title: Establishment of a cell-based assay for screening of compounds inhibiting very early events in the cytomegalovirus replication cycle and characterization of a compound identified using the assay.
abstract: To simplify the detection of infectious human cytomegalovirus (HCMV), we generated a cell line that produced luciferase in a dose-dependent manner upon HCMV infection. Using this cell line, we identified anti-HCMV compounds from a diverse library of 9,600 compounds. One of them, 1-(3,5-dichloro-4-pyridyl)piperidine-4-carboxamide (DPPC), was effective against HCMV (Towne strain) infection of human lung fibroblast cells at a 50% effective concentration of 2.5 microM. DPPC also inhibited the growth of clinical HCMV isolates and guinea pig and mouse cytomegaloviruses. Experiments using various time frames for treatment of the cells with DPPC demonstrated that DPPC was effective during the first 24 h after HCMV infection. DPPC treatment decreased not only viral DNA replication but also IE1 and IE2 expression at mRNA and protein levels in the HCMV-infected cells. However, DPPC did not inhibit the attachment of HCMV particles to the cell surface. DPPC is a unique compound that targets the very early phase of cytomegalovirus infection, probably by disrupting a pathway that is important after viral entry but before immediate-early gene expression.
authors.affiliation: Department of Virology I, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Fukui
authors.forename: Keiko
authors.initials: K
authors.lastname: Shindoh
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Shin
authors.initials: S
authors.lastname: Koyano
authors.forename: Isao
authors.initials: I
authors.lastname: Kosugi
authors.forename: Toyofumi
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Ichiro
authors.initials: I
authors.lastname: Kurane
authors.forename: Naoki
authors.initials: N
authors.lastname: Inoue
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D004261:DNA Replication / Q000187:drug effects; D017781:Genes, Immediate-Early / Q000187:drug effects; D006168:Guinea Pigs; D006801:Humans; D051379:Mice; D008826:Microbial Sensitivity Tests / Q000379:methods*; D018146:Muromegalovirus / Q000187:drug effects / Q000502:physiology; D010880:Piperidines / Q000737:chemistry / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D019211:Roseolovirus / Q000187:drug effects / Q000502:physiology; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D010880:Piperidines; D012333:RNA, Messenger; D012367:RNA, Viral