabolic block resides in the poor substrate affinity of ddU for cellular nucleoside kinases. This problem cannot be overcome by supplying the preformed nucleotides, because such compounds are unable to penetrate cells. To circumvent the requirement of ddU for enzymic phosphorylation, we have prepared bis(pivaloyloxymethyl) 2′,3′-dideoxyuridine 5′-monophosphate (piv2 ddUMP), as a potential membrane-permeable prodrug of ddUMP, and investigated its metabolism and anti-HIV activity in two human T cell lines, one with wild-type thymidine kinase activity (MT-4) and the other deficient in thymidine kinase activity (CEM-tk-). The 5′-mono-, di-, and triphosphates of ddU were formed in both cell lines after exposure to piv2-ddUMP. In contrast, phosphorylated metabolites were not observed in cells treated with ddU or ddUMP alone. piv2-ddUMP also reduced the cytopathic effects of HIV-1 in MT-4 cells (ED50, 4.75 μM) and inhibited virus production in culture fluid (ED50, 20 μM). In addition, piv2-ddUMP protected CEM-tk- cells from HIV-1 infection, as demonstrated by inhibition of intracellular p24 antigen levels (ED50, 3 μM) and reverse transcriptase activity in culture medium (ED50, 2.5 μM). Based on these findings, we propose that the "masked nucleotide" strategy may make available for development nucleoside analogues hitherto considered inactive because of failure to undergo biotransformation to the corresponding 5′-monophosphates. Moreover, by circumventing metabolic dependency on nucleoside kinases, the strategy may overcome acquired resistance to nucleoside analogues caused by the loss or depletion of nucleoside kinases.

Original languageEnglish (US)
Pages (from-to)441-445
Number of pages5
JournalMolecular Pharmacology
Volume41
Issue number3
StatePublished - Mar 1992
Externally publishedYes

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nucleoside phosphotransferase
Virus Diseases
HIV
HIV-1
Membranes
Thymidine Kinase
Nucleosides
Nucleotides
T-Lymphocytes
Cell Line
Diphosphates
RNA-Directed DNA Polymerase
Prodrugs
Biotransformation
Membrane Potentials
Culture Media
Phosphorylation
Viruses
Antigens
bis(pivaloyloxymethyl) 2',3'dideoxyuridine 5'-monophosphate

ASJC Scopus subject areas

  • Pharmacology

Cite this

Sastry, J. K., Nehete, P. N., Khan, S., Nowak, B. J., Plunkett, W., Arlinghaus, R. B., & Farquhar, D. (1992). Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection. Molecular Pharmacology, 41(3), 441-445.

Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection. / Sastry, Jagannadha K.; Nehete, Pramod N.; Khan, Saeed; Nowak, Billie J.; Plunkett, William; Arlinghaus, Ralph B.; Farquhar, David.

In: Molecular Pharmacology, Vol. 41, No. 3, 03.1992, p. 441-445.

Research output: Contribution to journalArticle

Sastry, JK, Nehete, PN, Khan, S, Nowak, BJ, Plunkett, W, Arlinghaus, RB & Farquhar, D 1992, 'Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection', Molecular Pharmacology, vol. 41, no. 3, pp. 441-445.
Sastry JK, Nehete PN, Khan S, Nowak BJ, Plunkett W, Arlinghaus RB et al. Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection. Molecular Pharmacology. 1992 Mar;41(3):441-445.
Sastry, Jagannadha K. ; Nehete, Pramod N. ; Khan, Saeed ; Nowak, Billie J. ; Plunkett, William ; Arlinghaus, Ralph B. ; Farquhar, David. / Membrane-permeable dideoxyuridine 5′-monophosphate analogue inhibits human immunodeficiency virus infection. In: Molecular Pharmacology. 1992 ; Vol. 41, No. 3. pp. 441-445.
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AU - Sastry, Jagannadha K.

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N2 - 2′,3′-Dideoxyuridine (ddU) is ineffective at controlling human immunodeficiency virus type 1 (HIV-1) infection in human T cells, because it is not biotransformed to the active 5′-triphosphate. The metabolic block resides in the poor substrate affinity of ddU for cellular nucleoside kinases. This problem cannot be overcome by supplying the preformed nucleotides, because such compounds are unable to penetrate cells. To circumvent the requirement of ddU for enzymic phosphorylation, we have prepared bis(pivaloyloxymethyl) 2′,3′-dideoxyuridine 5′-monophosphate (piv2 ddUMP), as a potential membrane-permeable prodrug of ddUMP, and investigated its metabolism and anti-HIV activity in two human T cell lines, one with wild-type thymidine kinase activity (MT-4) and the other deficient in thymidine kinase activity (CEM-tk-). The 5′-mono-, di-, and triphosphates of ddU were formed in both cell lines after exposure to piv2-ddUMP. In contrast, phosphorylated metabolites were not observed in cells treated with ddU or ddUMP alone. piv2-ddUMP also reduced the cytopathic effects of HIV-1 in MT-4 cells (ED50, 4.75 μM) and inhibited virus production in culture fluid (ED50, 20 μM). In addition, piv2-ddUMP protected CEM-tk- cells from HIV-1 infection, as demonstrated by inhibition of intracellular p24 antigen levels (ED50, 3 μM) and reverse transcriptase activity in culture medium (ED50, 2.5 μM). Based on these findings, we propose that the "masked nucleotide" strategy may make available for development nucleoside analogues hitherto considered inactive because of failure to undergo biotransformation to the corresponding 5′-monophosphates. Moreover, by circumventing metabolic dependency on nucleoside kinases, the strategy may overcome acquired resistance to nucleoside analogues caused by the loss or depletion of nucleoside kinases.

AB - 2′,3′-Dideoxyuridine (ddU) is ineffective at controlling human immunodeficiency virus type 1 (HIV-1) infection in human T cells, because it is not biotransformed to the active 5′-triphosphate. The metabolic block resides in the poor substrate affinity of ddU for cellular nucleoside kinases. This problem cannot be overcome by supplying the preformed nucleotides, because such compounds are unable to penetrate cells. To circumvent the requirement of ddU for enzymic phosphorylation, we have prepared bis(pivaloyloxymethyl) 2′,3′-dideoxyuridine 5′-monophosphate (piv2 ddUMP), as a potential membrane-permeable prodrug of ddUMP, and investigated its metabolism and anti-HIV activity in two human T cell lines, one with wild-type thymidine kinase activity (MT-4) and the other deficient in thymidine kinase activity (CEM-tk-). The 5′-mono-, di-, and triphosphates of ddU were formed in both cell lines after exposure to piv2-ddUMP. In contrast, phosphorylated metabolites were not observed in cells treated with ddU or ddUMP alone. piv2-ddUMP also reduced the cytopathic effects of HIV-1 in MT-4 cells (ED50, 4.75 μM) and inhibited virus production in culture fluid (ED50, 20 μM). In addition, piv2-ddUMP protected CEM-tk- cells from HIV-1 infection, as demonstrated by inhibition of intracellular p24 antigen levels (ED50, 3 μM) and reverse transcriptase activity in culture medium (ED50, 2.5 μM). Based on these findings, we propose that the "masked nucleotide" strategy may make available for development nucleoside analogues hitherto considered inactive because of failure to undergo biotransformation to the corresponding 5′-monophosphates. Moreover, by circumventing metabolic dependency on nucleoside kinases, the strategy may overcome acquired resistance to nucleoside analogues caused by the loss or depletion of nucleoside kinases.

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