Non-liposomal systems for nucleic acid delivery
View Patent ↗The present invention provides novel, stable lipid particles having a non-lamellar structure and comprising one or more active agents or therapeutic agents, methods of making such lipid particles, and methods of delivering and/or administering such lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) that have a non-lamellar structure and that comprise a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP.
1. A composition comprising:
a plurality of nucleic acid-lipid particles, wherein each particle in the plurality of particles comprises:
(a) a nucleic acid;
(b) a cationic lipid comprising from about 50 mol % to about 85 mol % of the total lipid present in the particle;
(c) a non-cationic lipid comprising from about 13 mol % to about 49.5 mol % of the total lipid present in the particle; and
(d) a conjugated lipid that inhibits aggregation of particles comprising from about 0.5 mol % to about 10 mol % of the total lipid present in the particle,
wherein at least about 95% of the particles in the plurality of particles have a non-lamellar morphology.
2. The composition of claim 1 , wherein the nucleic acid is an interfering RNA selected from the group consisting of siRNA, aiRNA, miRNA, Dicer-substrate dsRNA, shRNA, ssRNAi oligonucleotides, and combinations thereof.
3. The composition of claim 1 , wherein the non-cationic lipid is a mixture of a phospholipid and cholesterol or a cholesterol derivative.
4. The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-lipid conjugate.
5. The composition of claim 4 , wherein the PEG-lipid conjugate is member selected from the group consisting of: a PEG-diacylglycerol (PEG-DAG) conjugate, a PEG dialkyloxypropyl (PEG-DAA) conjugate, a PEG-phospholipid conjugate, a PEG-ceramide (PEG-Cer) conjugate, and a mixture thereof.
6. The composition of claim 1 , wherein the nucleic acid is fully encapsulated in the particles.
7. The composition of claim 1 , wherein the non-lamellar morphology of the particles comprises an inverse hexagonal (H II ) or cubic phase structure.
8. The composition of claim 7 , wherein the particles having a non-lamellar morphology are electron-dense.
9. The composition of claim 1 , wherein the non-lamellar morphology of the particles is determined by cryogenic-temperature transmission electron microscopy (cryo-TEM).
10. The composition of claim 1 , wherein the non-lamellar morphology of the particles is determined by X-ray diffraction.
11. The composition of claim 1 , wherein the non-lamellar morphology of the particles is determined by Differential Scanning calorimetry (DSC), wherein no thermal transitions are seen at 5 to 75° C.
12. A pharmaceutical composition comprising a composition of claim 1 and a pharmaceutically acceptable carrier.
13. A method for introducing a therapeutic agent into a cell, the method comprising:
contacting the cell with a composition of claim 1 .
14. A method for the in vivo delivery of a therapeutic agent, the method comprising:
administering to a mammal a composition of claim 1 .
15. The composition of claim 1 , wherein the nucleic acid is mRNA.
16. The composition of claim 1 , wherein the cationic lipid comprises from about 50 mol % to about 65 mol % of the total lipid present in the particle.
17. The composition of claim 1 , wherein the cationic lipid comprises from about 50 mol % to about 60 mol % of the total lipid present in the particle.
18. The composition of claim 3 , wherein the phospholipid comprises from about 3 mol % to about 15 mol % of the total lipid present in the particle.
19. The composition of claim 3 , wherein the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle.
20. The composition of claim 3 , wherein the cholesterol or derivative thereof comprises from about 30 mol % to about 40 mol % of the total lipid present in the particle.
21. The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of the particles comprises from about 0.5 mol % to about 2 mol % of the total lipid present in the particle.
22. The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of the particles comprises from about 5 mol % to about 10 mol % of the total lipid present in the particle.
23. The composition of claim 5 , wherein the PEG-DAA conjugate is a member selected from the group consisting of a PEG-didecyloxypropyl (C 10 ) conjugate, a PEG-dilauryloxypropyl (C 12 ) conjugate, a PEG-dimyristyloxypropyl (C 14 ) conjugate, a PEG-dipalmityloxypropyl (C 16 ) conjugate, a PEG-distearyloxypropyl (C 18 ) conjugate, and a mixture thereof.
24. The composition of claim 1 , wherein greater than 95% of the particles have a non-lamellar morphology.