Enhancing antisense efficacy with multimers and multi-targeting oligonucleotides (MTOs) using cleavable linkers

• Published in: Nucleic acids research Vol. 43; no. 19; pp. 9123 - 9132
• Main Authors: Subramanian, Romesh R, Wysk, Mark A, Ogilvie, Kathleen M, Bhat, Abhijit, Kuang, Bing, Rockel, Thomas D, Weber, Markus, Uhlmann, Eugen, Krieg, Arthur M
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 30-10-2015
• Subjects: Animals; Apolipoprotein C-III - genetics; Apolipoprotein C-III - metabolism; Apolipoproteins B - genetics; Apolipoproteins B - metabolism; Chemical Biology and Nucleic Acid Chemistry; Dimerization; Female; Humans; Liver - metabolism; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; MicroRNAs - antagonists & inhibitors; Oligonucleotides, Antisense - chemistry; Oligonucleotides, Antisense - pharmacokinetics; Oligonucleotides, Antisense - pharmacology; Tissue Distribution
• DOI: 10.1093/nar/gkv992
• EISSN: 1362-4962
• ISSN: 0305-1048

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.