Polyethyleneglycol-modified lipid compounds and uses thereof

• US 8,936,942 B2
• Filed: 08/06/2010
• Issued: 01/20/2015
• Est. Priority Date: 09/15/2003
• Status: Active Grant

• Current Assignee: Arbutus Biopharma Corporation
• Original Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Inventors: Heyes, James, MacLachlan, Ian, Ambegia, Ellen Grace
• Primary Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US12/852,362
• Publication Number:
  US 20110091525A1
• Time in Patent Office: 1,628 Days
• Field of Search: 424/93.2, 424/450, 424/489, 435/325, 435/375, 435/455, 435/366, 435/458, 514/44.A, 514/44.R
• US Class Current: 435/458
• 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. A liposome, said liposome comprising a polyethyleneglycol-dialkyloxypropyl (PEG-DAA) conjugate of Formula I having the following structure:

2. The liposome in accordance with claim 1, further comprising a bioactive agent.

3. The liposome in accordance with claim 2, wherein said bioactive agent is a nucleic acid.

4. A method of delivering a bioactive agent to a cell, said method comprising contacting said cell with a liposome of claim 1, wherein said bioactive agent is encapsulated in said liposome.

5. A method of delivering a bioactive agent to a patient, said method comprising administering to said patient a liposome of claim 1, wherein said bioactive agent is encapsulated in said liposome.

36. The liposome in accordance with claim 1, wherein the non-ester containing linker is a member selected from the group consisting of an amido linker moiety, an amino linker moiety, a carbonyl linker moiety, a carbamate linker moiety, a urea linker moiety, an ether linker moiety, a disulphide linker moiety, a succinamidyl linker moiety and combinations thereof.

38. The liposome in accordance with claim 1, wherein the non-ester containing linker is a member selected from the group consisting of an amido linker moiety, an amino linker moiety, a carbonyl linker moiety, a urea linker moiety, an ether linker moiety, a disulphide linker moiety, a succinamidyl linker moiety and combinations thereof.

6. 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;

7. The nucleic acid-lipid particle in accordance with claim 6, 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-dimethy1-2,3-dioleyloxy)propylamine (DODMA), and a mixture thereof.

8. The nucleic acid-lipid particle in accordance with claim 6, 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.

9. The nucleic acid-lipid particle in accordance with claim 6, wherein said non-cationic lipid is an anionic lipid.

10. The nucleic acid-lipid particle in accordance with claim 6, wherein said non-cationic lipid is a neutral lipid.

11. The nucleic acid-lipid particle in accordance with claim 6, wherein said PEG-DAA conjugate is a member selected from the group consisting of PEG-dilauryloxypropyl (C 12), a PEG-dimyristyloxypropyl (C14), a PEG-dipalmityloxypropyl (C16), and a PEG-disteryloxypropyl (C18).

12. The nucleic acid-lipid particle in accordance with claim 6, wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present in said particle.

13. The nucleic acid-lipid particle in accordance with claim 6, wherein said cationic lipid comprises from about 5% to about 45% of the total lipid present in said particle.

14. The nucleic acid-lipid particle in accordance with claim 6, wherein said cationic lipid comprises from about 5% to about 15% of the total lipid present in said particle.

15. The nucleic acid-lipid particle in accordance with claim 6, wherein said cationic lipid comprises from about 40% to about 50% of the total lipid present in said particle.

16. The nucleic acid-lipid particle in accordance with claim 6, wherein said non-cationic lipid comprises from about 5% to about 90% of the total lipid present in said particle.

17. The nucleic acid-lipid particle in accordance with claim 6, wherein said non-cationic lipid comprises from about 20% to about 85% of the total lipid present in said particle.

18. The nucleic acid-lipid particle in accordance with claim 6, wherein said PEG-DAA conjugate comprises from 1% to about 20% of the total lipid present in said particle.

19. The nucleic acid-lipid particle in accordance with claim 6, wherein said PEG-DAA 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 6, wherein said PEG-DAA conjugate comprises from 4% to about 10% of the total lipid present in said particle.

21. The nucleic acid-lipid particle in accordance with claim 6, wherein said non-cationic lipid is DSPC.

22. The nucleic acid-lipid particle in accordance with claim 6, further comprising cholesterol.

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

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

25. The nucleic acid-lipid particle in accordance with claim 6, wherein the PEG-DAA conjugate is PEG-dimyristyloxypropyl (C14).

26. The nucleic acid-lipid particle in accordance with claim 6, wherein said nucleic acid is an antisense oligonucleotide.

27. The nucleic acid-lipid particle in accordance with claim 6, wherein said nucleic acid is a ribozyme.

28. The nucleic acid-lipid particle in accordance with claim 6, wherein said nucleic acid is a small interfering RNA (siRNA).

29. The nucleic acid-lipid particle in accordance with claim 6, wherein said therapeutic product of interest is a small interfering RNA (si RNA).

30. The nucleic acid-lipid particle in accordance with claim 6, 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.

31. The nucleic acid-lipid particle in accordance with claim 6, 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.

32. The nucleic acid-lipid particle in accordance with claim 6, wherein the nucleic acid is fully encapsulated in said nucleic acid-lipid particle.

33. A pharmaceutical composition comprising a nucleic acid-lipid particle in accordance with claim 6 and a pharmaceutically acceptable carrier.

34. A pharmaceutical composition in accordance with claim 33, wherein the PEG-DAA conjugate is PEG-dimyristyloxypropyl (C14).

37. The nucleic acid-lipid particle in accordance with claim 6, wherein the non-ester containing linker is a member selected from the group consisting of an amido linker moiety, an amino linker moiety, a carbonyl linker moiety, a carbamate linker moiety, a urea linker moiety, an ether linker moiety, a disulphide linker moiety, a succinamidyl linker moiety and combinations thereof.

39. The nucleic acid-lipid particle in accordance with claim 6, wherein the non-ester containing linker is a member selected from the group consisting of an amido linker moiety, an amino linker moiety, a carbonyl linker moiety, a urea linker moiety, an ether linker moiety, a disulphide linker moiety, a succinamidyl linker moiety and combinations thereof.

35. 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.