Enhanced transport using membrane disruptive agents

Abstract

Compositions and methods for transport or release of therapeutic and diagnostic agents or metabolites or other analytes from cells, compartments within cells, or through cell layers or barriers are described. The compositions include a membrane barrier transport enhancing agent and are usually administered in combination with an enhancer and/or exposure to stimuli to effect disruption or altered permeability, transport or release. In a preferred embodiment, the compositions include compounds which disrupt endosomal membranes in response to the low pH in the endosomes but which are relatively inactive toward cell membranes, coupled directly or indirectly to a therapeutic or diagnostic agent. Other disruptive agents can also be used, responsive to stimuli and/or enhancers other than pH, such as light, electrical stimuli, electromagnetic stimuli, ultrasound, temperature, or combinations thereof. The compounds can be coupled by ionic, covalent or H bonds to an agent to be delivered or to a ligand which forms a complex with the agent to be delivered. Agents to be delivered can be therapeutic and/or diagnostic agents. Treatments which enhance delivery such as ultrasound, iontophoresis, and/or electrophoresis can also be used with the disrupting agents.

1 - 8 . (canceled) 
     
     
         9 . A composition for delivering a therapeutic agent to a cell, comprising:
 (a) a transport agent; and   (b) a peptide covalently coupled to the transport agent,   wherein the transport agent is hydrophobic at a pH from 5.1 to 5.5 and effective in disrupting an endosomal membrane, and the transport agent is a graft copolymer or block copolymer comprising one or more propylacrylic acid groups.   
     
     
         10 . The composition of  claim 9 , wherein the transport agent is a block copolymer. 
     
     
         11 . The composition of  claim 9 , further comprising an oligonucleotide. 
     
     
         12 . The composition of  claim 11 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         13 . The composition of  claim 12 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         14 . The composition of  claim 11 , wherein the oligonucleotide is an RNA. 
     
     
         15 . The composition of  claim 9 , wherein the peptide is GALA or mellitin. 
     
     
         16 . The composition of  claim 9 , wherein the peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         17 . The composition of  claim 9 , wherein the peptide is polylysine. 
     
     
         18 . A composition for delivering a therapeutic agent to a cell, comprising:
 (a) a transport agent; and   (b) a peptide covalently coupled to the transport agent,   wherein the transport agent is hydrophobic at a pH from 5.1 to 5.5 and effective in disrupting an endosomal membrane, the transport agent is a random copolymer comprising one or more propylacrylic acid groups.   
     
     
         19 . The composition of  claim 18 , further comprising an oligonucleotide. 
     
     
         20 . The composition of  claim 19 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         21 . The composition of  claim 20 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         22 . The composition of  claim 19 , wherein the oligonucleotide is an RNA. 
     
     
         23 . The composition of  claim 18 , wherein the peptide is GALA or mellitin. 
     
     
         24 . The composition of  claim 18 , wherein the peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         25 . The composition of  claim 18  wherein the peptide is polylysine. 
     
     
         26 . A pharmaceutical composition for delivering a therapeutic agent to a cell, comprising:
 (a) a transport agent; and   (b) a therapeutic agent,   wherein the transport agent is hydrophobic at a pH from 5.1 to 5.5 and effective in disrupting an endosomal membrane, the transport agent is a random copolymer comprising one or more propylacrylic acid groups and at least one covalently-coupled peptide.   
     
     
         27 . The pharmaceutical composition of  claim 26 , wherein the therapeutic agent is an oligonucleotide. 
     
     
         28 . The pharmaceutical composition of  claim 27 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         30 . The pharmaceutical composition of  claim 26 , wherein the therapeutic agent is an RNA. 
     
     
         31 . The pharmaceutical composition of  claim 26 , wherein the at least one peptide comprises GALA or mellitin. 
     
     
         32 . The pharmaceutical composition of  claim 26  wherein the at least one peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         33 . The pharmaceutical composition of  claim 26 , wherein the at least one peptide comprises polylysine. 
     
     
         34 . A method for delivering a therapeutic agent to a cell, comprising contacting the cell with a composition of  claim 9 . 
     
     
         35 . The method of  claim 34 , wherein the composition further comprises an oligonucleotide. 
     
     
         36 . The method of  claim 35 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         37 . The method of  claim 36 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         38 . The method of  claim 35 , wherein the oligonucleotide is an RNA. 
     
     
         39 . The method of  claim 34 , wherein the peptide is GALA or mellitin. 
     
     
         40 . The method of  claim 34 , wherein the peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         41 . The method of  claim 34 , wherein the peptide is polylysine. 
     
     
         42 . A method for delivering a therapeutic agent to a cell, comprising contacting the cell with a composition of  claim 18 . 
     
     
         43 . The method of  claim 42 , wherein the composition further comprises an oligonucleotide. 
     
     
         44 . The method of  claim 43 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         45 . The method of  claim 44 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         46 . The method of  claim 43 , wherein the oligonucleotide is an RNA. 
     
     
         47 . The method of  claim 42 , wherein the peptide is GALA or mellitin. 
     
     
         48 . The method of  claim 42 , wherein the peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         49 . The method of  claim 42 , wherein the peptide is polylysine. 
     
     
         50 . A method for delivering a therapeutic agent to a cell, comprising contacting the cell with a pharmaceutical composition of  claim 26 . 
     
     
         51 . The method of  claim 50 , wherein the therapeutic agent is an oligonucleotide. 
     
     
         52 . The method of  claim 51 , wherein the oligonucleotide is a modified oligonucleotide. 
     
     
         53 . The method of  claim 52 , wherein the modified oligonucleotide comprises modified purine bases, modified pyrimidine bases, phosphorothioate linkages, or any combination thereof. 
     
     
         54 . The method of  claim 51 , wherein the oligonucleotide agent is an RNA. 
     
     
         55 . The method of  claim 50 , wherein the peptide is GALA or mellitin. 
     
     
         56 . The method of  claim 50 , wherein the peptide consists of a repeating glutamic acid-alanine-leucine-alanine structure. 
     
     
         57 . The method of  claim 50 , wherein the peptide is polylysine. 
     
     
         58 . A method for delivering a therapeutic agent to a cell, comprising:
 (a) contacting a cell with a composition comprising:
 (i) a transport agent effective in disrupting the endosomal membrane; and 
 (ii) a peptide covalently coupled to the transport agent, 
 wherein the transport agent and peptide are taken into the cell by endocytosis to provide an endosome having an endosomal membrane and containing the transport agent and peptide, and 
 wherein the transport agent is a graft copolymer or block copolymer comprising one or more acrylic acid groups or one or more alkyl-substituted acrylic acid groups, provided that the graft copolymer or block copolymer comprises one or more propylacrylic acid groups or one or more butylacrylic acid groups; and 
   (b) releasing the composition from the endosome into the cell cytoplasm by action of the transport agent on the endosomal membrane.