| Autor: |
Zhang L; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Liang XH; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., De Hoyos CL; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Migawa M; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Nichols JG; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Freestone G; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Tian J; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Seth PP; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA., Crooke ST; Department of Core Antisense Research, Ionis Pharmaceuticals, Inc., Carlsbad, California, USA. |
| Abstrakt: |
Antisense oligonucleotides (ASOs) that mediate RNA target degradation by RNase H1 are used as drugs to treat various diseases. Previously we found that introduction of a single 2'- O -methyl (2'-OMe) modification in position 2 of the central deoxynucleotide region of a gapmer phosphorothioate (PS) ASO, in which several residues at the termini are 2'-methoxyethyl, 2' constrained ethyl, or locked nucleic acid, dramatically reduced cytotoxicity with only modest effects on potency. More recently, we demonstrated that replacement of the PS linkage at position 2 or 3 in the gap with a mesyl-phosphoramidate (MsPA) linkage also significantly reduced toxicity without meaningful loss of potency and increased the elimination half-life of the ASOs. In this study, we evaluated the effects of the combination of MsPA linkages and 2'-OMe nucleotides on PS ASO performance. We found that two MsPA modifications at the 5' end of the gap or in the 3'-wing of a Gap 2'-OMe PS ASO substantially increased the activity of ASOs with OMe at position 2 of the gap without altering the safety profile. Such effects were observed with multiple sequences in cells and animals. Thus, the MsPA modification improves the RNase H1 cleavage rate of PS ASOs with a 2'-OMe in the gap, significantly reduces binding of proteins involved in cytotoxicity, and prolongs elimination half-lives. |
