The mechanism of the phosphoramidite synthesis of polynucleotides

Mark A. Russell, Andrew P. Laws, John H. Atherton, Michael I. Page

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The mechanism of the coupling step in polynucleotide synthesis using 5′-4,4′-dimethoxytritylthymidine-3′-β-cyanoethyl-N, N-diisopropylphosphoramidite as the phosphitylating agent and catalysed by the salt of saccharin and N-methylimidazole in acetonitrile has been studied by 31P NMR. Pre- and post-equilibria between the activator salt and released diisopropylamine have been examined by 1H NMR and ITC, which show that the salt between saccharin and diisopropylamine will be present in acetonitrile. Activation of the phosphoramidite by the salt of saccharin and N-methylimidazole involves nucleophilic catalysis and the formation of a reactive saccharin adduct bonded through its carbonyl oxygen to phosphorus. The rate constants for the reaction of the 4-methoxyphenol with 5′-4,4′- dimethoxytritylthymidine-3′-β-cyanoethyl-N,N- diisopropylphosphoramidite in the presence of saccharin-N-methylimidazole salt show a non-linear dependence on phenol concentration, becoming independent at high phenol concentrations, compatible with a change in rate limiting step from the alcoholysis step to the activation step.

Original languageEnglish
Pages (from-to)3270-3275
Number of pages6
JournalOrganic and Biomolecular Chemistry
Volume6
Issue number18
DOIs
Publication statusPublished - 29 Jul 2008

Fingerprint

polynucleotides
Saccharin
Polynucleotides
Salts
salts
synthesis
Phenol
phenols
acetonitrile
Chemical activation
Nuclear magnetic resonance
activation
nuclear magnetic resonance
Catalysis
Phosphorus
adducts
catalysis
phosphorus
Rate constants
phosphoramidite

Cite this

Russell, Mark A. ; Laws, Andrew P. ; Atherton, John H. ; Page, Michael I. / The mechanism of the phosphoramidite synthesis of polynucleotides. In: Organic and Biomolecular Chemistry. 2008 ; Vol. 6, No. 18. pp. 3270-3275.
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The mechanism of the phosphoramidite synthesis of polynucleotides. / Russell, Mark A.; Laws, Andrew P.; Atherton, John H.; Page, Michael I.

In: Organic and Biomolecular Chemistry, Vol. 6, No. 18, 29.07.2008, p. 3270-3275.

Research output: Contribution to journalArticle

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