A novel nonnucleoside inhibitor specifically targets cytomegalovirus DNA maturation via the UL89 and UL56 gene products.
3-Hydroxy-2,2-dimethyl-N-[4([[5-(dimethylamino)-1-naphthyl]sulfonyl]amino)-phenyl]propanamide (BAY 38-4766) is a novel selective nonnucleoside inhibitor of cytomegalovirus (CMV) replication with an excellent safety profile. This compound and structural analogues inhibit neither viral DNA synthesis nor viral transcription and translation. Accumulation of dense bodies and noninfectious enveloped particles coincides with inhibition of both concatemer processing and functional cleavage at intergenomic transitions, pointing to interference with viral DNA maturation and packaging of monomeric genome lengths. Resistant virus populations, including a murine CMV (MCMV) isolate with 566-fold-decreased drug sensitivity, were selected in vitro. Sequencing of the six open reading frames (ORFs) known to be essentially involved in viral DNA cleavage and packaging identified mutations in ORFs UL56, UL89, and UL104. Construction of MCMV recombinants expressing different combinations of murine homologues of mutant UL56, UL89, and UL104 and analysis of drug susceptibilities clearly demonstrated that mutant ORFs UL89 exon II (M360I) and M56 (P202A I208N) individually confer resistance to BAY 38-4766. A combination of both mutant proteins exhibited a strong synergistic effect on resistance, reconstituting the high-resistance phenotype of the in vitro mutant. These findings are consistent with genetic mapping of resistance to TCRB (2,5,6-trichloro-1-beta-D-ribofuranosyl benzimidazole) (P. M. Krosky et al., J. Virol. 72:4721-4728, 1998) and provide further indirect evidence that proteins encoded by UL89 and UL56 function as two subunits of the CMV terminase. While these studies also suggest that the molecular mechanism of BAY 38-4766 is distinct from that of benzimidazole ribonucleosides, they also offer an explanation for the excellent specificity and tolerability of BAY 38-4766, since mammalian DNA does not undergo comparable maturation steps.
Buerger
I
I
Antiinfective Research, Virology, Business Group Pharma, Bayer AG, D-42096 Wuppertal, Germany.
Reefschlaeger
J
J
Bender
W
W
Eckenberg
P
P
Popp
A
A
Weber
O
O
Graeper
S
S
Klenk
H D
HD
Ruebsamen-Waigmann
H
H
Hallenberger
S
S
D000818:Animals; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000188:drug therapy; D004279:DNA, Viral / Q000187:drug effects / Q000235:genetics; D006801:Humans; D051379:Mice; D009282:Naphthalenesulfonates / Q000494:pharmacology* / Q000627:therapeutic use; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D019065:Virus Assembly / Q000187:drug effects*
C434758:3-hydroxy-2,2-dimethyl-N-(4(((5-(dimethylamino)-1-naphthyl)sulfonyl)amino)phenyl)propanamide; D004279:DNA, Viral; D009282:Naphthalenesulfonates; C090238:UL56 protein, cytomegalovirus; C436448:UL89 protein, Cytomegalovirus; D014764:Viral Proteins; D015678:Viral Structural Proteins