Polyethyleneglycol-modified lipid compounds and uses thereof
View Patent ↗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.
1. A liposome, said liposome comprising a polyethyleneglycol-dialkyloxypropyl (PEG-DAA) conjugate of Formula I having the following structure:
wherein:
R l and R 2 are independently selected and are alkyl groups having from about 10 to about 20 carbon atoms;
PEG is a polyethyleneglycol, wherein the terminal hydroxyl group is substituted with a methyl group; and
L is a non-ester containing linker moiety.
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.
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:
wherein:
R l and R 2 are independently selected and are alkyl groups having from about 10 to about 20 carbon atoms;
PEG is a polyethyleneglycol, wherein the terminal hydroxyl group is substituted with a methyl group; and
L is a non-ester containing linker moiety.
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).
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.
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.
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.
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.
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.