Soluble steroidal peptides for nucleic acid delivery
Amphiphilic lipopeptide compositions for gene delivery are disclosed. An illustrative amphiphilic lipopeptide composition includes a human protamine 2 peptide conjugated to a hydrophobic moiety. Illustrative hydrophobic moieties include sterols, bile acids, and fatty acids. The amphiphilic lipopeptide composition is mixed with a nucleic acid such that the nucleic acid binds to the peptide portion of the lipopeptide. This mixture is placed in contact with mammalian cells to effect transfection of the cells with the nucleic acid. A method of making such amphiphilic lipopeptides is also described.
1. A composition comprising a PRM2 peptide conjugated to a hydrophobic moiety.
2. The composition of claim 1 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2.
3. The composition of claim 1 wherein said hydrophobic moiety comprises a sterol.
4. The composition of claim 3 wherein said sterol is a member selected from the group consisting of cholestanol, coprostanol, cholesterol, epicholesterol, ergosterol, and ergocalciferol.
5. The composition of claim 3 wherein said sterol comprises cholesterol.
6. The composition of claim 1 wherein said hydrophobic moiety comprises a bile acid.
7. The composition of claim 6 wherein said bile acid is a member selected from the group consisting of cholic acid, deoxycholic acid, chenodeoxycholic acid, lithocholic acid, ursocholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, lagodeoxycholic acid, glycocholic acid, taurocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, dehydrocholic acid, hyocholic acid, and hyodeoxycholic acid.
8. The composition of claim 3 wherein said bile acid comprises lithocholic acid.
9. The composition of claim 1 wherein said hydrophobic moiety comprises a fatty acid.
10. The composition of claim 9 wherein said fatty acid is a member selected from the group consisting of C4-C20 alkanoic acids.
11. The composition of claim 9 wherein said fatty acid is a member selected from the group consisting of butyric acid, valeric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, and stearic acid.
12. The composition of claim 1 wherein said PRM2 peptide is conjugated to said hydrophobic moiety through a peptide linkage.
13. The composition of claim 12 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises cholesterol.
14. The composition of claim 12 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises lithocholic acid.
15. The composition of claim 1 wherein said hydrophobic moiety is conjugated to said PRM2 peptide through a non-terminal amino acid residue.
16. The composition comprising a mixture of a nucleic acid and a conjugate comprising a PRM2 peptide and a hydrophobic moiety.
17. The composition of claim 16 wherein said nucleic acid binds to said PRM2 peptide.
18. The composition of claim 16 wherein said nucleic acid comprises a plasmid.
19. The composition of claim 16 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2.
20. The composition of claim 16 wherein said hydrophobic moiety comprises a sterol.
21. The composition of claim 20 wherein said sterol is a member selected from the group consisting of cholestanol, coprostanol, cholesterol, epicholesterol, ergosterol, and ergocalciferol.
22. The composition of claim 20 wherein said sterol comprises cholesterol.
23. The composition of claim 16 wherein said hydrophobic moiety comprises a bile acid.
24. The composition of claim 23 wherein said bile acid is a member selected from the group consisting of cholic acid, deoxycholic acid, chenodeoxycholic acid, lithocholic acid, ursocholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, lagodeoxycholic acid, glycocholic acid, taurocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, dehydrocholic acid, hyocholic acid, and hyodeoxycholic acid.
25. The composition of claim 20 wherein said bile acid comprises lithocholic acid.
26. The composition of claim 16 wherein said hydrophobic moiety comprises a fatty acid.
27. The composition of claim 26 wherein said fatty acid is a member selected from the group consisting of C4-C20 alkanoic acids.
28. The composition of claim 26 wherein said fatty acid is a member selected from the group consisting of butyric acid, valeric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, and stearic acid.
29. The composition of claim 16 wherein said PRM2 peptide is conjugated to said hydrophobic moiety through a peptide linkage.
30. The composition of claim 29 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises cholesterol.
31. The composition of claim 29 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises lithocholic acid.
32. The composition of claim 16 wherein said hydrophobic moiety is conjugated to said PRM2 peptide through a non-terminal amino acid residue.
33. The composition of claim 16 wherein said nucleic acid encodes interleukin-12.
34. The composition of claim 33 wherein said nucleic acid comprises p2CMVmIL-12.
35. A method for transfecting a mammalian cell comprising contacting said cell with a composition comprising a mixture of a nucleic acid and a conjugate comprising a PRM2 peptide and a hydrophobic moiety, and then incubating said cell under conditions suitable for growth thereof.
36. The method of claim 35 wherein said mammalian cell is a human cell.
37. The method of claim 35 wherein said nucleic acid binds to said PRM2 peptide.
38. The method of claim 35 wherein said nucleic acid comprises a plasmid.
39. The method of claim 35 of claim 1 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2.
40. The method of claim 35 wherein said hydrophobic moiety comprises a sterol.
41. The method of claim 40 wherein said sterol is a member selected from the group consisting of cholestanol, coprostanol, cholesterol, epicholesterol, ergosterol, and ergocalciferol.
42. The method of claim 40 wherein said sterol comprises cholesterol.
43. The method of claim 35 wherein said hydrophobic moiety comprises a bile acid.
44. The method of claim 43 wherein said bile acid is a member selected from the group consisting of cholic acid, deoxycholic acid, chenodeoxycholic acid, lithocholic acid, ursocholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, lagodeoxycholic acid, glycocholic acid, taurocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, dehydrocholic acid, hyocholic acid, and hyodeoxycholic acid.
45. The method of claim 43 wherein said bile acid comprises lithocholic acid.
46. The method of claim 35 wherein said hydrophobic moiety comprises a fatty acid.
47. The method of claim 46 wherein said fatty acid is a member selected from the group consisting of C4-C20 alkanoic acids.
48. The method of claim 46 wherein said fatty acid is a member selected from the group consisting of butyric acid, valeric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, and stearic acid.
49. The method of claim 35 wherein said PRM2 peptide is conjugated to said hydrophobic moiety through a peptide linkage.
50. The method of claim 49 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises cholesterol.
51. The method of claim 49 wherein said PRM2 peptide comprises a peptide identified as SEQ ID NO:2, and said hydrophobic moiety comprises lithocholic acid.
52. The method of claim 35 wherein said hydrophobic moiety is conjugated to said PRM2 peptide through a non-terminal amino acid residue.
53. A method for making an amphiphilic lipopeptide comprises conjugating a hydrophobic moiety to a non-terminal amino acid residue of a PRM2 peptide.