Process for purifying oligonucleotide synthons
View Patent ↗A process for the purification of an oligonucleotide synthon is provided. The process comprises subjecting an organic solution comprising an oligonucleotide synthon and lower molecular weight impurities to nanofiltration whereby the ratio of an oligonucleotide synthon to lower molecular weight impurities in the solution is increased after the nanofiltration. Preferably, the oligonucleotide synthon is a nucleoside phosphoramidite or nucleoside H-phosphonate. The nanofiltration membrane is preferably a polyimide membrane having a molecular weight cut off of 400.
1. A process for the purification of a nucleoside phosphoramidite or nucleoside H-phosphonate oligonucleotide synthon from an organic solution of a nucleoside phosphoramidite or nucleoside H-phosphonate oligonucleotide synthon and lower molecular weight impurities, which comprises subjecting said organic solution to nanofiltration whereby the ratio of synthon to lower molecular weight impurities in the solution is increased after the nanofiltration.
2. A process according to claim 1 , wherein the oligonucleotide synthon is a compound of formula (1):
wherein R 1 is a protecting group, B is a nucleoside base, R 2 represents —H, —F, —OR 4 , —NR 5 R 6 , —SR 7 , or aliphatic group, each R 3 independently is a C 1-6 alkyl group, PG is a phosphorus protecting group, R 4 represents —H, an aliphatic group, an aryl group, an aralkyl, an alcohol protecting group, or —(CH 2 ) q —NR 9 R 10 , R 5 and R 6 are each, independently, —H, an aliphatic group, or an amine protecting group, or R 5 and R 6 taken together with the nitrogen to which they are attached are a heterocyclyl group, R 7 represents —H, an aliphatic group, or a thiol protecting group, R 9 and R 10 are each, independently, —H, an aryl group, a heteroaryl group, an aliphatic group, an aralkyl group, a heteroaralkyl group or an amine protecting group, or R 9 and R 10 taken together with the nitrogen to which they are attached form a heterocyclyl group; and q is an integer from 1 to about 6.
3. A process according to claim 2 , wherein PG is a betacyanoethyl group, and each R 3 is an isopropyl group.
4. A process according to claim 1 , wherein a polyimide nanofiltration membrane is employed.
5. A process according to claim 1 , wherein a nanofiltration membrane having a molecular weight cut off of 400 is employed.
6. A process according to claim 1 , wherein the process is operated in cross flow configuration.
7. A process according to claim 1 , wherein the process employs a pressure of from 15 to 35 bar.
8. A process according to claim 1 , wherein fresh organic solvent corresponding to the volume passed through the nanofiltration membrane is added into the retained synthon solution.
9. A process according to any one of claims 1 to 3 , wherein a polyimide nanofiltration membrane having a molecular weight cut-off of 400 is employed.
10. A process according to claim 9 , wherein the process is operated in cross flow configuration.
11. A process according to any one of claims 1 to 3 , wherein a polyimide nanofiltration membrane is employed and fresh organic solvent corresponding to the volume passed through the nanofiltration membrane is added into the retained synthon solution.
12. A process according to claim 11 , wherein a polyimide nanofiltration membrane having a molecular weight cut-off of 400 is employed.
13. A process according to claim 12 , wherein the process is operated in cross flow configuration.