Amine cationic lipids and uses thereof
The present invention relates to lipid compounds and uses thereof. In particular, the compounds include a class of cationic lipids having an amine moiety, such as an amino-amine or an amino-amide moiety. The lipid compounds are useful for in vivo or in vitro delivery of one or more agents (e.g., a polyanionic payload or an antisense payload, such as an RNAi agent).
1. A compound having the formula,
wherein:
each R 1 and R 2 is, independently, C 11-24 alkyl, C 11-24 alkenyl, C 11-24 alkynyl, C 11-24 heteroalkyl, C 11-24 heteroalkenyl, or C 11-24 heteroalkynyl;
R 3 is H;
L 1 is C 1-2 alkylene; and
each R 5 and R 6 is, independently, H or C 1-6 alkyl.
2. A compound having the formula:
wherein
each R 1 and R 2 is, independently, C 11-24 alkyl, C 11-24 alkenyl, C 11-24 alkynyl, C 11-24 heteroalkyl, C 11-24 heteroalkenyl, or C 11-24 heteroalkynyl;
R 3 is H;
L 1 is C 1 alkylene; and
each R 5 and R 6 is, independently, H or C 1-6 alkyl.
3. A formulation comprising a compound of claim 1 ,
said formulation, further comprising a cationic lipid, a neutral lipid, a sterol derivative and a dsRNA.
4. A pharmaceutical composition comprising the compound of claim 1 , and a pharmaceutically acceptable excipient.
5. A method of treating or preventing a disease in a subject, said method comprising administering to said subject a compound of claim 1 , in an amount sufficient to treat said disease,
wherein said disease is selected from the group consisting of:
hepatocellular carcinoma, lung cancer, prostate cancer, or neuroblastoma.
6. A method of modulating the expression of a target nucleic acid in a subject, said method comprising administering to said subject the compound of claim 1 , in an amount sufficient to reduce the expression of said target gene in said subject,
wherein said target gene is, selected from the group consisting of ABL1, AR, β-Catenin, BCL1, BCL2, BCL6, CBFA2, CBL, CSF1R, ERBA1, ERBA2, ERBB1, ERBB2, ERBB3, ERBB4, ETS1, ETS2, ETV6, FGR, FOS, FYN, HCR, HRAS, JUN, KRAS, LCK, LYN, MET, MDM2, MLL1, MLL2, MLL3, MYB, MYC, MYCL1, MYCN, NRAS, PIM1, PML, RET, SRC, TALL TAL2, TCL3, TCL5, YES, BRCA1, BRCA2, MADH4, MCC, NF1, NF2, RB1, TP53, WT1, ApoB100, CSN5, CDK6, ITGB1, TGFβ1, Cyclin D1, PLK1, and KIF1-binding protein, and
wherein expression of said target gene is reduced in said subject.
7. A method of modulating the expression of a target nucleic acid in a subject, said method comprising administering the formulation of claim 3 in an amount sufficient to reduce the expression of said target gene in said subject,
wherein said target gene is, selected from the group consisting of ABL1, AR, β-Catenin, BCL1, BCL2, BCL6, CBFA2, CBL, CSF1R, ERBA1, ERBA2, ERBB1, ERBB2, ERBB3, ERBB4, ETS1, ETS2, ETV6, FGR, FOS, FYN, HCR, HRAS, JUN, KRAS, LCK, LYN, MET, MDM2, MLL1, MLL2, MLL3, MYB, MYC, MYCL1, MYCN, NRAS, PIM1, PML, RET, SRC, TAL1, TAL2, TCL3, TCL5, YES, BRCA1, BRCA2, MADH4, MCC, NF1, NF2, RB1, TP53, WT1, ApoB100, CSN5, CDK6, ITGB1, TGFβ1, Cyclin D1, PLK1, and KIF1-binding protein; and
wherein expression of said target gene is reduced in said subject.
8. A compound having the formula:
9. A compound having the formula:
10. The compound of claim 2 , wherein R1 is C 12 alkyl, R2 is C 18 heteroalkenyl, R3 is H, L1 is C 1 alkylene and R5 and R6 are each C 1-6 alkyl.
11. The formulation of claim 3 , wherein said cationic lipid is selected from the group consisting of N,N-dimethyl-(2,3-dioleyloxy) propylamine (DODMA), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), 1,2-dipalmitoyl-sn-glycero-O-ethyl-3-phosphocholine (DPePC), 1,2-dioleoyl-3-dimethylammonium propane (DODAP), and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP); and said neutral lipid is selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-glycero-sn-3-phosphoethanolamine (DOPE), and sphingomyelin (SM).
12. The formulation of claim 3 , wherein said cationic lipid is DODMA and said neutral lipid is DSPC.
13. The formulation of claim 3 , wherein said formulation further comprises a PEG-lipid conjugate.
14. The formulation of claim 13 , wherein said PEG-lipid conjugate is selected from the group consisting of 1,2-dimyristoyl-sn-glycerol-3-(methoxy-polyethylene glycol) (PEG-DMG), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(carbonyl-methoxy-polyethylene glycol) (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(carbonyl-methoxy-polyethylene glycol) (PEG-DSPE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(carbonyl-methoxy-polyethylene glycol) (PEG-DPPE), 1,2-dipalmitoyl-sn-glycerol-3-(methoxy-polyethylene glycol) (PEG-DPG), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(carbonyl-methoxy-polyethylene glycol) (PEG-DOPE), and 1,2-dioleoyl-sn-glycerol-3-(methoxy-polyethylene glycol) (PEG-DOG).
15. The formulation of claim 13 , wherein said PEG-lipid conjugate is PEG-DMPE or PEG-DSPE.
16. The formulation of claim 3 , wherein said sterol derivative is selected from the group consisting of cholesterol; cholestanone; cholestenone; coprostanol; 3β-[-(N—(N′, N′-dimethylaminoethane)-carbamoyl]cholesterol (DC-cholesterol); bis-guanidium-tren-cholesterol (BGTC); (2S,3 S)-2-(((3 S,10R,13R,17R)-10,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yloxy)carbonylamino)ethyl 2,3,4,4-tetrahydroxybutanoate (DPC-1); (2 S,3 S)-((3 S,10R,13R,17R)-10,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl) 2,3,4,4-tetrahydroxybutanoate (DPC-2); bis((3S,10R,13R,17R)-10,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl) 2,3,4-trihydroxypentanedioate (DPC-3); and 6-(((3 S,10R,13R,17R)-10,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yloxy)oxidophosphoryloxy)-2,3,4,5-tetrahydroxyhexanoate (DPC-4).
17. The formulation of claim 3 , wherein said sterol derivative is cholesterol.
18. The formulation of claim 13 , comprising from 20 mol % to 25 mol % of a compound of claim 1 , from 20 mol % to 30 mol % of said cationic lipid, from 2 mol % to 8 mol % of said PEG-lipid conjugate, from 10 mol % to 20 mol % of said neutral lipid, and from 25 mol % to 35 mol % of said sterol derivative.
19. The formulation of claim 15 , comprising 22 mol % of a compound of claim 1 , 26 mol % of said cationic lipid, 5 mol % to 9 mol % of said PEG-lipid conjugate, 14 mol % of said neutral lipid, and 29 mol % to 33 mol % of said sterol derivative.
20. The formulation of claim 3 , further comprising a lipid particle comprising said dsRNA and a transfection lipid.
21. The formulation of claim 13 , wherein said dsRNA comprises from 10 mol % to 40 mol % of one or more cationic lipids and from about 0.5 mol % to 10 mol % of one or more PEG-lipids.
22. The formulation of claim 20 , wherein said one or more transfection lipid, comprises from 5 mol % to 20 mol % of said neutral lipid, from 0.5 mol % to 10 mol % of said PEG-lipid conjugate, and from 20 mol % to 40 mol % of said sterol derivative.
23. The formulation of claim 3 , wherein said dsRNA has a length selected from the group consisting of 10 to 40 nucleotides, 16 to 30 nucleotides, 19 to 29 nucleotides, 25 to 35 nucleotides and 8-50 nucleotides.
24. The formulation of claim 3 , wherein said formulation, comprises from 1:10 (w/w) to about 1:100 (w/w) ratio of said dsRNA to the total lipid present in said formulation.
25. The formulation claim 3 , further comprising a liposome, a lipoplex, or a micelle.
26. The formulation of claim 25 , wherein said liposome is a lipid nanoparticle.
27. A formulation comprising a compound of claim 2 , wherein said formulation further comprises one or more components selected from a group consisting of a cationic lipid, a neutral lipid, a sterol derivative, a PEG-lipid conjugate, lipid particles comprising one or more RNA-binding agents, transfection lipids, a dsRNA, a liposome, a lipoplex, and a micelle.
28. The compound of claim 1 , wherein each R1 and R2 is, independently substituted C 11-24 alkyl, substituted C 11-24 alkenyl, substituted C 11-24 alkynyl, substituted C 11-24 heteroalkyl, substituted C 11-24 heteroalkenyl, or substituted C 11-24 heteroalkynyl.
29. The compound of claim 1 , wherein L 1 is substituted C 1-6 C 1-2 alkylene.
30. The compound of claim 1 , wherein each R 5 and R 6 is, independently, H or substituted C 1-6 alkyl.
31. The compound of claim 2 , wherein L 1 is substituted Cl alkylene.