Lipid formulations for nucleic acid delivery
View Patent ↗The present invention provides novel, stable lipid particles comprising one or more active agents or therapeutic agents, methods of making the lipid particles, and methods of delivering and/or administering the lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) comprising 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 nucleic acid-lipid particle comprising:
(a) a nucleic acid;
(b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle;
(c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof, wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle; and
(d) a conjugated lipid that inhibits aggregation of particles comprising from 0.5 mol % to 2 mol % of the total lipid present in the particle.
2. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises a small interfering RNA (siRNA).
3. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises from about 15 to about 60 nucleotides.
4. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises at least one modified nucleotide.
5. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises at least one 2′-O-methyl (2′OMe) nucleotide.
6. The nucleic acid-lipid particle of claim 2 , wherein said siRNA is about 19 to about 25 base pairs in length.
7. The nucleic acid-lipid particle of claim 2 , wherein said siRNA comprises 3′ overhangs.
8. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present in the particle.
9. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), or a mixture thereof.
10. The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises a polyethyleneglycol (PEG)-lipid conjugate.
11. The nucleic acid-lipid particle of claim 10 , wherein the PEG-lipid conjugate comprises a PEG-diacylglycerol (PEG-DAG) conjugate, a PEG-dialkyloxypropyl (PEG-DAA) conjugate, or a mixture thereof.
12. The nucleic acid-lipid particle of claim 11 , wherein the PEG-DAA conjugate comprises a PEG-dimyristyloxypropyl (PEG-DMA) conjugate, a PEG-distearyloxypropyl (PEG-DSA) conjugate, or a mixture thereof.
13. The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight of about 2,000 daltons.
14. The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57.1 mol % cationic lipid, about 7.1 mol % phospholipid, about 34.3 mol % cholesterol or a derivative thereof, and about 1.4 mol % PEG-lipid conjugate.
15. The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle.
16. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C. for 30 minutes.
17. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid is fully encapsulated in the nucleic acid-lipid particle.
18. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid:nucleic acid mass ratio of from about 5 to about 15.
19. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter of from about 40 nm to about 150 nm.
20. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present in the particle.
21. The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present in the particle.
22. A pharmaceutical composition comprising a nucleic acid-lipid particle of claim 1 and a pharmaceutically acceptable carrier.