title: In vitro pharmacokinetics of phosphorothioate antisense oligonucleotides.
abstract: ISIS 2105 (Afovirsen), a 20-mer phosphorothioate oligonucleotide that inhibits the production of a gene product essential to the growth of human papillomavirus, is in phase II clinical trials for the treatment of genital warts induced by human papillomavirus-6 and human papillomavirus-11. The uptake, subcellular distribution and metabolism of ISIS 2105 and three other similar length phosphorothioates have been studied in a variety of cell lines. Our experiments indicated that ISIS 2105 and other phosphorothioates are internalized and distributed in a time-, temperature-, concentration-, sequence- and cell line-dependent manner. Cell association was also influenced by the tissue culture medium. Several different analytical techniques revealed that phosphorothioates were more rapidly degraded in vitro than previously reported. These data suggest that phosphorothioate oligonucleotide uptake and stability observed in tissue culture can vary as a function of cellular assay conditions and analytical methods used. Comparison of these results with those obtained in vivo suggests that the pharmacokinetic behavior of this class of compounds cannot necessarily be predicted from in vitro studies.
authors.affiliation: Isis Pharmaceuticals, Inc., Carlsbad Research Center, California, USA.
authors.forename: R M
authors.initials: RM
authors.lastname: Crooke
authors.forename: M J
authors.initials: MJ
authors.lastname: Graham
authors.forename: M E
authors.initials: ME
authors.lastname: Cooke
authors.forename: S T
authors.initials: ST
authors.lastname: Crooke
mesh_terms: D000818:Animals; D000838:Anions; D000998:Antiviral Agents / Q000378:metabolism / Q000493:pharmacokinetics; D001345:Autoradiography; D001483:Base Sequence; D002250:Carbon Radioisotopes; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004355:Drug Stability; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000378:metabolism / Q000493:pharmacokinetics*; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000378:metabolism; D013873:Thionucleotides / Q000378:metabolism / Q000493:pharmacokinetics*; D014316:Tritium
chemical_list: D000838:Anions; D000998:Antiviral Agents; D002250:Carbon Radioisotopes; D003470:Culture Media; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D014316:Tritium; C084960:afovirsen sodium; D011116:Polynucleotide 5'-Hydroxyl-Kinase