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 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 2 , wherein the interfering RNA is an siRNA.
4 . The composition of claim 3 , wherein the siRNA comprises from about 15 to about 60 nucleotides.
5 . The composition of claim 3 , wherein one or more of the nucleotides in the double-stranded region of the siRNA comprise modified nucleotides.
6 . The composition of claim 5 , wherein the modified nucleotides comprise 2′-O-methyl (2′OMe) nucleotides.
7 . The composition of claim 5 , wherein less than about 50% of the nucleotides in the double-stranded region comprise modified nucleotides.
8 . The composition of claim 5 , wherein from about 20% to about 40% of the nucleotides in the double-stranded region comprise modified nucleotides.
9 . The composition of claim 1 , wherein the cationic lipid comprises 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-di-γ-linolenyloxy-N,N-dimethylaminopropane (γ-DLenDMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)[1,3]-dioxolane (DLin-K-C2-DMA), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), or a mixture thereof.
10 . The composition of claim 1 , wherein the cationic lipid comprises MC3, LenMC3, CP-LenMC3, γ-LenMC3, CP-γ-LenMC3, MC3MC, MC2MC, MC3 Ether, MC4 Ether, MC3 Amide, Pan-MC3, Pan-MC4, Pan MC5 or a mixture thereof.
11 . The composition of claim 1 , wherein the non-cationic lipid is a phospholipid.
12 . The composition of claim 1 , wherein the non-cationic lipid is cholesterol or a cholesterol derivative.
13 . The composition of claim 1 , wherein the non-cationic lipid is a mixture of a phospholipid and cholesterol or a cholesterol derivative.
14 . The composition of claim 11 , wherein the non-cationic lipid is a phospholipid selected from the group consisting of dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), or a mixture thereof.
15 . The composition of claim 11 , wherein the non-cationic lipid is DPPC.
16 . The composition of claim 1 , wherein the non-cationic lipid is a mixture of DPPC and cholesterol.
17 . The composition of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-lipid conjugate.
18 - 54 . (canceled)
55 . A method for introducing a therapeutic agent into a cell, the method comprising:
contacting the cell with a composition of claim 1 .
56 . (canceled)
57 . A method for the in vivo delivery of a therapeutic agent, the method comprising:
administering to a mammal a composition of claim 1 .
58 - 59 . (canceled)
60 . A method for treating a disease or disorder in a mammal in need thereof, the method comprising:
administering to the mammal a therapeutically effective amount of a composition of claim 1 .
61 - 62 . (canceled)