Klabenkova, K.V.; Zhdanova, P.V.; Burakova, E.A.; Bizyaev, S.N.; Fokina, A.A.; Stetsenko, D.A. A Convenient Oligonucleotide Conjugation via Tandem Staudinger Reaction and Amide Bond Formation at the Internucleotidic Phosphate Position. Int. J. Mol. Sci.2024, 25, 2007.
Klabenkova, K.V.; Zhdanova, P.V.; Burakova, E.A.; Bizyaev, S.N.; Fokina, A.A.; Stetsenko, D.A. A Convenient Oligonucleotide Conjugation via Tandem Staudinger Reaction and Amide Bond Formation at the Internucleotidic Phosphate Position. Int. J. Mol. Sci. 2024, 25, 2007.
Klabenkova, K.V.; Zhdanova, P.V.; Burakova, E.A.; Bizyaev, S.N.; Fokina, A.A.; Stetsenko, D.A. A Convenient Oligonucleotide Conjugation via Tandem Staudinger Reaction and Amide Bond Formation at the Internucleotidic Phosphate Position. Int. J. Mol. Sci.2024, 25, 2007.
Klabenkova, K.V.; Zhdanova, P.V.; Burakova, E.A.; Bizyaev, S.N.; Fokina, A.A.; Stetsenko, D.A. A Convenient Oligonucleotide Conjugation via Tandem Staudinger Reaction and Amide Bond Formation at the Internucleotidic Phosphate Position. Int. J. Mol. Sci. 2024, 25, 2007.
Abstract
A Staudinger reaction on solid phase between an electronodeficit organic azide such as sulfonyl azide, and the phosphite triester formed upon phosphoramidite coupling is a convenient method for chemical modification of oligonucleotides at the internucleotidic phosphate position. In this work, 4-carboxybenzenesulfonyl azide either with a free carboxy group or in the form of an activated ester such as pentafluorophenyl, 4-nitrophenyl, or pentafluorobenzyl ester were used to introduce a carboxylic acid function to the terminal or internal internucleotidic phosphate of an oligonucleotide via Staudinger reaction. A subsequent treatment with excess primary alkyl amine followed by usual work-up, after prior activation with a suitable peptide coupling agent such as HBTU/HOBt in the case of a free carboxyl, afforded amide-linked oligonucleotide conjugates in good yields including multiple conjugations of up to the exhaustive modification at each phosphate position for a weakly activated pentafluorobenzyl ester, whereas more strongly activated and, thus, more reactive aryl esters provided only single conjugations at the 5′-end. The conjugates synthesized include those with di- and polyamines that introduce a positively charged side-chain to potentially assist intracellular delivery of the oligonucleotide.
Keywords
nucleic acid; antisense oligonucleotide; conjugation; sulfonyl azide; pentafluorophenyl and 4-nitrophenyl active esters; carboxylic acid group modification; pentafluorobenzyl
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
