Nucleoside phosphorothioate derivatives, synthesis and use thereof

• US 5,708,161 A
• Filed: 01/23/1996
• Issued: 01/13/1998
• Est. Priority Date: 01/25/1995
• Status: Expired due to Fees

• Current Assignee: Kings College London (University of London)
• Original Assignee: Kings College London (University of London)
• Inventors: Reese, Colin Bernard
• Primary Examiner(s): Kunz, Gary L.
• Application Number:
  US08/590,280
• Time in Patent Office: 721 Days
• Field of Search: 536/25.3, 536/25.33
• US Class Current: 536/25.33
• CPC Class Codes: C07D 207/48 Sulfur atoms
• CPC Class Codes: C07D 209/48 with oxygen atoms in positions 1 and 3, e.g. phthalimide
• 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: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

Inter alia, a process for the production of a nucleoside phosphorothioate derivative corresponding to the following general formula (I): ##STR1## characterized in that it comprises reacting a salt of a nucleoside H-phosphonate corresponding to the following general formula (II): ##STR2## with a thiol transfer agent corresponding to the following general formula (III):

R.sup.2 --S--X                                             (III)

in the presence of a silylating agent and a base in a suitable solvent; and working-up the reaction mixture to isolate the desired product and change the counter-cation as desired;

wherein

M⁺ represents a suitable counter-cation;

R¹ represents an appropriate nucleoside or nucleoside analogue;

R² represents a desired protecting group, which may ultimately be removable by cleavage of its bond to the sulfur atom; and

X represents a leaving group,

is disclosed.

Claims

1. A process for the production of a nucleoside phosphorothioate derivative corresponding to the following general formula (I):

##STR15## wherein it comprises reacting a salt of a nucleoside H-phosphonate corresponding to the following general formula (II);

##STR16## with a thiol transfer agent corresponding to the following general formula (III);

space="preserve" listing-type="equation">R.sup.2 --S--X (III)

in the presence of a silylating agent and a base in a suitable solvent; and

working-up the reaction mixture to isolate the desired product and change the counter-cation as desired;

wherein

M⁺ represents a suitable counter-cation;

R¹ represents an appropriate nucleoside or nucleoside analogue;

R² represents a desired protecting group, which is optionally ultimately removed by cleavage of its bond to the sulfur atom; and

X represents a leaving group.

2. A process as claimed in claim 1 wherein R² represents alkyl, substituted alkyl, aralkyl, substituted aralkyl, alkenyl or substituted alkenyl.

3. A process as claimed in claim 2 wherein R² represents methyl, 2-cyanoethyl, benzyl, mononitrobenzyl, dinitrobenzyl or allyl.

4. A process as claimed in claim 1 wherein X represents the conjugate base of succinimide or phthalimide, either of which is optionally substituted.

5. A process as claimed in claim 2 wherein X represents the conjugate base of succinimide or phthalimide, either of which is optionally substituted.

6. A process as claimed in claim 3 wherein X represents the conjugate base of succinimide or phthalimide, either of which is optionally substituted.

7. A process as claimed in claim 1 wherein R² is not removed.

8. A process as claimed in claim 2 wherein R² is not removed.

9. A process as claimed in claim 3 wherein R² is not removed.

10. A process as claimed in claim 4 wherein R² is not removed.

11. A process as claimed in claim 5 wherein R² is not removed.

12. A process as claimed in claim 6 wherein R² is not removed.

13. The method of claim 1 wherein the thiol transfer agent is N-(2-cyanoethylmercapto)-phthalimide or N-(2-cyanoethylmercapto)-succinimide.

14. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 2 is used in a phosphotriester method.

15. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 3 is used in a phosphotriester method.

16. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 4 is used in a phosphotriester method.

17. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 5 is used in a phosphotriester method.

18. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 6 is used in a phosphotriester method.

19. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 7 is used in a phosphotriester method.

20. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 8 is used in a phosphotriester method.

21. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 9 is used in a phosphotriester method.

22. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 10 is used in a phosphotriester method.

23. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 11 is used in a phosphotriester method.

24. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 11 is used in a phosphotriester method.

25. A process for the production of an oligonucleotide phosphorothioate analogue wherein the nucleoside phosphorothioate derivative (I) produced by the process of claim 12 is used in a phosphotriester method.