Characterization of the Activation Pathway of Phosphoramidate Triester Prodrugs of Stavudine and Zidovudine

The phosphoramidate triester prodrugs of anti-human HIV 2′,3′-dideoxynucleoside analogs (ddN) represent a convenient approach to bypass the first phosphorylation to ddN 5′-monophosphate (ddNMP), resulting in an improved formation of ddN 5′-triphosphate and, hence, higher antiviral efficacy....

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Molecular pharmacology 1999-10, Vol.56 (4), p.693-704
Hauptverfasser: Saboulard, D, Naesens, L, Cahard, D, Salgado, A, Pathirana, R, Velazquez, S, McGuigan, C, De Clercq, E, Balzarini, J
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung:The phosphoramidate triester prodrugs of anti-human HIV 2′,3′-dideoxynucleoside analogs (ddN) represent a convenient approach to bypass the first phosphorylation to ddN 5′-monophosphate (ddNMP), resulting in an improved formation of ddN 5′-triphosphate and, hence, higher antiviral efficacy. Although phosphoramidate derivatization markedly increases the anti-HIV activity of 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) in both wild-type and thymidine kinase-deficient CEM cells, the concept is far less successful for the 3′-azido-2′,3′-dideoxythymidine (AZT) triesters. We now investigated the metabolism of triester prodrugs of d4T and AZT using pure enzymes or different biological media. The efficiency of the first activation step, mediated by carboxylesterases, consists of the formation of the amino acyl ddNMP metabolite. The efficiency of this step was shown to be dependent on the amino acid, alkyl ester, and ddN moiety. Triesters that showed no conversion to the amino acyl ddNMP accumulated as the phenyl-containing intermediate and had poor, if any, anti-HIV activity. In contrast to the relative stability of the triesters in human serum, carboxylesterase-mediated cleavage of the prodrugs was found to be remarkably high in mouse serum. The subsequent conversion of the amino acyl ddNMP metabolite to ddNMP or ddN was highest in rat liver cytosolic enzyme preparations. Although l -alaninyl-d4TMP was efficiently converted to d4TMP, the main metabolite formed from l -alaninyl-AZTMP was the free nucleoside (AZT), thus explaining why d4T prodrugs, but not AZT prodrugs, retain anti-HIV activity in HIV-infected thymidine kinase-deficient cell cultures. The rat liver phosphoramidase responsible for the formation of ddNMP was shown to be distinct from creatine kinase, alkaline phosphatase, and phosphodiesterase.
ISSN:0026-895X
1521-0111