Process for triphosphate oligonucleotide synthesis

US9035041B2 · kind B2 · utility

Assignee

• Alnylam Pharmaceuticals, Inc. · US

Inventors

• Ivan Zlatev · Cambridge, US
• François Morvan · Castelnau-le-Lez, FR
• Jean-Jacques Vasseur · Juvignac, FR
• Francoise Debart · Juvignac, FR
• Muthiah Manoharan · Carlsbad, US

Key dates

• Filing date: Dec 22, 2009
• Grant date: May 19, 2015
• Expiry date: Aug 31, 2030

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2310/321
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

This invention relates to a process for preparing an oligonucleotide 5′-triphosphate. The process comprises the steps of: (a) synthesizing an oligonucleotide having a 5′ hydroxyl moiety; (b) reacting the 5′ hydroxyl moiety with a reagent of formula I:to convert the 5′ hydroxyl moiety to a 5′-H-phosphonate, wherein R1 and R2 are each independently selected from the group consisting of haloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, and substituted heterocycle, acyl, phosphoryl, substituted alkyl acyl, substituted heteroalkyl acyl, substituted aryl acyl or substituted heteroaryl acyl, substituted alkyl phosphoryl, substituted heteroalkyl acyl, substituted aryl phosphoryl, and substituted heteroaryl phosphoryl; (c) activating the H-phosphonate of step (b) by reacting the H-phosphonate with a silylating agent, a halogenated oxidizing agent, a nitrogen-containing heteroaryl, or a combination thereof, to form an activated H-phosphonate; and (d) treating the oligonucleotide having an activated H-phosphonate from step (c) with a poly(alkylammonium)pyrophosphate.