Alexandra Dittmer1, Ina Woskobojnik1, Rebekka Adfeldt1, John C Drach2, Leroy B Townsend3, Sebastian Voigt4, Elke Bogner5. 1. Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany. 2. School of Dentistry, University of Michigan, Ann Arbor, USA; College of Pharmacy, University of Michigan, Ann Arbor, USA. 3. College of Pharmacy, University of Michigan, Ann Arbor, USA; Department of Chemistry, University of Michigan, Ann Arbor, USA. 4. Robert Koch-Institut, Berlin, Klinik für Pädiatrie mit Schwerpunkt Onkologie und Hämatologie, Charité -Universitätsmedizin Berlin, Berlin, Germany. 5. Institute of Medical Virology, Charité-Universitätsmedizin Berlin, Berlin, Germany. Electronic address: elke.bogner@charite.de.
Abstract
BACKGROUND: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
BACKGROUND: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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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: Vadim Makarov; Elke Bogner; Rebekka Adfeldt; Janna Schmitz; Barbara Kropff; Marco Thomas; Natalia Monakhova; Julia E Hölper; Barbara G Klupp; Thomas C Mettenleiter Journal: Antimicrob Agents Chemother Date: 2021-03-18 Impact factor: 5.191
Authors: Lana F Thaljeh; J Ainsley Rothschild; Misagh Naderi; Lyndon M Coghill; Jeremy M Brown; Michal Brylinski Journal: Biomolecules Date: 2019-10-12