Trialkyl cationic lipids and methods of use thereof
The present invention provides compositions and methods for the delivery of therapeutic agents to cells. In particular, these include novel, trialkyl, cationic lipids and nucleic acid-lipid particles that provide efficient encapsulation of nucleic acids and efficient delivery of the encapsulated nucleic acid to cells in vivo. The compositions of the present invention are highly potent, thereby allowing effective knock-down of a specific target protein at relatively low doses.
1. A lipid having a structural Formula (I):
X-A-Y—Z; (I)
or salts thereof, wherein:
X is alkylamino;
A is C 1 to C 6 optionally substituted alkyl, wherein said C 1 to C 6 optionally substituted alkyl can be saturated or unsaturated, and wherein A may or may not be present;
Y is selected from the group consisting of ketal, ester, optionally substituted carbamate, ether, and optionally substituted amide; and
Z has the formula:
wherein, R 1 , R 2 , and R 3 are each independently selected from the group consisting of C 8 to C 11 alkyl, wherein each of R 1 , R 2 , and R 3 can independently be saturated or unsaturated, and wherein each of R 1 , R 2 , and R 3 is optionally substituted.
2. The lipid of claim 1 , wherein each of the R 1 , R 2 , and R 3 alkyl chains has a length of from C 9 to C 10 .
3. The lipid of claim 1 , wherein at least one of the R 1 , R 2 , and R 3 alkyl chains comprises a cycloalkyl moiety and/or a double bond.
4. The lipid of claim 1 , wherein X is selected from the group consisting of dimethylamino, diethylamino and ethylmethylamino.
5. A lipid selected from the group consisting of:
or a salt thereof.
6. A lipid particle comprising a lipid of claim 1 .
7. The lipid particle of claim 6 , wherein the particle further comprises a non-cationic lipid.
8. The lipid particle of claim 7 , wherein the non-cationic lipid is selected from the group consisting of a phospholipid, cholesterol or a derivative thereof, or a mixture of a phospholipid and cholesterol or a derivative thereof.
9. The lipid particle of claim 8 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), or a mixture thereof.
10. The lipid particle of claim 6 , wherein the particle further comprises a conjugated lipid that inhibits aggregation of particles.
11. The lipid particle of claim 10 , wherein the conjugated lipid that inhibits aggregation of particles comprises a polyethyleneglycol (PEG)-lipid conjugate.
12. The lipid particle of claim 6 wherein the particle further comprises a therapeutic agent.
13. The lipid particle of claim 12 , wherein the therapeutic agent is an interfering RNA selected from the group consisting of a small interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a microRNA (miRNA), a Dicer-substrate dsRNA, a small hairpin RNA (shRNA), and mixtures thereof.
14. The lipid particle of claim 12 , wherein the particle has a lipid:therapeutic agent mass ratio of from about 5:1 to about 15:1.
15. A pharmaceutical composition comprising a lipid particle of claim 6 a pharmaceutically acceptable carrier.
16. A method for introducing a therapeutic agent into a cell, the method comprising:
contacting the cell with a lipid particle of claim 12 .
17. A method for the in vivo delivery of a therapeutic agent, the method comprising:
administering to a mammal a lipid particle of claim 12 .
18. A method for treating a disease or disorder in a mammal in need thereof, the method comprising:
administering to the mammal a therapeutically effective amount of a lipid particle of claim 12 .
19. The method of claim 18 , wherein the disease or disorder is selected from the group consisting of a viral infection, a liver disease or disorder, and cancer.
20. The lipid of claim 1 , wherein when A, R 1 , R 2 , and R 3 are each independently optionally substituted, at least one hydrogen atom is replaced with a substituent selected from oxo, halogen, heterocycle, —CN, —OR x , —NR x R y , —NR x C(═O)R y , —NR x SO 2 R y , —C(═O)R x , —C(═O)OR x , —C(═O)NR x R y , —SO n R x , and —SO n NR x R y , wherein n is 0, 1, or 2, R x and R y are the same or different and are independently hydrogen, alkyl, or heterocycle, and each of the alkyl and heterocycle substituents may be further substituted with one or more of oxo, halogen, —OH, —CN, alkyl, —OR x , heterocycle, —NR x R y , —NR x C(═O)R y , —NR x SO 2 R y , —C(═O)R x , —C(═O)OR x , —C(═O)NR x R y , —SO n R x , and —SO n NR x R y ; and wherein when Y is independently optionally substituted, Y is independently optionally substituted with a saturated or unsaturated alkyl group.
21. The lipid of claim 1 , wherein when A, Y, R 1 , R 2 , and R 3 are not substituted.