LIPID FORMULATION

• US 20120101148A1
• Filed: 01/29/2010
• Published: 04/26/2012
• Est. Priority Date: 01/29/2009
• Status: Abandoned Application

• Current Assignee: Arbutus Biopharma Corporation
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Querbes, William, Qin, Xiaojun, Cantley, William, Borodovsky, Anna, Manoharan, Muthiah, Jayaraman, Muthusamy, Rajeev, Kallanthottathil G., Aking, Akin, Wong, Frances M.P., Dorkin, Joseph R., De, Soma
• Application Number:
  US13/147,189
• Publication Number:
  US 20120101148A1
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 48/0008 characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

The invention features an improved lipid formulation comprising a cationic lipid of formula (A), a neutral lipid, a sterol and a PEG or PEG-modified lipid, where R₁ and R₂ are independently alkyl, alkenyl or alkynyl, each can be optionally substituted, and R₃ and R₄ are independently lower alkyl or R₃ and R₄ can be taken together to form an optionally substituted heterocyclic ring. In one embodiment, R₁ and R₂ are independently selected from oleoyl, pamitoyl, steroyl, linoleyl and R₃ and R₄ are methyl. Also disclosed are targeting lipids, and specific lipid formulations comprising such targeting lipids.

Claims

1. A lipid formulation comprising 45-65% of cationic lipid of formula A, 5-10% of the neutral lipid, 25-40% of the sterol, and 0.5-5% of the PEG or PEG-modified lipid, wherein formula A is

2. The lipid formulation of claim 1, wherein the neutral lipid is selected from DSPC, DPPC, DMPC, DPPC, POPC, DOPE and SM.

3. The lipd formulation of claim 1, wherein the sterol is cholesterol.

4. The lipid formulation of claim 1, wherein the PEG lipid is PEG-C₁₄ to PEG-C₂₂, PEG-Cer₁₄ to PEG-C₂₀, or PEG-DSPE.

5. The lipid formulation of claim 1, wherein R₁ and R₂ of formula A are selected from selected from oleoyl, pamitoyl, steroyl, linoleyl and R₃ and R₄ are methyl.

6. The lipid formulation of claim 1, wherein the cationic lipid of formula A is 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, the neutral lipid is DSPC, the sterol is cholesterol and the PEG lipid is PEG-DMG.

7. The lipid formulation of claim 6 comprising about 60% of cationic lipid of formula A, about 7.5% of the neutral lipid, about 31% of the sterol, and about 1.5% of the PEG or PEG-modified lipid.

8. The lipid formulation of claim 7, wherein the formulation is prepared by an in-line mixing method.

9. The lipid formulation of claim 6 comprising about 57.5% of cationic lipid of formula A, about 7.5% of the neutral lipid, about 31.5% of the sterol, and about 3.5% of the PEG or PEG-modified lipid.

10. The lipid formulation of claim 9, wherein the formulation is prepared by an extrusion method.

11. The lipid formulation of claim 1, further comprising a therapeutic agent.

12. The lipid formulation of claim 11, wherein the therapeutic agent comprises a nucleic acid.

13. The lipid formulation of claim 11, wherein the nucleic acid is selected from antisense, siRNA, ribozyme and microRNA.

14. The lipid formulation of claim 12, wherein the ratio of lipid:

nucleic acid is about 3;

1 to about 15;

1.

15. The lipid formulation of claim 14, wherein the ratio of lipid:

nucleic acid is about 5;

1 to about 13;

1.

16. The lipid formulation of claim 15, wherein the ratio of lipid nucleic acid is about 7:

1 to about 11;

1

17. The lipid formulation of claim 1, further comprising at least one apolipoprotein.

18. The lipid formulation of claim 17, wherein the apolipotprotein is selected from the group consisting of ApoA-I, ApoA-II, ApoA-IV, ApoA-V and ApoE, active polymorphic forms, isoforms, variants and mutants, and fragments or truncated forms thereof.

19. The lipid formulation of claim 17, wherein the apolipotprotein is ApoE, active polymorphic forms, isoforms, variants and mutants, and fragments or truncated forms thereof.

20. The lipid formulation of claim 1, further comprising a targeting lipid.

21. The lipid formulation of claim 1, further comprising a targeting lipid comprising N-acetyl galactosamine as a targeting moiety.

22. The formulation of claim 21, wherein the targeting lipid comprises a plurality of N-acetyl galactosamine moieties.

23. The formulation of claim 21, wherein said targeting lipid is present in the formulation in a molar amount of from about 0.001% to about 5%.

24. The formulation of claim 23, wherein said targeting lipid is present in the formulation in a molar amount of from about 0.005% to about 1.5%.

25. The formulation of claim 21, wherein said targeting lipid is the compound selected from the group consisting of formula 2, formula 3, formula 5, formula 6 or formula 7:

26. The lipid formulation of claim 6, comprising about 50% of cationic lipid of formula A, about 10% of the neutral lipid, about 38.5% of the sterol, and about 1.5% of the PEG or PEG-modified lipid.

27. A method of delivering a therapeutic agent to a target gene comprising administering to a subject the lipid formulation of claim 11.

28. The method of claim 27, wherein the therapeutic agent is an RNA-based construct.

29. The method of claim 28, wherein the RNA-based construct is a dsRNA.

30. The method of claim 27, 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, topoisomerase I gene, topoisomerase II alpha gene, mutations in the p73 gene, mutations in the p21(WAF1/CIP1) gene, mutations in the p27(KIP1) gene, mutations in the PPM1D gene, mutations in the RAS gene, mutations in the caveolin I gene, mutations in the MIB I gene, mutations in the MTAI gene, mutations in the M68 gene, mutations in tumor suppressor genes, and mutations in the p53 tumor suppressor gene.

31. The method of claim 27, wherein the target gene is Factor VII.

32. The method of claim 27, further comprising comparing expression of the target gene with a preselected reference value.

33. The method of claim 27, wherein the therapeutic agent is an antisense, siRNA, ribozyme or microRNA.

34. The lipid formulation of claim 6 comprising about 57.1% of cationic lipid of formula A, about 7.1% of the neutral lipid, about 34.4% of the sterol, and about 1.4% of the PEG or PEG-modified lipid.

35. The formulation of claim 24, wherein said targeting lipid is present in the formulation in a molar amount of 0.3%.

36. The lipid formulation of claim 1, wherein the concentration of the cationic lipid of Formula A is between 45 and 55%.

37. The lipid formulation of claim 36, wherein the concentration of cationic lipid of Formula A is 50%.