title: Tetrahalogenated benzimidazole D-ribonucleosides are active against rat cytomegalovirus.
abstract: Benzimidazole D-ribonucleosides are potent and selective inhibitors of CMV infection that have been shown to target the viral terminase, the enzyme complex responsible for viral DNA cleavage into single unit-length genomes and subsequent DNA packaging into procapsids. Here, we evaluated the viral inhibition by benzimidazole D-ribonucleosides against rat cytomegalovirus (RCMV).
  
  
  
  Antiviral activity of compounds Cl4RB and BTCRB against RCMV was quantified by measurement of plaque formation. Yield assays and electron microscopy of thin sections was performed using RCMV-infected cells in the presence or absence of the compounds. The effects of Cl4RB and BTCRB on cleavage of concatemers was analyzed by pulsed-field gel electrophoresis. To characterize the behaviour of the antiviral compounds in a more physiological environment, a 3D cell culture model was employed where cells are embedded in an extracellular matrix using rat-tail collagen I.
  
  
  
  Both compounds had an inhibitory effect against RCMV-E. Electron microscopy revealed that only few virions were formed in RCMV-E infected cells in the presence of the compounds. Pulsed-field gel electrophoresis showed that DNA concatemers failed to be processed in the presence of the compounds. Yield Assays showed a comparable viral growth in the 3D vs. 2D cell culture as well as inhibition in the presence of Cl4RB or BTCRB for RCMV-E/GFP.
  
  
  
  These results demonstrate that the tetrahalogenated benzimidazole D-ribonucleosides are effective against RCMV-E by preventing cleavage of concatemeric DNA and nuclear egress of mature capsids.
authors.affiliation: Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Dittmer
authors.affiliation: Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Ina
authors.initials: I
authors.lastname: Woskobojnik
authors.affiliation: Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Rebekka
authors.initials: R
authors.lastname: Adfeldt
authors.affiliation: School of Dentistry, University of Michigan, Ann Arbor, USA; College of Pharmacy, University of Michigan, Ann Arbor, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: Drach
authors.affiliation: College of Pharmacy, University of Michigan, Ann Arbor, USA; Department of Chemistry, University of Michigan, Ann Arbor, USA.
authors.forename: Leroy B
authors.initials: LB
authors.lastname: Townsend
authors.affiliation: Robert Koch-Institut, Berlin, Klinik für Pädiatrie mit Schwerpunkt Onkologie und Hämatologie, Charité -Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Voigt
authors.affiliation: Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany. Electronic address: elke.bogner@charite.de.
authors.forename: Elke
authors.initials: E
authors.lastname: Bogner
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D001562:Benzimidazoles / Q000737:chemistry / Q000494:pharmacology*; D018929:Cell Culture Techniques; D003094:Collagen / Q000737:chemistry; D042003:DNA Packaging / Q000187:drug effects; D004706:Endodeoxyribonucleases / Q000187:drug effects; D054879:Halogenation; D006566:Herpesviridae Infections / Q000188:drug therapy* / Q000821:virology; D008854:Microscopy, Electron; D008954:Models, Biological; D018146:Muromegalovirus / Q000187:drug effects* / Q000648:ultrastructure; D009705:Nucleosides / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D012263:Ribonucleosides / Q000737:chemistry / Q000494:pharmacology*; D054457:Tissue Scaffolds; D010948:Viral Plaque Assay
chemical_list: C000625402:2,4,5,6-tetrachloro-1-(2,3,5-tri-O-acetylribofuranosyl)benzimidazole; C000625401:2-bromo-4,5,6-trichloro-1-(2,3,5-tri-O-acetylribofuranosyl)benzimidazole; D000998:Antiviral Agents; D001562:Benzimidazoles; D009705:Nucleosides; D012263:Ribonucleosides; D003094:Collagen; D004706:Endodeoxyribonucleases; C030894:terminase
keywords: Antiviral activity; Cleavage of concatemers; DNA packaging; Rat cytomegalovirus; Tetrahalogenated benzimidazoles