Modified iRNA agents

• US 7,745,608 B2
• Filed: 08/10/2004
• Issued: 06/29/2010
• Est. Priority Date: 04/17/2003
• Status: Active Grant

• Current Assignee: Alnylam Pharmaceuticals Incorporated
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Manoharan, Muthiah, Rajeev, Kallanthottathil G., Kesavan, Venkitasamy
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US10/916,185
• Publication Number:
  US 20050107325A1
• Time in Patent Office: 2,149 Days
• Field of Search: None
• US Class Current: 536/24.5
• CPC Class Codes: A61K 47/55 the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/323 modified ring structure
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance
• CPC Class Codes: C12N 2330/30 chemically synthesised

Abstract

The invention relates to iRNA agents, which preferably include a monomer in which the ribose moiety has been replaced by a moiety other than ribose. The inclusion of such a monomer can allow for modulation of a property of the iRNA agent into which it is incorporated, e.g., by using the non-ribose moiety as a point to which a ligand or other entity, e.g., a lipophilic moiety. e.g., cholesterol, is directly, or indirectly, tethered. The invention also relates to methods of making and using such modified iRNA agents.

Claims

1. An iRNA agent comprising a first strand and a second strand, wherein at least one subunit having a formula (I) is incorporated into at least one of said strands, and at least one of said strands leads to the cleavage of a complementary target sequence via a RISC dependent mechanism:

2. The compound of claim 1, wherein R³ is OR^a.

3. The compound of claim 2, wherein

4. The compound of claim 2, wherein A is O.

5. The compound of claim 2, wherein A is S.

6. The compound of claim 1, wherein R³ is OR^b.

7. The compound of claim 2, wherein R⁷ is (CH₂)₅NHR^d or (CH₂)₅NHC(O)R^d.

8. The compound of claim 2, wherein R^d is a cholesterol radical.

9. The compound of claim 1, wherein R⁹ is OR^a.

10. The compound of claim 9, wherein

11. The compound of claim 1, wherein R⁹ is OR^b.

12. The compound of claim 1, wherein one of R³ and R⁹ is OR^a.

13. The compound of claim 1, wherein one of R³ and R⁹ is OR^b.

14. A kit comprising an iRNA agent of claim 1, a sterile container in which the iRNA agent is enclosed, and instructions for use.

15. The compound of claim 1, wherein one of R³ or R⁹ is OH.

21. The compound of claim 1, wherein R³ is H, R⁵ is H, and R⁹ is OH.

22. The compound of claim 1, wherein X is N(CO)R⁷.

23. The compound of claim 22, wherein R⁷ is a C₁-C₂₀ alkyl substituted with NHC(O)R^d.

24. The compound of claim 23, wherein R⁷ is a C₃-C₈ alkyl substituted with NHC(O)R^d.

25. The compound of claim 24, wherein R⁷ is a C₅-C₇ alkyl substituted with NHC(O)R^d.

26. The compound of claim 23, wherein R⁷ is a C₅ alkyl substituted with NHC(O)R^d.

27. The compound of claim 26, wherein R^d is a cholesterol radical.

28. The compound of claim 27, wherein R⁹ is OH.

29. The compound of claim 1, wherein R^d is a cholesterol radical.

30. The agent of claim 1, wherein R^d is uvaol.

31. The agent of claim 1, wherein R^d is hecogenin.

32. The agent of claim 1, wherein R^d is diosgenin.

33. The agent of claim 1, wherein R^d is cholic acid.

34. The agent of claim 1, wherein R^d is dihydrotestosterone.

35. The agent of claim 1, wherein R^d is a cholesteryl residue.

36. The agent of claim 1, wherein R^d is an O3-(oleoyl)lithocholic acid residue.

37. The agent of claim 1, wherein R^d is an O3-(oleoyl) cholenic acid residue.

38. The agent of claim 1, wherein A is O in each occurrence.

39. The agent of claim 1, wherein A is S in at least one occurrence.

40. The agent of claim 1, wherein formula (I) is incorporated into a sense strand.

41. The agent of claim 40, wherein formula (I) is incorporated into the 3′

end of the sense strand.

42. The agent of claim 1, wherein formula (I) is incorporated into an antisense strand.

43. The agent of claim 42, wherein formula (I) is incorporated into the 3′

end of the antisense strand.

16. An iRNA agent comprising a first strand and a second strand, wherein at least one subunit having a formula (I′

) is incorporated into at least one of said strands;

17. The compound of claim 16, wherein R⁹ is OH.

18. The compound of claim 16, wherein R⁹ is OR^a.

19. The compound of claim 16, wherein R⁹ is OR^b.

20. The compound of claim 16, wherein