Mesyl Phosphoramidate Oligonucleotides as Potential Splice-Switching Agents: Impact of Backbone Structure on Activity and Intracellular Localization.

Hammond SM., Sergeeva OV., Melnikov PA., Goli L., Stoodley J., Zatsepin TS., Stetsenko DA., Wood MJA.

A series of 2'-deoxy and novel 2'-O-methyl and 2'-O-(2-methoxyethyl) (2'-MOE) oligonucleotides with internucleotide methanesulfonyl (mesyl, μ) or 1-butanesulfonyl (busyl, β) phosphoramidate groups has been synthesized for evaluation as potential splice-switching oligonucleotides. Evaluation of their splice-switching activity in spinal muscular atrophy patient-derived fibroblasts revealed no significant difference in splice-switching efficacy between 2'-MOE mesyl oligonucleotide and the corresponding phosphorothioate (nusinersen). Yet, a survival study with model neonatal mice has shown the antisense 2'-MOE mesyl oligonucleotide to be inferior to nusinersen at the highest dose of 40 mg/kg. A reason for their lower activity in vivo as ascertained by cellular uptake study by fluorescent confocal microscopy in HEK293 cell line could possibly be ascribed to compromised endosomal release and/or nuclear uptake of the 2'-OMe or 2'-MOE μ- and β-oligonucleotides compared to their phosphorothioate analog.

DOI

10.1089/nat.2020.0860

Type

Journal article

Publication Date

2021-06-01T00:00:00+00:00

Volume

31

Pages

190 - 200

Total pages

10

Keywords

2′-methoxyethyl, intracellular distribution, nusinersen, spinal muscular atrophy, splice switching, Amides, Animals, HEK293 Cells, Humans, Mice, Muscular Atrophy, Spinal, Oligonucleotides, Oligonucleotides, Antisense, Phosphoric Acids

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