Cargando…
Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents
Chemically-modified derivatives of cytidine, bearing a 5-(N-substituted-carboxamide) functional group, are new reagents for use in aptamer discovery via the SELEX process (Systematic Evolution of Ligands by EXponential enrichment). Herein, we disclose a practical synthesis of 5-(N-benzylcarboxamide)...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353258/ https://www.ncbi.nlm.nih.gov/pubmed/25710355 http://dx.doi.org/10.1080/15257770.2014.978011 |
_version_ | 1782360582204162048 |
---|---|
author | Rohloff, John C. Fowler, Catherine Ream, Brian Carter, Jeffrey D. Wardle, Greg Fitzwater, Tim |
author_facet | Rohloff, John C. Fowler, Catherine Ream, Brian Carter, Jeffrey D. Wardle, Greg Fitzwater, Tim |
author_sort | Rohloff, John C. |
collection | PubMed |
description | Chemically-modified derivatives of cytidine, bearing a 5-(N-substituted-carboxamide) functional group, are new reagents for use in aptamer discovery via the SELEX process (Systematic Evolution of Ligands by EXponential enrichment). Herein, we disclose a practical synthesis of 5-(N-benzylcarboxamide)-2′-deoxycytidine, and the corresponding 5-(N-1-naphthylmethylcarboxamide)- and 5-(N-3-phenylpropylcarboxamide)-2′-deoxycytidine analogs, as both the suitably-protected 3′-O-cyanoethylphosphoramidite reagents (CEP; gram scale) and the 5′-O-triphosphate reagents (TPP; milligram-scale). The key step in the syntheses is a mild, palladium(0)-catalyzed carboxyamidation of an unprotected 5-iodo-cytidine. Use of the CEP reagents for solid-phase oligonucleotide synthesis was demonstrated and incorporation of the TPP reagents by KOD polymerase in a primer extension assay confirmed the utility of these reagents for SELEX. Finally, the carboxyamidation reaction was also used to prepare the nuclease-resistant sugar-variants: 5-(N-benzylcarboxamide)-2′-O-methyl-cytidine and 5-(N-3-phenylpropylcarboxamide)-2′-deoxy-2′-fluoro-cytidine. |
format | Online Article Text |
id | pubmed-4353258 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-43532582015-03-23 Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents Rohloff, John C. Fowler, Catherine Ream, Brian Carter, Jeffrey D. Wardle, Greg Fitzwater, Tim Nucleosides Nucleotides Nucleic Acids Original Articles Chemically-modified derivatives of cytidine, bearing a 5-(N-substituted-carboxamide) functional group, are new reagents for use in aptamer discovery via the SELEX process (Systematic Evolution of Ligands by EXponential enrichment). Herein, we disclose a practical synthesis of 5-(N-benzylcarboxamide)-2′-deoxycytidine, and the corresponding 5-(N-1-naphthylmethylcarboxamide)- and 5-(N-3-phenylpropylcarboxamide)-2′-deoxycytidine analogs, as both the suitably-protected 3′-O-cyanoethylphosphoramidite reagents (CEP; gram scale) and the 5′-O-triphosphate reagents (TPP; milligram-scale). The key step in the syntheses is a mild, palladium(0)-catalyzed carboxyamidation of an unprotected 5-iodo-cytidine. Use of the CEP reagents for solid-phase oligonucleotide synthesis was demonstrated and incorporation of the TPP reagents by KOD polymerase in a primer extension assay confirmed the utility of these reagents for SELEX. Finally, the carboxyamidation reaction was also used to prepare the nuclease-resistant sugar-variants: 5-(N-benzylcarboxamide)-2′-O-methyl-cytidine and 5-(N-3-phenylpropylcarboxamide)-2′-deoxy-2′-fluoro-cytidine. Taylor & Francis 2015-03-04 2015-02-24 /pmc/articles/PMC4353258/ /pubmed/25710355 http://dx.doi.org/10.1080/15257770.2014.978011 Text en © 2015 Specialty Chemistry. Published with license by Taylor & Francis This is an Open Access article. Non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly attributed, cited, and is not altered, transformed, or built upon in any way, is permitted. The moral rights of the named author(s) have been asserted. |
spellingShingle | Original Articles Rohloff, John C. Fowler, Catherine Ream, Brian Carter, Jeffrey D. Wardle, Greg Fitzwater, Tim Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title | Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title_full | Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title_fullStr | Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title_full_unstemmed | Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title_short | Practical Synthesis of Cytidine-5-Carboxamide-Modified Nucleotide Reagents |
title_sort | practical synthesis of cytidine-5-carboxamide-modified nucleotide reagents |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353258/ https://www.ncbi.nlm.nih.gov/pubmed/25710355 http://dx.doi.org/10.1080/15257770.2014.978011 |
work_keys_str_mv | AT rohloffjohnc practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents AT fowlercatherine practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents AT reambrian practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents AT carterjeffreyd practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents AT wardlegreg practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents AT fitzwatertim practicalsynthesisofcytidine5carboxamidemodifiednucleotidereagents |