title: Preclinical PK and PD studies on 2'-O-methyl-phosphorothioate RNA antisense oligonucleotides in the mdx mouse model.
abstract: Antisense oligonucleotides (AONs) are being developed as RNA therapeutic molecules for Duchenne muscular dystrophy. For oligonucleotides with the 2'-O-methyl-phosphorothioate (2OMePS) RNA chemistry, proof of concept has been obtained in patient-specific muscle cell cultures, the mouse and dog disease models, and recently by local administration in Duchenne patients. To further explore the pharmacokinetic (PK)/pharmacodynamic (PD) properties of this chemical class of oligonucleotides, we performed a series of preclinical studies in mice. The results demonstrate that the levels of oligonucleotides in dystrophin-deficient muscle fibers are much higher than in healthy fibers, leading to higher exon-skipping levels. Oligonucleotide levels and half-life differed for specific muscle groups, with heart muscle showing the lowest levels but longest half-life (approximately 46 days). Intravenous (i.v.), subcutaneous (s.c.), and intraperitoneal (i.p.) delivery methods were directly compared. For each method, exon-skipping and novel dystrophin expression were observed in all muscles, including arrector pili smooth muscle in skin biopsies. After i.v. administration, the oligonucleotide peak levels in plasma, liver, and kidney were higher than after s.c. or i.p. injections. However, as the bioavailability was similar, and the levels of oligonucleotide, exon-skipping, and dystrophin steadily accumulated overtime after s.c. administration, we selected this patient-convenient delivery method for future clinical study protocols.
authors.affiliation: Center for Human and Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Hans
authors.initials: H
authors.lastname: Heemskerk
authors.forename: Christa
authors.initials: C
authors.lastname: de Winter
authors.forename: Petra
authors.initials: P
authors.lastname: van Kuik
authors.forename: Niki
authors.initials: N
authors.lastname: Heuvelmans
authors.forename: Patrizia
authors.initials: P
authors.lastname: Sabatelli
authors.forename: Paola
authors.initials: P
authors.lastname: Rimessi
authors.forename: Paola
authors.initials: P
authors.lastname: Braghetta
authors.forename: Gert-Jan B
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: Sjef
authors.initials: S
authors.lastname: de Kimpe
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ferlini
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
authors.forename: Judith C T
authors.initials: JC
authors.lastname: van Deutekom
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004195:Disease Models, Animal; D016189:Dystrophin / Q000008:administration & dosage; D005455:Fluorescent Antibody Technique; D051379:Mice; D018101:Mice, Inbred mdx; D054735:Phosphorothioate Oligonucleotides / Q000493:pharmacokinetics / Q000494:pharmacology*; D016372:RNA, Antisense / Q000493:pharmacokinetics / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016189:Dystrophin; D054735:Phosphorothioate Oligonucleotides; D016372:RNA, Antisense
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