Amine cationic lipids and uses thereof

Abstract

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 .- 28 . (canceled) 
     
     
         29 . A compound having the formula (IIi) or (IIj): 
       
         
           
           
               
               
           
         
         wherein each R 1  and R 2  is, independently, C 11-24  alkyl, C 11-24  alkenyl, or C 11-24  alkynyl; 
         R 3  is H or optionally substituted C 1-6  alkyl 
         L 1  is optionally substituted C 1-6  alkylene; and 
         R 5  and R 6  are independently selected from H, optionally substituted C 1-6  alkylene, C 1-6  alkyl substituted with —NR 5a R 5b  and where each R 5a  and R 5b  is, independently, H or optionally substituted C 1-6  alkyl. 
       
     
     
         30 . The compound of  claim 29 , wherein each R 1  and R 2  is C 11-24  alkenyl. 
     
     
         31 . The compound of  claim 29 , wherein the compound has formula: 
       
         
           
           
               
               
           
         
       
       and
 wherein each R 1  and R 2  is C 11-24  alkenyl. 
 
     
     
         32 . A formulation comprising a compound of  claim 29 , the formulation further comprising a cationic lipid, a neutral lipid, a sterol derivative and a dsRNA. 
     
     
         33 . The formulation of  claim 32 , wherein the 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 the 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). 
     
     
         34 . The formulation of  claim 32 , wherein the cationic lipid is DODMA and the neutral lipid is DSPC. 
     
     
         35 . The formulation of  claim 32 , wherein the formulation further comprises a PEG-lipid conjugate. 
     
     
         36 . The formulation of  claim 35 , wherein the 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). 
     
     
         37 . The formulation of  claim 35 , wherein the PEG-lipid conjugate is PEG-DMPE or PEG-DSPE. 
     
     
         38 . The formulation of  claim 32 , wherein the 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); (2S,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((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-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). 
     
     
         39 . A formulation comprising from 20 mol % to 25 mol % of the compound of  claim 29 , from 20 mol % to 30 mol % of a cationic lipid, from 2 mol % to 8 mol % of a PEG-lipid conjugate, from 10 mol % to 20 mol % of a neutral lipid, and from 25 mol % to 35 mol % of a sterol derivative. 
     
     
         40 . A formulation comprising 22 mol % of the compound of  claim 29 , 26 mol % of a cationic lipid, 5 mol % to 9 mol % of a PEG-lipid conjugate, 14 mol % of a neutral lipid, and 29 mol % to 33 mol % of a sterol derivative. 
     
     
         41 . The formulation of  claim 32 , further comprising a lipid particle comprising one or more transfection lipid(s). 
     
     
         42 . The formulation of  claim 32 , wherein the 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. 
     
     
         43 . The formulation of  claim 41 , wherein the one or more transfection lipid(s), comprises from 5 mol % to 20 mol % of the neutral lipid, from 0.5 mol % to 10 mol % of the PEG-lipid conjugate, and from 20 mol % to 40 mol % of the sterol derivative. 
     
     
         44 . The formulation of  claim 32 , wherein the 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. 
     
     
         45 . The formulation  claim 32 , further comprising a liposome, a lipoplex, or a micelle. 
     
     
         46 . The formulation of  claim 45 , wherein the liposome is a lipid nanoparticle. 
     
     
         47 . A formulation comprising a compound of  claim 29 , wherein the 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. 
     
     
         48 . A pharmaceutical composition comprising the compound of  claim 29 , and a pharmaceutically acceptable excipient. 
     
     
         49 . A method of treating or preventing a disease in a subject, the method comprising administering to the subject the compound of  claim 29 , in an amount sufficient to treat the disease, wherein the disease is selected from the group consisting of hepatocellular carcinoma, lung cancer, prostate cancer, or neuroblastoma. 
     
     
         50 . A method of modulating the expression of a target nucleic acid in a subject, the method comprising administering to the subject the compound of  claim 29 , in an amount sufficient to reduce the expression of the target gene in the subject,
 wherein the 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 the target gene is reduced in the subject.   
     
     
         51 . The compound of  claim 29 , wherein the compound is selected from L-37, L-38, L-39, L-40, L-41, L-42 and L-43. 
     
     
         52 . The compound of  claim 29 , wherein each R 1  and R 2  is, independently, selected from the group consisting of linolenyl (C18:3), linolenyloxy (C18:3), linolenoyl (C18:3), linoleyl (C18:2), linoleyloxy (C18:2), linoleoyl (C18:2), oleyl (C18:1), oleyloxy (18:1), oleyloxymethylene (18:1), oleoyl (C18:1), oleoylmethylene (C18:1), stearyl (C18:0), stearyloxy (C18:0), stearoyl (C18:0), palmityl (C16:0), palmityloxy (C16:0), palmitoyl (C16:0), palmitoylmethylene (C16:0), myristyl (C14:0), myristyloxy (C14:0), myristoyl (C14:0), lauryl (C12:0), lauryloxy (C12:0), and lauroyl (C12:0). 
     
     
         53 . The compound of  claim 52 , wherein each R 1  and R 2  is, independently, linoleyl (C18:2) or oleyl (C18:1).