title: Enhancing antisense efficacy with multimers and multi-targeting oligonucleotides (MTOs) using cleavable linkers.
abstract: The in vivo potency of antisense oligonucleotides (ASO) has been significantly increased by reducing their length to 8-15 nucleotides and by the incorporation of high affinity RNA binders such as 2', 4'-bridged nucleic acids (also known as locked nucleic acid or LNA, and 2',4'-constrained ethyl [cET]). We now report the development of a novel ASO design in which such short ASO monomers to one or more targets are co-synthesized as homo- or heterodimers or multimers via phosphodiester linkers that are stable in plasma, but cleaved inside cells, releasing the active ASO monomers. Compared to current ASOs, these multimers and multi-targeting oligonucleotides (MTOs) provide increased plasma protein binding and biodistribution to liver, and increased in vivo efficacy against single or multiple targets with a single construct. In vivo, MTOs synthesized in both RNase H-activating and steric-blocking oligonucleotide designs provide ≈4-5-fold increased potency and ≈2-fold increased efficacy, suggesting broad therapeutic applications.
authors.affiliation: RaNA Therapeutics LLC, 790 Memorial Dr., Suite 203, Cambridge, MA 02139, USA.
authors.forename: Romesh R
authors.initials: RR
authors.lastname: Subramanian
authors.affiliation: RaNA Therapeutics LLC, 790 Memorial Dr., Suite 203, Cambridge, MA 02139, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Wysk
authors.affiliation: Pfizer Worldwide Research and Development, 9381 Judicial Dr., Suite 200, San Diego, CA 92121, USA.
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Ogilvie
authors.affiliation: Pfizer Worldwide Research and Development, 9381 Judicial Dr., Suite 200, San Diego, CA 92121, USA.
authors.forename: Abhijit
authors.initials: A
authors.lastname: Bhat
authors.affiliation: Pfizer Worldwide Research and Development, 9381 Judicial Dr., Suite 200, San Diego, CA 92121, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Kuang
authors.affiliation: Coley Pharmaceutical GmbH, Merowingerplatz 1a, 40225 Dusseldorf, Germany.
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Rockel
authors.affiliation: Coley Pharmaceutical GmbH, Merowingerplatz 1a, 40225 Dusseldorf, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Weber
authors.affiliation: Coley Pharmaceutical GmbH, Merowingerplatz 1a, 40225 Dusseldorf, Germany.
authors.forename: Eugen
authors.initials: E
authors.lastname: Uhlmann
authors.affiliation: RaNA Therapeutics LLC, 790 Memorial Dr., Suite 203, Cambridge, MA 02139, USA cpgrocks@gmail.com.
authors.forename: Arthur M
authors.initials: AM
authors.lastname: Krieg
mesh_terms: D000818:Animals; D053305:Apolipoprotein C-III / Q000235:genetics / Q000378:metabolism; D001055:Apolipoproteins B / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D005260:Female; D006801:Humans; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000037:antagonists & inhibitors; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D014018:Tissue Distribution
chemical_list: D053305:Apolipoprotein C-III; D001055:Apolipoproteins B; D035683:MicroRNAs; C544354:Mirn122 microRNA, mouse; D016376:Oligonucleotides, Antisense