A novel drug screening assay for papillomavirus specific antiviral activity.
Discovery and development of human papillomavirus (HPV) specific antiviral agents have been hampered by the lack of an in vitro assay permissive to HPV replication. An experimental assay system for monitoring HPV-11 DNA replication has been optimized for use as a papillomavirus antiviral drug screening tool. Cloned HPV DNA was introduced into SCC-4 cells by electroporation and viral DNA replication monitored by Southern blot. Kinetic studies demonstrated an increased HPV genome copy number with time. Viral DNA replicated as episomal, unit length genome and remained episomal after multiple passages. These data suggested the basis for an in vitro replication assay for evaluating the antiviral activity of potential chemotherapeutic agents directly on HPV. This model was used to investigate antiviral activities of current anti-HPV therapies such as 5-fluorouracil (5-FU) and alpha-interferon (alpha-IFN) and potential therapies such as sodium butyrate, 5-bromo-20-deoxyuridine (BrdU) and antisense oligonucleotides. HPV- 11 replication is significantly inhibited by BrdU and sodium butyrate; however 5-FU and alpha-IFN did not give consistent dose response results. Finally, ISIS 2105, a 20-mer phosphorothioate antisense oligonucleotide, which targets HPV-11 E2 gene product, showed potent antiviral activity in this assay with an IC50 of approximately 70 nM.
Clark
P R
PR
Department of Infectious Diseases, Isis Pharmaceuticals Inc., Carlsbad, CA 92008, USA.
Roberts
M L
ML
Cowsert
L M
LM
D000998:Antiviral Agents / Q000494:pharmacology*; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D007700:Kinetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D027383:Papillomaviridae / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D010957:Plasmids; D013873:Thionucleotides / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D002087:Butyrates; D004279:DNA, Viral; D016898:Interferon-alpha; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D020148:Butyric Acid; C084960:afovirsen sodium; D001973:Bromodeoxyuridine; D005472:Fluorouracil