Process for the synthesis of pyrazolopyrimidines

• US 20030078413A1
• Filed: 09/12/2001
• Published: 04/24/2003
• Est. Priority Date: 09/12/2001
• Status: Active Grant

• Current Assignee: ELITechGroup Inc. (Bruker Corporation)
• Original Assignee: Epoch Biosciences Inc. (Ngen, Inc.)
• Inventors: Dempcy, Robert O., Reed, Michael W., Adams, A. David
• Granted Patent: US 6,962,991 B2
• Time in Patent Office: Days
• US Class Current: 536/26.1
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• CPC Class Codes: C07H 19/044 Pyrrole radicals

Abstract

The present invention provides a nucleoside comprising a pyrazolopyrimidine base and a process for producing the same. In particular, the processes of the present invention comprises using a halogenated pyrazolopyrimidine base and removing the halogen after the base is coupled to a sugar moiety. The presence of the halogen on the nucleoside base allows facile and economical production of a large quantity of nucleosides.

Claims

1. A PPG phosphoramidite comprising a photolabile hydroxy protecting group, wherein said phosphoramidite nucleoside is of the formula:

2. The PPG phosphoramidite according to claim 1 of the formula:

3. The PPG phosphoramidite according to claim 2, wherein Z³ is —

OR¹³ and Z⁵ is —

OR², where R¹² and R¹³ are those defined in claim 1.

4. The PPG phosphoramidite according to claim 3, wherein the photolabile hydroxy protecting group is selected from the group consisting of α

-methyl-6-nitropiperonyloxycarbonyl, 2-(2-nitrophenyl)-2-methylethoxycarbonyl, 2-(2-nitro-6-chlorophenyl)-2-methylethylsulfonyl, and 3′

,5′

-dimethoxybezoinoxycarbonyl.

5. The PPG phosphoramidite according to claim 4, wherein R¹ and R² together form an amine protecting group.

6. The PPG phosphoramidite according to claim 5, wherein R¹ and R² together form an amine protecting group of the formula:

⊚

CH—

N(CH₃)₂.

7. A process for producing a non-halogenated nucleoside base containing nucleoside comprising:

(a) contacting a halogenated nucleoside base with an activated sugar under conditions sufficient to produce a halogenated nucleoside base containing nucleoside; and

(b) reducing said halogenated nucleoside base containing nucleoside under conditions sufficient to produce said non-halogenated nucleoside base containing nucleoside.

8. The process of claim 7, wherein said non-halogenated nucleoside base containing nucleoside is purified by recrystallization.

9. The process of claim 7, wherein the yield of said non-halogenated nucleoside base containing nucleoside from said halogenated nucleoside base is at least about 50%.

10. The process of claim 7, wherein said halogenated nucleoside base containing nucleoside reducing step comprises hydrogenation of said halogenated nucleoside base containing nucleoside in the presence of a hydrogenation catalyst.

11. The process of claim 7, wherein said non-halogenated nucleoside base containing nucleoside is used in a synthesis of a phosphoramidite nucleoside.

12. The process of claim 11, wherein said phosphoramidite nucleoside is used in a synthesis of an oligonucleoside or an oligonucleotide.

13. A process for producing a nucleoside comprising a hydropyrazolopyrimidine nucleoside base, said process comprising hydrolyzing and reducing or reducing and hydrolyzing an iodopyrazolopyrimidine nucleoside of the formula:

14. The process of claim 13, wherein R¹, R², Y¹, Y², Y⁴, and Y⁶ are hydrogen, and Y³ and Y⁵ are —

OR⁴.

15. The process of claim 14, wherein R⁴ are hydrogen.

16. The process of claim 15 further comprising producing a PPG phosphoramidite of the formula:

17. The process of claim 16, wherein said amine protecting reagent is selected from the group consisting of N,N-dialkylformamide dialkylacetal, and N,N-dialkylacetamide dialkylacetal.

18. The process of claim 16, wherein said hydroxy protecting reagent is a photolabile hydroxy protecting reagent.

19. The process of claim 18, wherein said photolabile hydroxy protecting reagent is selected from the group consisting of 1-(3,4-methylenedioxy-6-nitrophenyl)ethyl chloroformate, 2-(2-nitrophenyl)-2-methylethyl chloroformate, 2-(2-nitro-6-chlorophenyl)-2-methylethylsulfonyl chloride and 3′

,5′

-dimethoxybezoinoxyl chloroformate.

20. The process of claim 16, wherein said hydroxy protecting reagent is an acid labile hydroxy protecting reagent.

21. The process of claim 20, wherein said acid labile hydroxy protecting reagent is selected from the group consisting of trityl halide, monomethoxytrityl halide and dimethoxytrityl halide.

22. The process of claim 16, wherein said activated phosphoramidite is of the formula:

23. The process of claim 22, wherein X² is selected from the group consisting of halide and diisopropylamino.

24. The process of claim 22, wherein R⁹ is dimethoxytrityl and R¹⁰ is a phosphoramidite moiety of the formula —

P[N(i-Pr)₂]OCH₂CH₂CN.

25. The process of claim 13 further comprising producing said nucleoside of Formula I, wherein said nucleoside of Formula I producing step comprises:

contacting an iodopyrazolopyrimidine of the formula;

with an activated sugar of the formula;

under conditions sufficient to produce said nucleoside of Formula I,

wherein

R¹, R², R³, Y¹, Y², Y³, Y⁴, Y⁵, and Y⁶ are those defined claim 13;

and

X¹ is a leaving group.

26. The process of claim 25 further comprising producing said iodopyrazolopyrimidine nucleoside of Formula I from a pyrimidinone of the formula:

27. The process of claim 26, wherein said halogenating agent is selected from the group consisting of POCl₃, iodine monochloride, N-iodosuccinamide and SOCl₂.

28. The process of claim 26, wherein said formylating agent is a compound comprising a formyl group attached to a secondary amino group.

29. The process of claim 28, wherein said formylating agent is selected from the group consisting of dimethyl formamide, 1-formylpiperidine, 1-formylmorpholine and triformamide.

30. The process of claim 26, wherein said iodinating agent is selected from the group consisting of iodine monochloride and N-iodosuccinimide.

31. A process for producing a nucleoside comprising:

(a) contacting an iodopyrazolopyrimidine of the formula;

with an activated sugar of the formula;

under conditions sufficient to produce an deoxy iodopyrazolopyrimidine nucleoside of the formula;

(b) producing an amino dihydro hydropyrazolopyrimidine nucleoside from said deoxy iodopyrazolopyrimidine nucleoside, wherein said amino dihydro hydropyrazolopyrimidine nucleoside is of the formula;

wherein

R³ is alkyl;

R⁵ and R⁶ are hydroxy protecting groups; and

X¹ is a leaving group.

32. The process of claim 31, wherein said step of producing said amino dihydro hydropyrazolopyrimidine nucleoside comprises removing said hydroxy protecting groups R⁵ and R⁶;

hydrolyzing —

OR³ group; and

reducing the iodine.

33. The process of claim 31 further comprising:

(c) contacting said amino dihydro hydropyrazolopyrimidine nucleoside with an amine protecting reagent under conditions sufficient to produce an amine protected nucleoside of the formula;

(d) contacting said amine protected nucleoside with a hydroxy protecting reagent under conditions sufficient to produce an amine/monohydroxy protected nucleoside of the formula;

and

(e) contacting said amine/monohydroxy protected nucleoside with an activated phosphoramidite of the formula;

under conditions sufficient to produce a PPG phosphoramidite of the formula;

wherein

R¹ is hydrogen;

R² is an amine protecting group;

or R¹ and R² together form an amine protecting group;

R⁴ is a hydroxy protecting group; and

X² is a leaving group.

34. The process of claim 33, wherein X² is selected from the group consisting of halide, and —

N(i-Pr)₂.

35. The process of claim 33, wherein R¹ and R² together form a nitrogen protecting group of the formula:

═

CH—

N(CH₃)₂.

36. The process of claim 35, wherein R⁴ is selected from the group consisting of an acid labile hydroxy protecting group and a photolabile hydroxy protecting group.

37. The process of claim 36, wherein R⁴ is selected from the group consisting of dimethoxytrityl, trityl, pixyl, 1,1-bis(4-methoxyphenyl)-1-pyrenylmethyl, (α

-methyl-6-nitropiperonyloxycarbonyl, 2-(2-nitrophenyl)-2-methylethoxycarbonyl, 2-(2-nitro-6-chlorophenyl)-2-methylethylsulfonyl and 3′

,5′

-dimethoxybezoinoxycarbonyl.

38. The process of claim 31, wherein said step (b) comprises reducing the iodide by hydrogenation.

39. The process of claim 31, wherein said iodopyrazolopyrimidine is produced from a pyrimidinone of the formula:

40. The process of claim 39, wherein said halogenating agent is selected from the group consisting of POCl₃, iodine monochloride, N-iodosuccinamide and SOCl₂.

41. The process of claim 40, wherein said halogenating agent is selected from the group consisting of POCl₃ and SOCl₂.

42. The process of claim 39, wherein said formylating agent is selected from the group consisting of dimethyl formamide, 1-formylpiperidine, 1-formylmorpholine and triformamide.

43. The process of claim 39, wherein said iodinating agent is selected from the group consisting of iodine monochloride and N-iodosuccinimide.