Stereochemical considerations in the enzymatic phosphorylation and antiviral activity of acyclonucleosides. I. Phosphorylation of 2'-nor-2'-deoxyguanosine.
The antiviral compound 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (2'-nor-2'-deoxyguanosine, 2'-NDG) is phosphorylated by the HSV-1-induced thymidine kinase to the monophosphate (2'-NDG-MP) and this is further phosphorylated by cellular kinases to the triphosphate (2'-NDG-TP) which is a potent inhibitor of DNA polymerases. Since phosphorylation of 2'-NDG creates a chiral center in the molecule, it was of interest to examine whether both monophosphate enantiomers were produced by the viral thymidine kinase, whether they both could be further phosphorylated by cellular kinases and, if so, whether the respective triphosphates were equally inhibitory to the DNA polymerases. The time course of the phosphorylation by GMP kinase of a chemically synthesized, racemic 2'-NDG-MP was compared to that of a 2'-NDG-MP preparation obtained by enzymatic phosphorylation of 2'-NDG with HSV-1 thymidine kinase. The results indicated that the two enantiomeric monophosphates were phosphorylated by GMP kinase with different rates and that phosphorylation of 2'-NDG by HSV-1 thymidine kinase gave only one of the isomers, whose structure was determined to be S. Both enantiomeric diphosphates were further phosphorylated to the respective triphosphates and it was shown that, in contrast to the triphosphate obtained from the 2'-NDG-MP prepared by viral thymidine kinase which was a potent inhibitor of HSV-1 DNA polymerase, the triphosphate obtained from the slow-reacting R isomer had little or no inhibitory activity against this enzyme.
D000212:Acyclovir / Q000031:analogs & derivatives* / Q000378:metabolism; D015774:Ganciclovir; D051528:Guanylate Kinases; D007700:Kinetics; D009703:Nucleoside-Phosphate Kinase / Q000378:metabolism; D010504:Periodic Acid / Q000378:metabolism; D018139:Simplexvirus / Q000378:metabolism; D013237:Stereoisomerism; D013937:Thymidine Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D010504:Periodic Acid; D013937:Thymidine Kinase; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases; D015774:Ganciclovir; D000212:Acyclovir