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Updated: Jul 5, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Chemical phosphorylation of deoxyribonucleosides and thermolytic DNA oligonucleotides
Cristina Ausín1, Andrzej Grajkowski, Jacek Cieślak
1Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA.
Abstract:
The phosphorylating reagent bis[S-(4,4'-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite is prepared in three steps from commercial methyl thioglycolate and diisopropylphosphoramidous dichloride. The phosphorylating reagent has been used successfully in the solid-phase synthesis of deoxyribonucleoside 5'-/3'-phosphate or -thiophosphate monoesters and oligonucleotide 5'-phosphate/-thiophosphate monoesters. Bis[S-(4,4'-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite has also been employed in the construction of a thermolytic dinucleotide prodrug model to evaluate the ability of the reagent to produce thermosentive oligonucleotide prodrugs under mild temperature conditions ( approximately 25 degrees C) for potential therapeutic applications.
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