Mode of antiviral action of penciclovir in MRC-5 cells infected with herpes simplex virus type 1 (HSV-1), HSV-2, and varicella-zoster virus.
The metabolism and mode of action of penciclovir [9-(4-hydroxy-3-hydroxymethylbut-1-yl)guanine; BRL 39123] were studied and compared with those of acyclovir. In uninfected MRC-5 cells, low concentrations of the triphosphates of penciclovir and acyclovir were occasionally just detectable, the limit of detection being about 1 pmol/10(6) cells. In contrast, in cells infected with either herpes simplex virus type 2 (HSV-2) or varicella-zoster virus (VZV), penciclovir was phosphorylated quickly to give high concentrations of the triphosphate ester. Following the removal of penciclovir from the culture medium, penciclovir-triphosphate remained trapped within the cells for a long time (half-lives, 20 and 7 h in HSV-2- and VZV-infected cells, respectively). In HSV-2-infected cells, acyclovir was phosphorylated to a lesser extent and the half-life of the triphosphate ester was only 1 h. We were unable to detect any phosphates of acyclovir in VZV-infected cells. (S)-Penciclovir-triphosphate inhibited HSV-1 and HSV-2 DNA polymerase competitively with dGTP, the Ki values being 8.5 and 5.8 microM, respectively, whereas for acyclovir-triphosphate, the Ki value was 0.07 microM for the two enzymes. Both compounds had relatively low levels of activity against the cellular DNA polymerase alpha, with Ki values of 175 and 3.8 microM, respectively. (S)-Penciclovir-triphosphate did inhibit DNA synthesis by HSV-2 DNA polymerase with a defined template-primer, although it was not an obligate chain terminator like acyclovir-triphosphate. These results provide a biochemical rationale for the highly selective and effective inhibition of HSV-2 and VZV DNA synthesis by penciclovir and for the greater activity of penciclovir than that of acyclovir when HSV-2-infected cells were treated for a short time.
Earnshaw
D L
DL
SmithKline Beecham Pharmaceuticals, Epsom, Surrey, England.
Bacon
T H
TH
Darlison
S J
SJ
Edmonds
K
K
Perkins
R M
RM
Vere Hodge
R A
RA
D000212:Acyclovir / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification; D004951:Esterification; D006147:Guanine; D006561:Herpes Simplex / Q000378:metabolism / Q000382:microbiology; D006562:Herpes Zoster / Q000378:metabolism / Q000382:microbiology; D014645:Herpesvirus 3, Human / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D019384:Nucleic Acid Synthesis Inhibitors; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D018139:Simplexvirus / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D013997:Time Factors
D004279:DNA, Viral; D019384:Nucleic Acid Synthesis Inhibitors; D010710:Phosphates; C053539:penciclovir; D006147:Guanine; D004259:DNA-Directed DNA Polymerase; D000212:Acyclovir