Lipid formulations for delivery of therapeutic agents
The present invention provides novel, serum-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 serum-stable nucleic acid-lipid particles (SNALP) comprising a nucleic acid (e.g., one or more interfering RNA molecules), methods of making the SNALP, and methods of delivering and/or administering the SNALP (e.g., for the treatment of cancer). In particular embodiments, the present invention provides tumor-directed lipid particles that preferentially target solid tumors. The tumor-directed formulations of the present invention are capable of preferentially delivering a payload such as a nucleic acid to cells of solid tumors compared to non-cancerous cells.
1. A nucleic acid-lipid particle comprising:
(a) a nucleic acid;
(b) a cationic lipid comprising from 45 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 cholesterol or derivative thereof comprises from 35 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 2 mol % to 15 mol % of the total lipid present in the particle, wherein the conjugated lipid comprises a polyethyleneglycol (PEG)-lipid conjugate.
2. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises an RNA.
3. The nucleic acid-lipid particle of claim 2 , wherein the RNA comprises an mRNA.
4. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 45 mol % to 60 mol % of the total lipid present in the particle.
5. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 45 mol % to 55 mol % of the total lipid present in the particle.
6. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid is protonated at a pH below the pKa of the cationic lipid.
7. The nucleic acid-lipid particle of claim 1 , wherein the mixture of the phospholipid and cholesterol or derivative thereof comprises up to 55 mol % of the total lipid present in the particle.
8. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 2 mol % to 15 mol % of the total lipid present in the particle.
9. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 2 mol % to 12 mol % of the total lipid present in the particle.
10. The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 35 mol % to 40 mol % of the total lipid present in the particle.
11. The nucleic acid-lipid particle of claim 1 , wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol, and wherein the cholesterol comprises from 35 mol % to about 40 mol % of the total lipid present in the particle.
12. The nucleic acid-lipid particle of claim 1 , wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol, and wherein the cholesterol comprises from 35 mol % to 40 mol % of the total lipid present in the particle.
13. The nucleic acid-lipid particle of claim 1 , wherein the PEG-lipid conjugate comprises from 2 mol % to 15 mol % of the total lipid present in the particle.
14. The nucleic acid-lipid particle of claim 1 , wherein the PEG-lipid conjugate comprises from 2 mol % to 12 mol % of the total lipid present in the particle.
15. The nucleic acid-lipid particle of claim 1 , wherein the PEG-lipid conjugate comprises from 3 mol % to 10 mol % of the total lipid present in the particle.
16. The nucleic acid-lipid particle of claim 1 , wherein the PEG has an average molecular weight of from about 1,000 daltons to about 5,000 daltons.
17. The nucleic acid-lipid particle of claim 1 , wherein the PEG has an average molecular weight of about 2,000 daltons.
18. The nucleic acid-lipid particle of claim 1 , wherein the PEG-lipid conjugate comprises a PEG-diacylglycerol (PEG-DAG) conjugate or a PEG-dialkyloxypropyl (PEG-DAA) conjugate.
19. A pharmaceutical composition comprising a nucleic acid-lipid particle of claim 1 and a pharmaceutically acceptable carrier.
20. The pharmaceutical composition of claim 19 , wherein the nucleic acid is fully encapsulated in the nucleic acid-lipid particle.
21. A method for introducing a nucleic acid into a cell, the method comprising:
contacting the cell with a nucleic acid-lipid particle of claim 1 .
22. A method for the in vivo delivery of a nucleic acid, the method comprising:
administering to a mammal a nucleic acid-lipid particle of claim 1 .