POLYETHYLENEGLYCOL-MODIFIED LIPID COMPOUNDS AND USES THEREOF

• US 20110091525A1
• Filed: 08/06/2010
• Published: 04/21/2011
• Est. Priority Date: 09/15/2003
• Status: Active Grant

• Current Assignee: Arbutus Biopharma Corporation
• Original Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Inventors: Ambegia, Ellen Grace, Heyes, James, MacLachlan, Ian
• Granted Patent: US 8,936,942 B2
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/6911 the form being a liposome
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07C 233/18 having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
• CPC Class Codes: C07C 235/08 having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
• CPC Class Codes: C07C 271/16 to carbon atoms of hydrocarbon radicals substituted by singly-bound oxygen atoms
• CPC Class Codes: C08L 2203/02 for biomedical use

Abstract

The present invention provides compositions comprising polytheylyene-dialkyloxypropyl conjugates (PEG-DAA), liposomes, SNALP, and SPLP comprising such compositions, and methods of using such compositions, liposomes, SNALP, and SPLP.

Claims

1-17. -17. (canceled)

18. A liposome, said liposome comprising a polyethyleneglycol-dialkyloxypropyl (PEG-DAA) conjugate of Formula I having the following structure:

19. The liposome in accordance with claim 18, further comprising a bioactive agent.

20. The liposome in accordance with claim 19, wherein said bioactive agent is a member selected from the group consisting of antineoplastic agents, antibiotics, immunomodulators, anti-inflammatory agents and agents acting on the central nervous system.

21. The liposome in accordance with claim 19, wherein said bioactive agent is a protein or peptide.

22. The liposome in accordance with claim 19, wherein said bioactive agent is a nucleic acid.

23. A method of delivering a bioactive agent to a cell, said method comprising contacting said cell with a liposome of claim 18, wherein said bioactive agent is encapsulated in said liposome.

24. A method of delivering a bioactive agent to a patient, said method comprising administering to said patient a liposome of claim 18, wherein said bioactive agent is encapsulated in said liposome.

25. A nucleic acid-lipid particle comprising:

a nucleic acid;

a cationic lipid;

a non-cationic lipid; and

a polyethyleneglycol-dialkyloxypropyl (PEG-DAA) conjugate of Formula I having the following structure;

26. The nucleic acid-lipid particle in accordance with claim 25, wherein said cationic lipid is a member selected from the group consisting of N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), and N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), and a mixture thereof.

27. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid is a member selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), palmitoyloleyolphosphatidylglycerol (POPG), cholesterol, and a mixture thereof.

28. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid is an anionic lipid.

29. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid is a neutral lipid.

30. The nucleic acid-lipid particle in accordance with claim 25, wherein said PEG-DAA conjugate is a member selected from the group consisting of PEG-dilauryloxypropyl (C12), a PEG-dimyristyloxypropyl (C14), a PEG-dipalmityloxypropyl (C16), and a PEG-disteryloxypropyl (C18).

31. The nucleic acid-lipid particle in accordance with claim 25, wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present in said particle.

32. The nucleic acid-lipid particle in accordance with claim 25, wherein said cationic lipid comprises from about 5% to about 45% of the total lipid present in said particle.

33. The nucleic acid-lipid particle in accordance with claim 25, wherein said cationic lipid comprises from about 5% to about 15% of the total lipid present in said particle.

34. The nucleic acid-lipid particle in accordance with claim 25, wherein said cationic lipid comprises from about 40% to about 50% of the total lipid present in said particle.

35. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid comprises from about 5% to about 90% of the total lipid present in said particle.

36. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid comprises from about 20% to about 85% of the total lipid present in said particle.

37. The nucleic acid-lipid particle in accordance with claim 25, wherein said PEG-DAA conjugate comprises from 1% to about 20% of the total lipid present in said particle.

38. The nucleic acid-lipid particle in accordance with claim 25, wherein said PEG-DAA conjugate comprises from 2% to about 15% of the total lipid present in said particle.

39. The nucleic acid-lipid particle in accordance with claim 25, wherein said PEG-DAA conjugate comprises from 4% to about 10% of the total lipid present in said particle.

40. The nucleic acid-lipid particle in accordance with claim 25, wherein said non-cationic lipid is DSPC.

41. The nucleic acid-lipid particle in accordance with claim 25, further comprising cholesterol.

42. The nucleic acid-lipid particle in accordance with claim 41, wherein the cholesterol comprises from about 10% to about 60% of the total lipid present in said particle.

43. The nucleic acid-lipid particle in accordance with claim 41, wherein the cholesterol comprises from about 20% to about 45% of the total lipid present in said particle.

44. The nucleic acid-lipid particle in accordance with claim 25, wherein the PEG-DAA conjugate is PEG-dilauryloxypropyl (C12).

45. The nucleic acid-lipid particle in accordance with claim 25, wherein the PEG-DAA conjugate is PEG-dimyristyloxypropyl (C14).

46. The nucleic acid-lipid particle in accordance with claim 25, wherein the PEG-DAA conjugate is PEG-dipalmityloxypropyl (C16).

47. The nucleic acid-lipid particle in accordance with claim 25, wherein the PEG-DAA conjugate is PEG-disteryloxypropyl (C18).

48. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid is DNA.

49. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid is a plasmid.

50. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid is an antisense oligonucleotide.

51. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid is a ribozyme.

52. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid is a small interfering RNA (siRNA).

53. The nucleic acid-lipid particle in accordance with claim 25, wherein said nucleic acid encodes a therapeutic product of interest.

54. The nucleic acid-lipid particle in accordance with claim 53, wherein said therapeutic product of interest is a peptide or protein.

55. The nucleic acid-lipid particle in accordance with claim 53, wherein said therapeutic product of interest is a small interfering RNA (siRNA).

56. The nucleic acid-lipid particle in accordance with claim 25, wherein the nucleic acid in said nucleic acid-lipid particle is not substantially degraded after exposure of said particle to a nuclease at 37°

C. for 20 minutes.

57. The nucleic acid-lipid particle in accordance with claim 25, wherein the nucleic acid in said nucleic acid-lipid particle is not substantially degraded after incubation of said particle in serum at 37°

C. for 30 minutes.

58. The nucleic acid-lipid particle in accordance with claim 25, wherein the nucleic acid is fully encapsulated in said nucleic acid-lipid particle.

59. A pharmaceutical composition comprising a nucleic acid-lipid particle in accordance with claim 25 and a pharmaceutically acceptable carrier.60. (Original) A pharmaceutical composition in accordance with claim 59, wherein the PEG-DAA conjugate is PEG-dimyristyloxypropyl (C14).

61. A pharmaceutical composition in accordance with claim 59, wherein the PEG-DAA conjugate is PEG-disteryloxypropyl (C18).

62. A method of introducing a nucleic acid into a cell, said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid, a non-cationic lipid, a PEG-DAA conjugate, and a nucleic acid. wherein the wavy line indicates the point of attachment to the rest of the compound.