NOVEL 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 - 64 . (canceled)
65 . 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 from 20 mol % to 55 mol % of the total lipid present in the particle, wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol or a derivative thereof; and
(d) a conjugated lipid that inhibits aggregation of particles comprising from 2 mol % to 15 mol % of the total lipid present in the particle, wherein the conjugated lipid comprises a polyethyleneglycol (PEG)-lipid conjugate,
wherein the particle has a size of from 50 nm to 60 nm.
66 . The nucleic acid-lipid particle of claim 65 , wherein the nucleic acid comprises an RNA.
67 . The nucleic acid-lipid particle of claim 66 , wherein the RNA comprises an mRNA.
68 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 45 mol % to 60 mol % of the total lipid present in the particle.
69 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 50 mol % to 65 mol % of the total lipid present in the particle.
70 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 50 mol % to 60 mol % of the total lipid present in the particle.
71 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 52 mol % to 58 mol % of the total lipid present in the particle.
72 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 52 mol % to 56 mol % of the total lipid present in the particle.
73 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid comprises from 53 mol % to 55 mol % of the total lipid present in the particle.
74 . The nucleic acid-lipid particle of claim 65 , wherein the cationic lipid is protonated at a pH below the pKa of the cationic lipid.
75 . The nucleic acid-lipid particle of claim 65 , wherein the mixture of the phospholipid and cholesterol or derivative thereof comprises from 30 mol % to 50 mol % of the total lipid present in the particle.
76 . The nucleic acid-lipid particle of claim 65 , wherein the mixture of the phospholipid and cholesterol or derivative thereof comprises from 35 mol % to 45 mol % of the total lipid present in the particle.
77 . The nucleic acid-lipid particle of claim 65 , wherein the phospholipid comprises from 2 mol % to 15 mol % of the total lipid present in the particle.
78 . The nucleic acid-lipid particle of claim 65 , wherein the phospholipid comprises from 2 mol % to 12 mol % of the total lipid present in the particle.
79 . The nucleic acid-lipid particle of claim 65 , wherein the phospholipid comprises from 4 mol % to 15 mol % of the total lipid present in the particle.
80 . The nucleic acid-lipid particle of claim 65 , wherein the cholesterol or derivative thereof comprises from 25 mol % to 45 mol % of the total lipid present in the particle.
81 . The nucleic acid-lipid particle of claim 65 , wherein the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle.
82 . The nucleic acid-lipid particle of claim 65 , wherein the cholesterol or derivative thereof comprises from 30 mol % to 35 mol % of the total lipid present in the particle.
83 . The nucleic acid-lipid particle of claim 65 , wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol, and wherein the cholesterol comprises from 25 mol % to 45 mol % of the total lipid present in the particle.
84 . The nucleic acid-lipid particle of claim 65 , wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol, and wherein the cholesterol comprises from 30 mol % to 40 mol % of the total lipid present in the particle.
85 . The nucleic acid-lipid particle of claim 65 , wherein the non-cationic lipid comprises a mixture of a phospholipid and cholesterol, and wherein the cholesterol comprises from 30 mol % to 35 mol % of the total lipid present in the particle.
86 . The nucleic acid-lipid particle of claim 65 , wherein the PEG-lipid conjugate comprises from 2 mol % to 12 mol % of the total lipid present in the particle.
87 . The nucleic acid-lipid particle of claim 65 , wherein the PEG has an average molecular weight of from about 1,000 daltons to about 5,000 daltons.
88 . The nucleic acid-lipid particle of claim 65 , wherein the PEG has an average molecular weight of about 2,000 daltons.
89 . The nucleic acid-lipid particle of claim 65 , wherein the PEG-lipid conjugate comprises a PEG-diacylglycerol (PEG-DAG) conjugate or a PEG-dialkyloxypropyl (PEG-DAA) conjugate.
90 . The nucleic acid-lipid particle of claim 65 , wherein the particle has a size of about 55 nm.
91 . The nucleic acid-lipid particle of claim 65 , wherein the particle is substantially non-toxic.
92 . A pharmaceutical composition comprising a nucleic acid-lipid particle of claim 65 and a pharmaceutically acceptable carrier.
93 . The pharmaceutical composition of claim 92 , wherein the nucleic acid is fully encapsulated in the nucleic acid-lipid particle.
94 . A method for introducing a nucleic acid into a cell, the method comprising:
contacting the cell with a nucleic acid-lipid particle of claim 65 .
95 . 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 65 .
96 . The method of claim 95 , wherein the mammal is a human.