Antiviral activities and phosphorylation of 5-halo-2'-deoxyuridines and N-methanocarbathymidine in cells infected with vaccinia virus.
BACKGROUND
The antipoxviral activities and phosphorylation of N-methanocarbathymidine ([N]-MCT) and four 5-halo-2'-deoxyuridines, namely 5-fluoro-(FdU), 5-chloro-(CldU), 5-bromo-(BrdU), and 5-iodo-(IdU) derivatives, were explored.


METHODS
Antiviral activities and nucleoside metabolism were determined in C127I mouse, LLC-MK2 monkey, and A549 human cells infected with thymidine-kinase-containing and -deficient (TK+ and TK-) vaccinia (WR strain) viruses.


RESULTS
The antiviral potencies of CldU, BrdU and IdU were increased 16-26-fold in LLC-MK2 cells infected with TK+ compared with TK- virus infections, but enhancement of activity was much less in the other cell lines. (N)-MCT was nearly equally active against TK+ and TK- viruses in the three cell lines. Antiviral activity of FdU was associated with cytotoxicity. Uninfected and infected cells metabolized compounds to mono-, di- and triphosphates. The thymidine, BrdU and IdU triphosphate levels were higher in C127I and LLC-MK2 cells infected with TK+ than with TK- virus. (N)-MCT monophosphate levels were much higher in TK+ virus-infected cells, but without corresponding increases in (N)-MCT triphosphate. Furthermore, TK+ virus infections did not appreciably alter (N)-MCT triphosphate levels in other mouse (L929), monkey (MA-104 and Vero) and human cell lines (A549). Antiviral potency of the compounds was greater in C127I than in LLC-MK2 cells, yet lower intracellular triphosphate levels were found in C127I cells.


CONCLUSIONS
We conclude that viral TK plays an important role in increasing the antiviral potencies of these compounds in some cell lines, but minimally in others. These findings may have implications in treating infected animals with compounds that are dependent upon poxvirus TK for their activation, because viral TK activity may vary greatly due to cell type.
Smee
Donald F
DF
Institute for Antiviral Research, Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, USA. dsmee@cc.usu.edu
Humphreys
Daniel E
DE
Hurst
Brett L
BL
Barnard
Dale L
DL
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D001973:Bromodeoxyuridine / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D005467:Floxuridine / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007065:Idoxuridine / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D010766:Phosphorylation; D013936:Thymidine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D013937:Thymidine Kinase / Q000378:metabolism; D014615:Vaccinia / Q000188:drug therapy*; D014616:Vaccinia virus / Q000187:drug effects*; D014709:Vero Cells; D010948:Viral Plaque Assay
D000998:Antiviral Agents; D005467:Floxuridine; C012244:5-chloro-2'-deoxyuridine; D013937:Thymidine Kinase; D001973:Bromodeoxyuridine; D007065:Idoxuridine; C469995:(north)-methanocarbathymidine; D013936:Thymidine; D003857:Deoxyuridine