Process for the preparation of oligonucleotides

• US 7,635,772 B2
• Filed: 12/16/2003
• Issued: 12/22/2009
• Est. Priority Date: 12/18/2002
• Status: Active Grant

• Current Assignee: Nitto Denko Avecia, Inc. (Nitto Denko Corporation)
• Original Assignee: Avecia Biotechnology Inc (Nitto Denko Corporation)
• Inventors: McCormac, Paul
• Primary Examiner(s): Jiang; Shaojia Anna
• Assistant Examiner(s): OLSON, ERIC
• Application Number:
  US10/539,625
• Publication Number:
  US 20060149052A1
• Time in Patent Office: 2,198 Days
• Field of Search: 424/486, 536/25.4
• US Class Current: 536/25.31
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/14 Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support

Abstract

A process for the synthesis of an oligonucleotide is provided in which an oligonucleotide is assembled on a swellable solid support using the phosphoramidite approach in the presence of an activator, wherein the activator is not tetrazole or a substituted tetrazole. Preferred activators are pyridinium, imidazolinium and benzimidazolinium salts; benzotriazole and derivatives thereof; and saccharin or a saccharin derivative. Preferred swellable solid supports comprise functionalised polystyrene, partially hydrolysed polyvinylacetate or poly(acrylamide).

Claims

1. A process for the synthesis of an oligonucleotide in which an oligonucleotide is assembled on a swellable solid support using the phosphoramidite approach in the presence of a solvent and an activator, wherein, the solvent and swellable support are selected such that a swell ratio of from 5 to 20 is achieved, swell ratio being calculated according to the formula:

2. A process according to claim 1, wherein the activator is the N-methylimidazobe, pyridine or 3-methylpyridine salt of a compound of formula (1) wherein X is O and p is 0.

3. A process according to claim 1, wherein the swellable support comprises functionalised polystyrene, partially hydrolysed polyvinylacetate or poly(acrylamide).

4. A process according to claim 1, wherein the process comprises coupling a nucleoside phosphoramidite with a nucleoside or oligonucleotide comprising a free hydroxy group.

5. A process according to claim 4, wherein the nucleoside phosphoramidite is a deoxyribonucleside-3′

-phosphoramidite or ribonucleside-3′

-phosphoramidite.

6. A process according to claim 4, wherein the nucleoside or oligonucleotide comprising a free hydroxy group comprises a free 5′

-hydroxy group.

7. A process according to claim 4, wherein the nucleoside or oligonucleotide comprising a free hydroxy group is attached to the solid support by a cleavable linker.

8. A process according to claim 1, wherein the solvent is dimethylformamide, N-methylpyrrolidinone, dichloromethane, tetrahydrofuran or pyridine.

9. A process according to claim 1, wherein the assembled oligonucleotide is cleaved from the solid support.

10. A process for the synthesis of an oligonucleotide which comprises coupling a nucleoside phosphoramidite with a nucleoside or oligonucleotide comprising a free hydroxy group in the presence of an activator, wherein:

a) the nucleoside or oligonucleotide comprising a free hydroxy group is attached to a swellable solid support by a cleavable linker, said swellable support being selected from the group consisting of functionalized polystyrene, partially hydrolyzed polyvinylacetate and poly(acrylamide);

b) said activator is a salt formed between an organic base and a compound having the chemical formula;

11. A process according to claim 10, wherein the activator is the N-methylimidazole, pyridine or 3-methylpyridine salt of a compound of formula (1) wherein X is O and p is 0.