Lipopeptides for delivery of nucleic acids

Abstract

Lipopeptide compounds comprising a peptide having 2 to 100 amino acid residues, and having a lipophilic group attached to at least one terminus of the peptide or to at least one amino acid residue of the peptide, and salts and uses thereof. The lipophilic group may be attached to the N-terminus, C-terminus or both termini of the peptide. The lipophilic group may be attached to at least one interal amino acid residue (i.e., an amino acid residue that is not the N-terminus or the C-terminus amino acid residue of the peptide). The lipophilic group may be attached to either termini or both and at least one internal amino acid residue.

1 . The compound of  claim 1 , comprising the structure shown in Formula I:
   R 3 -J-{NR N —CR 1 R 2 —(C═O)} n —Z—R 4   Formula I
   wherein
 R 1  is independently, for each occurrence, a non-hydrogen side chain of an amino acid; 
 {NR N —CR 1 R 2 —(C═O)} n  is a central peptide; 
   R 2  and R N  are independently of one another hydrogen, or an organic group consisting of carbon, oxygen, nitrogen, sulfur, and hydrogen atoms, and having from 1 to 20 carbon atoms, or C(1-22)alkyl, (6-12)cycloalkyl, (6-12)cycloalkylalkyl, C(3-18)alkenyl, C(3-18)alkynyl, C(1-5)alkanoyl, C(1-5)alkanoyloxy, C(1-5)alkoxy, C(1-5)alkoxy-C(1-5)alkyl, C(1-5) alkoxy-C(1-5)alkoxy, C(1-5)alkyl-amino-C(1-5)alkyl-, C(1-5)dialkyl-amino-C(1-5)alkyl-, nitro-C(1-5)alkyl, cyano-C(1-5)alkyl, aryl-C(1-5)alkyl, 4-biphenyl-C(1-5)alkyl, carboxyl, or hydroxyl;   R 3  and R 4  are independently of one another, a lipophilic tail derived from a naturally-occurring or synthetic lipid, phospholipid, glycolipid, triacylglycerol, glycerophospholipid, sphingolipid, ceramide, sphingomyelin, cerebroside, or ganglioside, wherein the tail may contain a steroid, or an organic group consisting of carbon, oxygen, nitrogen, sulfur, and hydrogen atoms, and having from 1 to 20 carbon atoms, or a substituted or unsubstituted C(1-22)alkyl, C(6-12)cycloalkyl, C(6-12)cycloalkyl-alkyl, C(3-18)alkenyl, C(3-18)alkynyl, C(1-5)alkoxy-C(1-5)alkyl, or a sphinganine, (2R,3R)-2-amino-1,3-octadecanediol, icosasphinganine, sphingosine, phytosphingosine, cis-4-sphingenine, or a ceramide;   Z is NH, O, or a linker comprising a maleimido, thioether, amide, cysteamide, cysteine, thiol, or a disulfide group, or a polyethyleneoxide or polypropyleneoxide group comprising 1-400 atoms, or a linker comprising 1-200 atoms selected from the group of C, H, F, Cl, Br, N, O, S, Si, and P;   J is (C═O), O, or a linker comprising a maleimido, thioether, amide, cysteamide, cysteine, thiol, or disulfide group, or a polyethyleneoxide or polypropyleneoxide group comprising 1-400 atoms, or a linker comprising 1-200 atoms selected from the group of C, H, F, Cl, Br, N, O, S, Si, and P; and   
       wherein
 n is 2 to 100. 
 
     
     
         2 . The compound of  claim 1 , wherein the amino acid sequence of the central peptide is selected from the group consisting of SEQ ID NOs: 1-300 and 350-356. 
     
     
         3 . The compound of  claim 1 , wherein the amino acid sequence of the central peptide is selected from the group consisting of GALFLAFLAAALX aa LMGLWX aa QX aa KKKRKV (SEQ ID NO: 291), WSQPKKKRKV (SEQ ID NO: 292), WWHHKKRRCCRRKKHHWW (SEQ ID NO:94), X aa WSQPKKKRKVX aa  (SEQ ID NO:293), WX aa QPKKKRKX aa  (SEQ ID NO:294) X aa X aa QPKKKRKV(SEQ ID NO:295), LIRLWX aa HLIHIWFQX aa RRLKWKKK (SEQ ID NO: 297), QNRRLKWKKK (SEQ ID NO:298), X aa QNRRLKWKKKX aa  (SEQ ID NO:299), and QX aa RRLKWKKK (SEQ ID NO:300), where X aa  is independently, for each occurance, L, A, Q, H, E, R, or K. 
     
     
         4 . The compound of  claim 1 , wherein a side chain is linked to the central peptide, wherein the side chain contains a releasing functional group selected from 3,5-diiodo-tyrosine, 1-methylhistidine, 2-methylbutanoic acid, 2-o-anisylpropanoic acid, meso-tartaric acid, 4,6-dimethylpyrimidinamine, p-phthalic acid, creatinine, butanoic acid, N,N-dimethyl-1-naphthylamine, pentanoic acid, 4-methylpentanoic acid, N-methylaniline, 1,10-phenanthroline, 3-pyridinecarboxylic acid, hexanoic acid, propanoic acid, 4-animobenzoic acid, 2-methylpropanoic acid, heptanoic acid, octanoic acid, cyclohexanecarboxylic acid, quinoline, 3-quinolinamine, 2-aminobenzoic acid, 4-pyridinecarboxylic acid, nonanic acid, melamine, 8-quinolinol, trimethylacetic acid, 6-methoxyquinoline, 4-(methylamino)benzoic acid, p-methylaniline, 3-(methylamino)benzoic acid, malic acid, N-ethylaniline, 2-benzylpyridine, 3,6-dinitrophenol, N,N-dimethylaniline, 2,5-dimethylpiperazine, p-phenetidine, 5-methylquinoline, 2-phenylbenzimidazole, pyridine, picolinic acid, 3,5-diiodityrosine, p-anisidine, 2-(methylamino)benzoic acid, 2-thiazolamine, glutaric acid, adipic acid, isoquinoline, itaconic acid, o-phthalic acid, benzimidazole, piperazine, heptanedioic acid, acridine, phenanthridine, succinic acid, methylsuccinic acid, 4-methylquinoline, 3-methylpyridine, 7-isoquinolinol, malonic acid, methymalonic acid, 2-methylquinoline, 2-ethylpyridine, 2-methylpyridine, 4-methylpyridine, histamine, histidine, maleic acid, cis-1,2-cyclohexanediamine, 3,5-dimethylpyridine, 2-ethylbenzimidazole, 2-methylbenzimidazole, cacodylic acid, perimidine, citric acid, isocitric acid, 2,5-dimethylpyridine, papaverine, 6-hydroxy-4-methylpteridine, L-thyroxine, 3,4-dimethylpyridine, methoxypyridine, trans-1,2-cyclohexanediamine, 2,5-pyridinediamine, l-1-methylhistidine, l-3-methylhistidine, 2,3-dimethylpyridine, xanthopterin, 1,2-propanediamine, N,N-diethylaniline, alloxanic acid, 2,6-dimethylpyridine, L-carnosine, 2-pyridinamine, N-b-alanylhistidine, pilocarpine, 1-methylimidazol, 1H-imidazole, 2,4-dimethylpyridine, 4-nitrophenol, 2-nitrophenol, tyrosineamide, 5-hydroxxyquinazoline, 1,1-cyclopropanedicarboxylic acid, 2,4,6-trimethylpyridine, veronal, 2,3-dichlorophenol, 1,2-ethanediamine, 1-isoquinolinamine, and combinations thereof. 
     
     
         5 . A pharmaceutical composition comprising a nucleic acid agent and one or more compounds according to  claim 1 , or pharmaceutically-acceptable salts thereof. 
     
     
         6 . The composition of  claim 5 , wherein the nucleic acid agent is a usiRNA. 
     
     
         7 . The composition of  claim 5 , wherein the nucleic acid agent is an mdRNA. 
     
     
         8 . The composition of  claim 5 , wherein the nucleic acid agent is an siRNA. 
     
     
         9 . The composition of  claim 5 , wherein the one or more compounds is (C20)-(H) 8 (R) 8 (H) 8 K-NH-(C20) (SEQ ID NO: 13). 
     
     
         10 . The composition of  claim 5 , further comprising a non-cationic lipid. 
     
     
         11 . The composition of  claim 5 , further comprising a polymeric lipid. 
     
     
         12 . A method for delivering an interfering RNA agent to a cell comprising preparing a composition according to  claim 5  and treating a cell with the composition. 
     
     
         13 . A method for inhibiting expression of a gene in a cell comprising preparing a composition according to  claim 5  and treating a cell with the composition. 
     
     
         14 . A method for inhibiting expression of a gene in a mammal comprising preparing a composition according to  claim 5  and administering the composition to the mammal. 
     
     
         15 . A method for treating the signs and symptoms of a disease, disorder, or condition in a subject selected from cancer, a proliferative disease, disorder, or condition, a metabolic disease, disorder, or condition, an inflammatory disease, disorder, or condition, and a viral infection by providing a composition of  claim 5  and administering the composition to the subject.