Biodegradable lipids for the delivery of active agents

• US 9,012,498 B2
• Filed: 06/03/2011
• Issued: 04/21/2015
• Est. Priority Date: 06/03/2010
• Status: Active Grant

• Current Assignee: Alnylam Pharmaceuticals Incorporated
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Manoharan, Muthiah, Maier, Martin, Jayaraman, Muthusamy, Matsuda, Shigeo, Jayaprakash, Narayanannair K., Rajeev, Kallanthottathil G., Akinc, Akin, Baillie, Thomas A.
• Primary Examiner(s): ZAREK, PAUL E
• Application Number:
  US13/153,200
• Publication Number:
  US 20120027803A1
• Time in Patent Office: 1,418 Days
• Field of Search: 514/512, 514/671, 554/110
• US Class Current: 514/512
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 39/0005 Vertebrate antigens from snakes A61K39/38
• CPC Class Codes: A61K 47/44 Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin synthetic glycerides, e.g. medium-chain triglycerides, A61K47/14
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07C 229/12 to carbon atoms of acyclic carbon skeletons
• 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 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/30 Special therapeutic applications

Abstract

The present invention relates to a cationic lipid having one or more biodegradable groups located in the mid- or distal section of a lipidic moiety (e.g., a hydrophobic chain) of the cationic lipid. These cationic lipids may be incorporated into a lipid particle for delivering an active agent, such as a nucleic acid. The invention also relates to lipid particles comprising a neutral lipid, a lipid capable of reducing aggregation, a cationic lipid of the present invention, and optionally, a sterol. The lipid particle may further include a therapeutic agent such as a nucleic acid.

Claims

1. A compound of formula (I):

2. The compound of claim 1, wherein M¹ and M² are each, independently:

—

OC(O)—

, —

C(O)O—

, —

OC(O)O—

or —

C(O)NH—

.

3. The compound of claim 2, wherein M¹ and M² are each, independently:

—

C(O)O—

or —

OC(O)—

.

4. The compound of claim 2, wherein M¹ and M² are each —

C(O)O—

.

5. The compound of claim 1, wherein R¹ and R² are each alkyl.

6. The compound of claim 5, wherein R¹ and R² are each methyl.

7. The compound of claim 1, wherein each instance of R is, independently, —

CH₂—

, —

C(CH₃)₂—

or —

CH(iPr)-.

8. The compound of claim 1, wherein a is 2, 3, or 4.

9. The compound of claim 1, wherein a carbon atom alpha or beta to a biodegradable group is substituted with one or two alkyl groups.

10. The compound of claim 1, wherein one or more of the following applies:

(i) M¹ and M² are both —

C(O)O—

;

(ii) j and k are both 1;

(iii) j and k are both 0;

(iv) b is 7;

(v) c is 7;

(vi) b and d total 7;

(vii) c and e total 7;

(viii) d and e are each 7;

(ix) n and o are both 1;

(x) f and g are both 0;

(xi) f and h total 1 or 2.

11. The compound of claim 1, selected from:

12. The compound of claim 1, wherein the R¹R²N—

(R)_a—

group is (CH₃)₂N—

(CH₂)₃—

.

13. The compound of claim 1, wherein R¹ and R² are each, independently, C₁-C₄ alkyl.

14. The compound of claim 1, wherein each occurrence of R is, independently, —

CH₂—

or —

CH(CH₃)—

.

15. The compound of claim 1, wherein the sum of b+3j+d+f+3l+h is from about 8 to about 20.

16. The compound of claim 1, wherein the sum of f+3k+e+g+3m+i is from about 8 to about 20.

26. The compound of claim 1, wherein the compound is in the form of a pharmaceutically acceptable salt.

27. The compound of claim 1, wherein the compound is in the form of a cationic lipid.

28. A lipid particle comprising a neutral lipid, a lipid capable of reducing aggregation, and a cationic lipid of claim 27.

29. The lipid particle of claim 28, wherein the neutral lipid is selected from DSPC, DPPC, POPC, DOPE, or SM;

the lipid capable of reducing aggregation is a PEG lipid; and

the lipid particle further comprises a sterol.

30. The lipid particle of claim 28, wherein the cationic lipid is present in a mole percentage of about 20% and about 60%;

the neutral lipid is present in a mole percentage of about 5% to about 25%;

the sterol is present in a mole percentage of about 25% to about 55%; and

the PEG lipid is PEG-DMA, PEG-DMG, or a combination thereof, and is present in a mole percentage of about 0.5% to about 15%.

31. The lipid particle of claim 28, further comprising an active agent.

32. The lipid particle of claim 31, wherein the active agent is a nucleic acid selected from a plasmid, an immunostimulatory oligonucleotide, an siRNA, an antisense oligonucleotide, a microRNA, an antagomir, an aptamer, and a ribozyme.

33. The lipid particle of claim 28, wherein the lipid particle has an in vivo half life (t_1/2) of less than about 3 hours.

34. The lipid particle of claim 28, wherein the lipid particle has an in vivo half life (t_1/2) of less than about 10% of that for a lipid particle containing the same cationic lipid without a biodegrable group.

35. A pharmaceutical composition comprising a lipid particle of claim 28 and a pharmaceutically acceptable carrier.

36. A method of modulating the expression of a target gene in a cell, comprising providing to the cell a lipid particle of claim 28.

37. The method of claim 26, wherein the active agent is a nucleic acid selected from a plasmid, an immunostimulatory oligonucleotide, an siRNA, an antisense oligonucleotide, a microRNA, an antagomir, an aptamer, and a ribozyme.

38. A method of inducing an immune response in a subject, comprising providing to the subject the pharmaceutical composition of claim 35, wherein the active agent is an immunostimulatory oligonucleotide.

39. The method of claim 38, wherein the target gene is selected from the group consisting of Factor VII, Eg5, PCSK9, TPX2, apoB, SAA, TTR, RSV, PDGF beta gene, Erb-B gene, Src gene, CRK gene, GRB2 gene, RAS gene, MEKK gene, JNK gene, RAF gene, Erk1/2 gene, PCNA(p21) gene, MYB gene, JUN gene, FOS gene, BCL-2 gene, Cyclin D gene, VEGF gene, EGFR gene, Cyclin A gene, Cyclin E gene, WNT-1 gene, beta-catenin gene, c-MET gene, PKC gene, NFKB gene, STAT3 gene, survivin gene, Her2/Neu gene, SORT1 gene, XBP1 gene, topoisomerase I gene, topoisomerase II alpha gene, p73 gene, p21(WAF1/CIP1) gene, p27(KIP1) gene, PPM1D gene, RAS gene, caveolin I gene, MIB I gene, MTAI gene, M68 gene, tumor suppressor genes, and p53 tumor suppressor gene.

40. The method of claim 39, wherein the target gene contains one or more mutations.

41. A method of delivering a nucleic acid molecule comprising administering a lipid particle of claim 32 to a subject.

17. A compound of formula TB

18. The compound of claim 17, wherein R⁹ and R¹⁰ are each independently C₄-C₁₂ alkylene or C₄-C₁₂ alkenylene, M¹ and M² are —

C(O)O—

, and R¹¹ and R¹² are C₄-C₁₂ alkylene or C₄-C₁₂ alkenylene.

19. The compound of claim 17, wherein R⁹, M¹, and R¹¹ are together at 12 to 24 carbons atoms in length.

20. The compound of claim 17, wherein, R¹⁰, M², and R¹² are together at 12 to 24 carbons atoms in length.

21. The compound of claim 17, wherein the R¹R²N—

(R)_a—

group is (CH₃)₂N—

(CH₂)₃—

.

22. A compound of formula IC

23. The compound of claim 22, wherein each group M is —

COOR¹³ wherein each R¹³ is, independently, alkyl.

24. The compound of claim 22, wherein the R¹R²N—

(R)_a—

group is (CH₃)₂N—

(CH₂)₃—

.

25. A compound selected from a compound of formulas II-XXIII: