COMPOSITIONS AND METHODS FOR PHOSPHORAMIDITE AND OLIGONUCLEOTIDE SYNTHESIS

• US 20190077817A1
• Filed: 03/13/2017
• Published: 03/14/2019
• Est. Priority Date: 03/13/2016
• Status: Active Grant

• Current Assignee: WAVE Life Sciences Ltd.
• Original Assignee: WAVE Life Sciences Ltd.
• Inventors: Butler, David Charles Donnell, Marappan, Subramanian
• Granted Patent: US 11,407,775 B2
• Time in Patent Office: Days
• CPC Class Codes: C07F 9/65844 the phosphorus atom being part of a five-membered ring which may be condensed with another ring system
• CPC Class Codes: C07H 1/06 Separation; Purification
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/16 Purine radicals

Abstract

The present disclosure, among other things, provides technologies for preparing and purifying phosphoramidites for oligonucleotide synthesis.

Claims

1. A method for purifying a phosphoramidite, comprising steps of:

a) removing water from a purification medium;

b) contacting the purification medium with a phosphoramidite; and

c) optionally using the purification medium to purify a phosphoramidite.

5. The method of any one of the preceding claims, wherein the step of removing water from the purification medium comprises heating the purification medium.

6. The method of any one of the claims 1-4, wherein the step of removing water from the purification medium comprises contacting the purification medium with a hygroscopic solvent system.

7. The method of claim 6, wherein the hygroscopic solvent system consists of one hygroscopic solvent.

8. The method of claim 7, wherein the hygroscopic solvent is selected from acetaldehyde, acetic acid, acetone, acetonitrile, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2-butoxyethanol, butyric acid, diethanolamine, diethylenetriamine, dimethyl carbonate, dimethylformamide, dimethoxyethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethylamine, ethylene glycol, formic acid, furfuryl alcohol, glycerol, methanol, methyl diethanolamine, methyl isocyanide, 1-propanol, 1,3-propanediol, 1,5-pentanediol, 2-propanol, propanoic acid, propylene glycol, pyridine, tetrahydrofuran, and triethylene glycol.

9. The method of claim 8, wherein the hygroscopic solvent is selected from methanol, acetonitrile, and ethyl acetate, optionally with triethylamine up to 50%.

10. The method of claim 8, wherein the hygroscopic solvent is methanol.

15. The method of any one of the preceding claims, comprising chromatography purification of the phosphoramidite using the purification medium at a temperature higher and/or lower than room temperature.

16. A composition comprising:

a) a purification medium pre-treated by step a) of any one of the preceding claims;

b) a phosphoramidite.

18. The method or composition of any one of the preceding claims, wherein the phosphoramidite has the structure of formula II.

2. A method for improving recovery rate of a phosphoramidite, comprising steps of:

a) removing water from a purification medium;

b) adding the phosphoramidite to the purification medium; and

c) eluting the phosphoramidite from the purification medium with a solvent system;

wherein the recovery rate is higher than a reference recovery rate when step a) is absent.

3. The method of claim 2, wherein the difference between the improved recovery rate and the reference recovery rate is at least about 10%, 20%, 30%, 40% or 50%.

4. A method for decreasing decomposition of a phosphoramidite when the phosphoramidite contacts a purification medium, comprising:

a) removing water from the purification medium.

11. A method comprising steps of:

a) pre-treating a purification medium;

b) contacting the purification medium with a phosphoramidite; and

b) optionally using the purification medium to purify a phosphoramidite.

12. The method of claim 11, wherein the pre-treatment comprises heating the purification medium.

13. The method of claim 11, wherein the pre-treatment comprises contacting the purification medium with a first solvent system comprises a hygroscopic solvent.

14. The method of any one of claims 11-13, wherein the purification medium is silica gel.

17. A composition comprising:

a) silica gel pre-treated with a hygroscopic solvent;

b) a phosphoramidite.

19. A method or a composition of any one of embodiments 1-144.