Indolocarbazoles exhibit strong antiviral activity against human cytomegalovirus and are potent inhibitors of the pUL97 protein kinase.
We have analyzed a panel of protein kinase inhibitors (PKIs) and found that some indolocarbazoles (Gö6976, K252a, K252c) proved to be highly effective inhibitors of GCV-sensitive and -resistant human cytomegalovirus (HCMV) strains, but did not show any effect against herpes simplex virus. Antiviral activity was determined by focus reduction assays (IC(50) ranging from 0.009 to 0.4 microM). Other inhibitors of serine/threonine kinases (Gö6850, H-7, roscovitine) were found to be ineffective. Virus yield at 5 days after infection was reduced by three orders of magnitude with nanomolar concentrations of the indolocarbazoles. These compounds were fully effective when added up to 24 h post infection and showed reduced activity up to 72 h post infection. Cytotoxicity assays in proliferating and non-proliferating cells demonstrated that the effective antiviral concentration of these compounds was significantly lower than either antiproliferative (IC(50)/CC(50) ranging from 6.5 to 390) or cytotoxic (IC(50)/CC(50) ranging from 72. 5 to 1000) doses. The effects of PKIs on the virus-encoded protein kinase pUL97 were studied using recombinant vaccinia viruses. Indolocarbazoles strongly inhibited both pUL97 autophosphorylation (IC(50) ranging from 0.0012 to 0.013 microM) and pUL97-dependent ganciclovir phosphorylation (IC(50) ranging from 0.05 to 0.26 microM). Other inhibitors of serine/threonine kinases showed only weak (Gö6850) or no (H-7, roscovitine) effect on these pUL97 functions, while oxoflavone tyrosine kinase inhibitors had no effect at all.
Zimmermann
A
A
Abteilung Virologie, Institut für Mikrobiologie, Universität Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany.
Wilts
H
H
Lenhardt
M
M
Hahn
M
M
Mertens
T
T
D000998:Antiviral Agents / Q000494:pharmacology*; D002227:Carbazoles / Q000494:pharmacology*; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000821:virology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015774:Ganciclovir / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D026121:Indole Alkaloids; D007211:Indoles / Q000494:pharmacology; D008826:Microbial Sensitivity Tests / Q000379:methods; D009499:Neutral Red / Q000378:metabolism; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D018139:Simplexvirus / Q000187:drug effects; D014771:Virion / Q000187:drug effects / Q000201:enzymology; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D002227:Carbazoles; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D007211:Indoles; D047428:Protein Kinase Inhibitors; C081021:Go 6976; D009499:Neutral Red; C049985:staurosporine aglycone; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; D015774:Ganciclovir