Non-liposomal systems for nucleic acid delivery
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 a mixture of: (i) a phospholipid of from about 3 mol % to about 15 mol % of the total lipid present in the particle; and (ii) cholesterol or a derivative thereof of from about 30 mol % to about 40 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 are electron-dense; and
wherein the size of the particles has a polydispersity value of about 0.03.
2. The composition of claim 1 , wherein the nucleic acid is mRNA.
3. The composition of claim 1 , wherein the nucleic acid is an interfering RNA.
4. The composition of claim 1 , wherein the nucleic acid is fully encapsulated in the particles.
5. 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.
6. 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.
7. The composition of claim 1 , wherein the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle.
8. The composition of claim 1 , wherein the phospholipid comprises from about 4 mol % to about 10 mol % of the total lipid present in the particle.
9. The composition of claim 1 , wherein the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle.
10. 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.
11. The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of the particles comprises from about 4 mol % to about 10 mol % of the total lipid present in the particle.
12. 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 and the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle.
13. 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 and the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle.
14. 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 and 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.
15. 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, the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle, and the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle.
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, the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle, and 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.
17. 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, the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle, and 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.
18. 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, the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle, the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle, and 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.
19. The composition of claim 1 , wherein the phospholipid comprises from about 4 mol % to about 12 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from about 30 mol % to about 35 mol % of the total lipid present in the particle.
20. The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-lipid conjugate.
21. The composition of claim 20 , wherein the PEG-lipid conjugate is 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.
22. The composition of claim 21 , 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.
23. The composition of claim 20 , wherein the PEG has an average molecular weight of about 2,000 daltons.
24. The composition of claim 23 , wherein the PEG comprises a terminal methoxy group.
25. The composition of claim 23 , wherein the PEG is linked to the lipid via an ether moiety.
26. The composition of claim 1 , wherein the electron-dense particles comprise an inverse hexagonal (H II ) or cubic phase structure.
27. The composition of claim 1 , wherein greater than 95% of the particles are electron-dense.
28. The composition of claim 1 , wherein at least 96% of the particles are electron-dense.
29. The composition of claim 1 , wherein at least 97% of the particles are electron-dense.
30. The composition of claim 1 , wherein at least 98% of the particles are electron-dense.
31. The composition of claim 1 , wherein at least 99% of the particles are electron-dense.
32. A pharmaceutical composition comprising a composition of claim 1 and a pharmaceutically acceptable carrier.