Compositions for the delivery of therapeutic agents and uses thereof

• US 20060051405A1
• Filed: 07/19/2005
• Published: 03/09/2006
• Est. Priority Date: 07/19/2004
• Status: Abandoned Application

• Current Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Original Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Inventors: MacLachlan, Ian, Jeffs, Lloyd
• Application Number:
  US11/185,447
• Publication Number:
  US 20060051405A1
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• 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: 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 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

The present invention provides drug delivery vehicles comprising polytheylyene-lipid conjugates (PEG-lipid), wherein the circulation lifetime and biodistribution of the drug delivery vehicles are regulated by the PEG-lipid. More particularly, the present invention provides liposomes, SNALP and SPLP comprising such PEG-lipid conjugates, and methods of using such compositions to selectively target a tumor site or other tissue of interest (e.g., liver, lung, spleen, etc.).

Claims

1. A method of introducing a nucleic acid into a tumor cell, said method comprising contacting said tumor cell with a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 12 to 20 carbon atoms.

2. The method in accordance with claim 1, wherein said cationic lipid is a member selected from the group consisting of:

1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 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.

3. The method in accordance with claim 1, wherein said noncationic lipid is a member selected from the group consisting of:

(DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl- phosphatidylethanolamine (POPE) and dioleoyl- phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), and 1,2-dielaidoyl-sn-glycero-3-phophoethanolamine (transDOPE).

4. The method in accordance with claim 1, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 16 to 20 carbon atoms.

5. The method in accordance with claim 1, wherein said noncationic lipid is an anionic lipid.

6. The method in accordance with claim 1, wherein said noncationic lipid is a neutral lipid.

7. The method in accordance with claim 1, wherein said PEG-lipid is a member selected from the group consisting of a PEG-diacylglycerol (DAG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide, and combinations thereof.

8. The method in accordance with claim 1, wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) selected from the group consisting of:

a PEG-dipalmityloxypropyl (C₁₆);

a PEG-distearyloxypropyl (C₁₈); and

a PEG-diicosyloxypropyl (C₂₀).

9. The method in accordance with claim 1, wherein said PEG-lipid is PEG-dialkyloxypropyl (DAA) having the following structure:

wherein;

R¹ and R² are independently selected and are alkyl groups having from about 16 to about 20 carbon atoms;

PEG is a polyethyleneglycol; and

L is a non-ester containing linker moiety.

10. The method in accordance with claim 1, wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of a PEG-dipalmitoylglycerol (C₁₆), a PEG-disterylglycerol (C₁₈) and a PEG-diicosylglycerol (C20).

11. The method in accordance with claim 1, wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of PEG-ceramide (C₁₆), a PEG-ceramide (C₁₈) and PEG-ceramide (C₂₀).

12. The method in accordance with claim 1, wherein said cationic lipid comprises from about 5% to about 45% of the total lipid present in said particle.

13. The method in accordance with claim 1, wherein said cationic lipid comprises from about 5% to about 15% of the total lipid present in said particle.

14. The method in accordance with claim 1, wherein said cationic lipid comprises from about 30% to about 50% of the total lipid present in said particle.

15. The method in accordance with claim 1, wherein said cationic lipid comprises about 40% of the total lipid present in said particle.

16. The method in accordance with claim 1, wherein said noncationic lipid comprises from about 5% to about 90% of the total lipid present in said particle.

17. The method in accordance with claim 1, wherein said noncationic lipid comprises from about 20% to about 85% of the total lipid present in said particle.

18. The method in accordance with claim 1, wherein said PEG-lipid conjugate comprises from 1% to about 20% of the total lipid present in said particle.

19. The method in accordance with claim 1, wherein said PEG-lipid conjugate comprises from 2% to about 15% of the total lipid present in said particle.

20. The nucleic acid-lipid particle in accordance with claim 1, wherein said PEG-lipid conjugate comprises about 2% of the total lipid present in said particle.

21. The method in accordance with claim 1, wherein said noncationic lipid is DSPC.

22. The method in accordance with claim 1, wherein said nucleic acid-lipid particle further comprises cholesterol.

23. The method in accordance with claim 22, wherein the cholesterol comprises from about 0% to about 10% of the total lipid present in said particle.

24. The method in accordance with claim 22, wherein the cholesterol comprises from about 10% to about 60% of the total lipid present in said particle.

25. The method in accordance with claim 22, wherein the cholesterol comprises from about 20% to about 45% of the total lipid present in said particle.

26. The method in accordance with claim 1, wherein said nucleic acid is DNA.

27. The method in accordance with claim 1, wherein said nucleic acid is a plasmid.

28. The method in accordance with claim 1, wherein said nucleic acid is an antisense oligonucleotide.

29. The method in accordance with claim 1, wherein said nucleic acid is a ribozyme.

30. The method in accordance with claim 1, wherein said nucleic acid is a small interfering RNA (siRNA).

31. The method in accordance with claim 1, wherein said nucleic acid encodes a therapeutic product of interest.

32. The method in accordance with claim 31, wherein said therapeutic product of interest is a peptide or protein.

33. The method in accordance with claim 31, wherein said therapeutic product of interest is a small interfering RNA (siRNA).

34. The method in accordance with claim 1, 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.

35. The method in accordance with claim 1, 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.

36. The method in accordance with claim 1, wherein the nucleic acid is fully encapsulated in said nucleic acid-lipid particle.

37. A method of introducing a nucleic acid to the lung of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 16 to 20 carbon atoms.

38. The method in accordance with claim 37, wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dipalmityloxypropyl (C₁₆), PEG-distearyloxypropyl (C₁₈), and a PEG-diicosyloxypropyl (C₂₀).

39. The method in accordance with claim 37, wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of:

a PEG-dipalmitoylglycerol (C₁₆), and a PEG-disterylglycerol (C₁₈).

40. The method in accordance with claim 37, wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of:

PEG-ceramide (C₁₆), and a PEG-ceramide (C₁₈) and PEG-ceramide (C₂₀).

41. A method of introducing a nucleic acid to the liver of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 8 to 14 carbon atoms.

42. The method in accordance with claim 41, wherein said PEG-lipid is as PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dilauryloxypropyl (C₁₂) and a PEG-dimyristyloxypropyl (C₁₄).

43. The method in accordance with claim 41, wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of:

PEG-dilaurylglycerol (C₁₂) and a PEG-dimyristylglycerol (C₁₄).

44. The method in accordance with claim 41, wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of:

PEG-ceramide (C₁₂) and a PEG-ceramide (C₁₄).

45. A method of introducing a nucleic acid to the spleen of a mammal, said method comprising administering to said mammal a nucleic acid-lipid particle comprising a cationic lipid, a noncationic lipid, a PEG-lipid conjugate, and a nucleic acid, wherein the alkyl or acyl chains of the lipid portion of said PEG-lipid conjugate comprise from 8 to 14 carbon atoms.

46. The method in accordance with claim 45, wherein said PEG-lipid is a PEG dialkyloxypropyl (DAA) slected from the group consisting of a PEG-dilauryloxypropyl (C₁₂) and a PEG-dimyristyloxypropyl (C₁₄).

47. The method in accordance with claim 45, wherein said PEG-lipid is PEG-diacylglycerol (DAG) selected from the group consisting of:

PEG-dilaurylglycerol (C₁₂) and a PEG-dimyristylglycerol (C₁₄).

48. The method in accordance with claim 45, wherein said PEG-lipid is PEG-ceramide (Cer) selected from the group consisting of selected from the group consisting of PEG-ceramide (C₁₂) and a PEG-ceramide (C₁₄).