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Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages
Bicyclic oxazaphospholidine monomers were used to prepare a series of phosphorothioate (PS)-modified gapmer antisense oligonucleotides (ASOs) with control of the chirality of each of the PS linkages within the 10-base gap. The stereoselectivity was determined to be 98% for each coupling. The objecti...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4267618/ https://www.ncbi.nlm.nih.gov/pubmed/25398895 http://dx.doi.org/10.1093/nar/gku1115 |
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author | Wan, W. Brad Migawa, Michael T. Vasquez, Guillermo Murray, Heather M. Nichols, Josh G. Gaus, Hans Berdeja, Andres Lee, Sam Hart, Christopher E. Lima, Walt F. Swayze, Eric E. Seth, Punit P. |
author_facet | Wan, W. Brad Migawa, Michael T. Vasquez, Guillermo Murray, Heather M. Nichols, Josh G. Gaus, Hans Berdeja, Andres Lee, Sam Hart, Christopher E. Lima, Walt F. Swayze, Eric E. Seth, Punit P. |
author_sort | Wan, W. Brad |
collection | PubMed |
description | Bicyclic oxazaphospholidine monomers were used to prepare a series of phosphorothioate (PS)-modified gapmer antisense oligonucleotides (ASOs) with control of the chirality of each of the PS linkages within the 10-base gap. The stereoselectivity was determined to be 98% for each coupling. The objective of this work was to study how PS chirality influences biophysical and biological properties of the ASO including binding affinity (T(m)), nuclease stability, activity in vitro and in vivo, RNase H activation and cleavage patterns (both human and E. coli) in a gapmer context. Compounds that had nine or more Sp-linkages in the gap were found to be poorly active in vitro, while compounds with uniform Rp-gaps exhibited activity very similar to that of the stereo-random parent ASOs. Conversely, when tested in vivo, the full Rp-gap compound was found to be quickly metabolized resulting in low activity. A total of 31 ASOs were prepared with control of the PS chirally of each linkage within the gap in an attempt to identify favorable Rp/Sp positions. We conclude that a mix of Rp and Sp is required to achieve a balance between good activity and nuclease stability. |
format | Online Article Text |
id | pubmed-4267618 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-42676182014-12-23 Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages Wan, W. Brad Migawa, Michael T. Vasquez, Guillermo Murray, Heather M. Nichols, Josh G. Gaus, Hans Berdeja, Andres Lee, Sam Hart, Christopher E. Lima, Walt F. Swayze, Eric E. Seth, Punit P. Nucleic Acids Res Chemical Biology and Nucleic Acid Chemistry Bicyclic oxazaphospholidine monomers were used to prepare a series of phosphorothioate (PS)-modified gapmer antisense oligonucleotides (ASOs) with control of the chirality of each of the PS linkages within the 10-base gap. The stereoselectivity was determined to be 98% for each coupling. The objective of this work was to study how PS chirality influences biophysical and biological properties of the ASO including binding affinity (T(m)), nuclease stability, activity in vitro and in vivo, RNase H activation and cleavage patterns (both human and E. coli) in a gapmer context. Compounds that had nine or more Sp-linkages in the gap were found to be poorly active in vitro, while compounds with uniform Rp-gaps exhibited activity very similar to that of the stereo-random parent ASOs. Conversely, when tested in vivo, the full Rp-gap compound was found to be quickly metabolized resulting in low activity. A total of 31 ASOs were prepared with control of the PS chirally of each linkage within the gap in an attempt to identify favorable Rp/Sp positions. We conclude that a mix of Rp and Sp is required to achieve a balance between good activity and nuclease stability. Oxford University Press 2014-12-16 2014-11-14 /pmc/articles/PMC4267618/ /pubmed/25398895 http://dx.doi.org/10.1093/nar/gku1115 Text en © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Chemical Biology and Nucleic Acid Chemistry Wan, W. Brad Migawa, Michael T. Vasquez, Guillermo Murray, Heather M. Nichols, Josh G. Gaus, Hans Berdeja, Andres Lee, Sam Hart, Christopher E. Lima, Walt F. Swayze, Eric E. Seth, Punit P. Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title | Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title_full | Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title_fullStr | Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title_full_unstemmed | Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title_short | Synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
title_sort | synthesis, biophysical properties and biological activity of second generation antisense oligonucleotides containing chiral phosphorothioate linkages |
topic | Chemical Biology and Nucleic Acid Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4267618/ https://www.ncbi.nlm.nih.gov/pubmed/25398895 http://dx.doi.org/10.1093/nar/gku1115 |
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