Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate

Efficient intracellular delivery of d4TMP

Jan Balzarini, Stefano Aquaro, Tina Knispel, Chiara Rampazzo, Vera Bianchi, Carlo Federico Perno, Erik De Clercq, Chris Meier

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine-5'-monophosphate (cycloSal-d4TMP) is a potent and selective inhibitor of human immunodeficiency virus replication in cell culture and differs from other nucleotide prodrug approaches in that it is designed to selectively deliver the nucleotide 5'-monophosphate by a controlled, chemically induced hydrolysis. Its antiviral efficacy in cell culture is at least as good as, if not superior to, that of d4T. CycloSal-d4TMP was found to lead to the efficient intracellular release of d4TMP in a variety of cell lines, including both wild-type CEM and thymidine kinase-deficient CEM/TK- cells. Under similar experimental conditions, exposure of CEM/TK- cells to d4T failed to result in significant d4TTP levels. The intracellular conversion of cycloSal-d4TMP proved to be both time and dose dependent. The half-life of d4TTP generated intracellularly from d4T- or cycloSal-d4TMP-treated CEM cells was ~3.5 h, and the intracellular ratios of d4TTP/d4TMP in cells exposed to cycloSal-d4TMP gradually increased from 1 to 3.4 upon prolonged incubation. Radiolabeled cycloSal-d4TMP could be separated as its two R(p) and S(p) diastereomers on high-performance liquid chromatography. The R(p) diastereomer of cycloSal-d4TMP was 3- to 7-fold more efficient in releasing d4TMP and generating d4TTP than the S(p) cycloSal-d4TMP diastereomer. This correlated well with the 5-fold more pronounced antiviral activity of the R(p) diastereomer versus the S(p) diastereomer. d4TMP is a poor substrate for the cytosolic 5'(3')-deoxyribonucleotidase (V(max)/K(m) for d4TMP: 0.08 of V(max)/K(m) for dTMP) and is only slowly hydrolyzed to d4T. This contributes to the efficient conversion of the prodrug of d4TTP.

Original languageEnglish
Pages (from-to)928-935
Number of pages8
JournalMolecular Pharmacology
Volume58
Issue number5
Publication statusPublished - 2000

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Prodrugs
Antiviral Agents
Nucleotides
Cell Culture Techniques
2',3'-dideoxy-2',3'-didehydrothymidine monophosphate
Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate
Thymidine Kinase
Virus Replication
Half-Life
Hydrolysis
High Pressure Liquid Chromatography
HIV
Cell Line
5'(3')-nucleotidase

ASJC Scopus subject areas

  • Pharmacology

Cite this

Balzarini, J., Aquaro, S., Knispel, T., Rampazzo, C., Bianchi, V., Perno, C. F., ... Meier, C. (2000). Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate: Efficient intracellular delivery of d4TMP. Molecular Pharmacology, 58(5), 928-935.

Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate : Efficient intracellular delivery of d4TMP. / Balzarini, Jan; Aquaro, Stefano; Knispel, Tina; Rampazzo, Chiara; Bianchi, Vera; Perno, Carlo Federico; De Clercq, Erik; Meier, Chris.

In: Molecular Pharmacology, Vol. 58, No. 5, 2000, p. 928-935.

Research output: Contribution to journalArticle

Balzarini, J, Aquaro, S, Knispel, T, Rampazzo, C, Bianchi, V, Perno, CF, De Clercq, E & Meier, C 2000, 'Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate: Efficient intracellular delivery of d4TMP', Molecular Pharmacology, vol. 58, no. 5, pp. 928-935.
Balzarini, Jan ; Aquaro, Stefano ; Knispel, Tina ; Rampazzo, Chiara ; Bianchi, Vera ; Perno, Carlo Federico ; De Clercq, Erik ; Meier, Chris. / Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate : Efficient intracellular delivery of d4TMP. In: Molecular Pharmacology. 2000 ; Vol. 58, No. 5. pp. 928-935.
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abstract = "Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine-5'-monophosphate (cycloSal-d4TMP) is a potent and selective inhibitor of human immunodeficiency virus replication in cell culture and differs from other nucleotide prodrug approaches in that it is designed to selectively deliver the nucleotide 5'-monophosphate by a controlled, chemically induced hydrolysis. Its antiviral efficacy in cell culture is at least as good as, if not superior to, that of d4T. CycloSal-d4TMP was found to lead to the efficient intracellular release of d4TMP in a variety of cell lines, including both wild-type CEM and thymidine kinase-deficient CEM/TK- cells. Under similar experimental conditions, exposure of CEM/TK- cells to d4T failed to result in significant d4TTP levels. The intracellular conversion of cycloSal-d4TMP proved to be both time and dose dependent. The half-life of d4TTP generated intracellularly from d4T- or cycloSal-d4TMP-treated CEM cells was ~3.5 h, and the intracellular ratios of d4TTP/d4TMP in cells exposed to cycloSal-d4TMP gradually increased from 1 to 3.4 upon prolonged incubation. Radiolabeled cycloSal-d4TMP could be separated as its two R(p) and S(p) diastereomers on high-performance liquid chromatography. The R(p) diastereomer of cycloSal-d4TMP was 3- to 7-fold more efficient in releasing d4TMP and generating d4TTP than the S(p) cycloSal-d4TMP diastereomer. This correlated well with the 5-fold more pronounced antiviral activity of the R(p) diastereomer versus the S(p) diastereomer. d4TMP is a poor substrate for the cytosolic 5'(3')-deoxyribonucleotidase (V(max)/K(m) for d4TMP: 0.08 of V(max)/K(m) for dTMP) and is only slowly hydrolyzed to d4T. This contributes to the efficient conversion of the prodrug of d4TTP.",
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