Oligonucleotide synthesis with alternative solvents

Details Oligonucleotide synthesis with alternative solvents Oligonucleotide synthesis with alternative solvents

C - Chemistry, Metallurgy

07

H

C07H 21/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01)

Patent

CA 2540692

The invention provides for methods of manufacturing an oligonucleotide comprising a pentavalent phosphate triester. In particular, the method comprises providing a 5' blocked-nucleoside, deblocking the 5' blocked- nucleoside to form a 5' OH-nucleoside, coupling the 5' OH-nucleoside with a phosphoramidite to form and oligonucleotide comprising a trivalent phosphite triester; and oxidizing the oligonucleotide comprising a trivalent phosphite triester to the oligonucleotide comprising a pentavalent phosphate triester. In some embodiments, the wash between any of the steps above is with at least one solvent wash comprising a toluene.

L'invention concerne des procédés de fabrication d'un oligonucléotide comprenant un triester de phosphate pentavalent. Plus précisément, le procédé consiste à utiliser un nucléoside bloqué en 5', à débloquer celui-ci de manière à former un nucléoside à extrémité 5' OH, à coupler celui-ci à une phosphoramidite afin de former un oligonucléotide comprenant un triester de phosphite trivalent; et à oxyder l'oligonucléotide comprenant un triester de phosphite trivalent à l'oligonucléotide comprenant un triester de phosphate pentavalent. Dans quelques modes de réalisation, le lavage entre les étapes susmentionnées est effectué au moyen d'au moins d'un solvant comprenant un toluène.

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