LIPID FORMULATION

• US 20120183602A1
• Filed: 01/25/2012
• Published: 07/19/2012
• Est. Priority Date: 06/10/2009
• Status: Active Grant

• Current Assignee: Arbutus Biopharma Corporation
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Ansell, Steven, Akinc, Akin, Dorkin, Joseph Robert, Qin, Xiaojun, Cantley, William, Manoharan, Muthiah, Rajeev, Kallanthottathil G., Narayanannair, Jayaprakash K., Jayaraman, Muthusamy, CHEN, Jianxin
• Granted Patent: US 8,802,644 B2
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/22 of acyclic acids, e.g. pravastatin
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/712 Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: A61K 47/14 Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/551 one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
• CPC Class Codes: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• 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: A61K 9/1278 Post-loading, e.g. by ion or pH gradient
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07C 227/18 by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
• CPC Class Codes: C07C 227/40 Separation; Purification
• CPC Class Codes: C07C 229/12 to carbon atoms of acyclic carbon skeletons
• CPC Class Codes: C07C 29/58 by elimination of halogen, e.g. by hydrogenolysis, splitting-off C07C29/124 takes precedence
• CPC Class Codes: C07D 475/04 : with a nitrogen atom directly attached in position 2
• 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 15/1137 : against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• 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 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/341 : Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 : having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12N 2310/3515 : Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2310/3533 : Halogen
• CPC Class Codes: C12N 2320/32 : Special delivery means, e.g. tissue-specific
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention features a cationic lipid of formula I,

• • an improved lipid formulation comprising a cationic lipid of formula I and corresponding methods of use.

Also disclosed are targeting lipids, and specific lipid formulations comprising such targeting lipids.

Claims

1. (canceled)

2. A lipid formulation comprising:

(a) a compound having the formula

3. The lipid formulation of claim 2, comprising:

(a) 40-65% of the compound,

(b) 5-10% of the neutral lipid,

(c) 25-40% of the sterol, and

(d) 0.5-10% of the PEG-modified lipid.

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

5. The lipid formulation of claim 2, wherein the sterol is cholesterol.

6. The lipid formulation of claim 2, wherein the PEG lipid is PEG-C₁₄ to PEG-C₂₂, PEG-Cer₁₄ to PEG-C₂₀, or PEG-DSPE.

8. The lipid formulation of claim 2, comprising:

(a) about 57.5% of the compound,

(b) about 7.5% of the neutral lipid,

(c) about 31.5% of the sterol, and

(d) about 3.5% of the PEG-modified lipid.

11. The lipid formulation of claim 2, further comprising a therapeutic agent, which comprises a nucleic acid.

12. The lipid formulation of claim 11, wherein the nucleic acid is an siRNA.

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

nucleic acid is about 3 to about 15.

14. The lipid formulation of claim 13, wherein the molar ratio of lipid:

nucleic acid about 5 to about 13.

22. The lipid formulation of claim 2, comprising:

(a) about 50% of the compound,

(b) about 10% of the neutral lipid,

(c) about 38.5% of the sterol, and

(d) about 1.5% of the PEG-modified lipid.

23. The lipid formulation of claim 2, comprising:

(a) about 50% of the compound,

(b) about 10% of the neutral lipid,

(c) about 35% of the sterol, and

(d) about 5% of the PEG-modified lipid.

24. The lipid formulation of claim 2, comprising:

(a) about 57.2% of the compound,

(a) about 7.1% of the neutral lipid,

(a) about 34.3% of the sterol, and

(a) 1 about 1.4% of the PEG-modified lipid.

25. A method of delivering an siRNA therapeutic agent to a cell comprising administering to a subject the lipid formulation of claim 12.

26. The method of claim 25, wherein the siRNA therapeutic agent is a dsRNA.

30. A method of modulating the expression of a target gene in a cell, the method comprising providing to a cell the lipid formulation of claim 12.

31. The lipid formulation of claim 2, comprising:

(a) 20-60% of the compound cationic lipid of formula I,

(b) 5-25% of the neutral lipid,

(c) 25-55% of the sterol, and

(d) 0.5-15% of the PEG-modified lipid.

32. The lipid formulation of claim 2, comprising:

(a) 20-75% of the compound cationic lipid of formula I,

(b) 5-15% of the neutral lipid,

(c) 5-50% of the sterol, and

(d) 0.5-20% of the PEG-modified lipid.

33. The lipid formulation of claim 2, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

34. The lipid formulation of claim 8, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

35. The lipid formulation of claim 22, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

36. The lipid formulation of claim 23, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

37. The lipid formulation of claim 24, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

38. The lipid formulation of claim 25, wherein the neutral lipid is DSPC, the sterol is cholesterol, the PEG-modified lipid is PEG-DMG.

39. The lipid formulation of claim 33, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

40. The lipid formulation of claim 34, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

41. The lipid formulation of claim 35, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

42. The lipid formulation of claim 36, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

43. The lipid formulation of claim 37, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

44. The lipid formulation of claim 38, wherein the PEG moiety of the PEG-DMG has an average molecular weight of 2,000 Da.

45. The lipid formulation of claim 11, wherein the formulation is prepared by a method comprising (i) extruding a liposome formed from components (a)-(d), and (ii) addition of a nucleic acid therapeutic agent to the liposome.

46. The lipid formulation of claim 11, wherein the formulation is prepared by a method comprising mixing components (a)-(d) and a nucleic acid therapeutic agent in parallel into a mixing chamber.

7. (canceled)

9-10. -10. (canceled)

15-21. -21. (canceled)

27-29. -29. (canceled)

47. A nucleic acid-lipid particle comprising:

(a) a lipid particle comprising;

(i) a compound having the formula

48. The nucleic acid-lipid particle of claim 47, wherein the nucleic acid-lipid particle has a size ranging from about 70 to about 200 nm.

49. The nucleic acid-lipid particle of claim 48, wherein the nucleic acid-lipid particle has a size ranging from about 90 to about 130 nm.