title: Variations in mRNA content have no effect on the potency of antisense oligonucleotides.
abstract: A fundamental question with regard to antisense pharmacology is the extent to which RNA content or transcription rate or both affect the potency of antisense drugs. We have addressed this by controlling RNA content and transcription rate using either an exogenous gene expressed after transfection or an endogenous gene induced with a cytokine. We have demonstrated that in both A549 and HeLa cells, varying RNA copy numbers from <1 to >100 copies per cell has no effect on the potency of RNase H-active antisense drugs transfected into cells, nor did variation in transcription rate have an effect on potency. We demonstrate that this is because the number of oligonucleotide molecules per cell is vastly in excess of the RNA copy number. These data further suggest that a significant fraction of cell-associated antisense drug molecules may be unavailable to interact with the target RNA, an observation that is not surprising, as phosphorothioate oligonucleotides interact with many cellular proteins. We suggest that these data may extrapolate to in vivo results.
authors.affiliation: Isis Pharmaceuticals, Inc., Carlsbad, CA 92008, USA.
authors.forename: L
authors.initials: L
authors.lastname: Miraglia
authors.forename: A T
authors.initials: AT
authors.lastname: Watt
authors.forename: M J
authors.initials: MJ
authors.lastname: Graham
authors.forename: S T
authors.initials: ST
authors.lastname: Crooke
mesh_terms: D003907:Dexamethasone / Q000494:pharmacology; D004347:Drug Interactions; D015870:Gene Expression / Q000187:drug effects*; D018390:Gene Targeting; D017930:Genes, Reporter; D011905:Genes, ras / Q000187:drug effects / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000037:antagonists & inhibitors / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D054735:Phosphorothioate Oligonucleotides; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C122000:ISIS 2503; D016376:Oligonucleotides, Antisense; D054735:Phosphorothioate Oligonucleotides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D003907:Dexamethasone; D008156:Luciferases