Substituted benzimidazoles with nanomolar activity against respiratory syncytial virus.
A cell-based assay was used to discover compounds inhibiting respiratory syncytial virus (RSV)-induced fusion in HeLa/M cells. A lead compound was identified and subsequent synthesis of >300 analogues led to the identification of JNJ 2408068 (R170591), a low molecular weight (MW 395) benzimidazole derivative with an EC(50) (0.16 nM) against some lab strains almost 100,000 times better than that of ribavirin (15 microM). Antiviral activity was confirmed for subgroup A and B clinical isolates of human RSV and for a bovine RSV isolate. The compound did not inhibit the growth of representative viruses from other Paramyxovirus genera, i.e. HPIV2 and Mumps Virus (genus Rubulavirus), HPIV3 (genus Respirovirus), Measles virus (genus Morbillivirus) and hMPV. Efficacy in cytopathic effect inhibition assays correlated well with efficacy in virus yield reduction assays. A concentration of 10nM reduced RSV production 1000-fold in multi-cycle experiments, irrespective of the multiplicity of infection. Time of addition studies pointed to a dual mode of action: inhibition of virus-cell fusion early in the infection cycle and inhibition of cell-cell fusion at the end of the replication cycle. Two resistant mutants were raised and shown to have single point mutations in the F-gene (S398L and D486N). JNJ 2408068 was also shown to inhibit the release of proinflammatory cytokines IL-6, IL-8 and Rantes from RSV-infected A549 cells.
Andries
Koen
K
Johnson and Johnson Pharmaceutical Research and Development, Beerse, Belgium.
Moeremans
Marc
M
Gevers
Tom
T
Willebrords
Rudy
R
Sommen
Cois
C
Lacrampe
Jean
J
Janssens
Frans
F
Wyde
Philip R
PR
D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D001562:Benzimidazoles / Q000737:chemistry* / Q000494:pharmacology*; D002459:Cell Fusion; D016207:Cytokines / Q000378:metabolism; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D004252:DNA Mutational Analysis; D024882:Drug Resistance, Viral / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D029121:Metapneumovirus / Q000187:drug effects / Q000254:growth & development; D008970:Molecular Weight; D018103:Morbillivirus / Q000187:drug effects / Q000254:growth & development; D017354:Point Mutation; D012136:Respiratory Syncytial Viruses / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification / Q000472:pathogenicity; D018108:Respirovirus / Q000187:drug effects / Q000254:growth & development; D019213:Rubulavirus / Q000187:drug effects / Q000254:growth & development; D014760:Viral Fusion Proteins / Q000235:genetics; D010948:Viral Plaque Assay
D000998:Antiviral Agents; D001562:Benzimidazoles; D016207:Cytokines; D014760:Viral Fusion Proteins