Gene delivery carrier

Abstract

Disclosed are methods, compositions, reagents, systems, and kits to prepare and utilize poly (β-amino ester) (PBAE) polymers, which are synthesized via Michael addition reactions of diacrylates and amines disclosed herein. Various embodiments utilize lactones and lactone derivatives to generate the diacrylate compounds. The PBAE polymers are shown to be effective biodegradable carriers for the delivery of an agent such as an organic molecule, inorganic molecule, nucleic acid, protein, peptide, polynucleotide, targeting agent, an isotopically labeled chemical compound, vaccine, or an immunological agent.

1 . A polymer of Formula (I): 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof, 
         wherein:
    is a single bond or a double bond 
 A is hydrogen or —XR 3 ; 
 each B independently is a diradical of the formula: 
 
       
       
         
           
           
               
               
           
         
         
           each D independently is: 
         
       
       
         
           
           
               
               
           
         
         
           E is hydrogen or —XR 4 ; 
           each R, R′, and R″ independently is optionally substituted, cyclic or acyclic aliphatic; optionally substituted, cyclic or acyclic heteroaliphatic; or a combination thereof; 
           each R a  independently is hydrogen or optionally substituted aliphatic; 
           R 1  and R 2  independently are selected from hydrogen, optionally substituted, cyclic or acyclic aliphatic; optionally substituted, cyclic or acyclic heteroaliphatic; optionally substituted aryl; optionally substituted heteroaryl; and a nitrogen protecting group; or R 1  and R 2  are combined to form a ring; 
           R 3  and R 4  independently are selected from hydrogen; optionally substituted, cyclic or acyclic aliphatic; optionally substituted, cyclic or acyclic heteroaliphatic; optionally substituted aryl; optionally substituted heteroaryl; and a protecting group; 
           X is O, S, NH, or NR X , wherein R X  is optionally substituted, aliphatic; optionally substituted carbocyclyl; optionally substituted heteroaliphatic; optionally substituted heterocyclyl; optionally substituted aryl; or optionally substituted heteroaryl; 
           X′ is O or NR y , wherein R y  is hydrogen; optionally substituted, aliphatic; optionally substituted carbocyclyl; optionally substituted heteroaliphatic; optionally substituted heterocyclyl; optionally substituted aryl; or optionally substituted heteroaryl; 
           each m independently is an integer between 1 and 100, inclusive; and 
           n is an integer between 1 and 10,000, inclusive. 
         
       
     
     
         2 .- 4 . (canceled) 
     
     
         5 . The polymer of  claim 1 , having the structure of Formula (III) or Formula (V): 
       
         
           
           
               
               
           
         
       
     
     
         6 .- 35 . (canceled) 
     
     
         36 . The polymer of  claim 1 , wherein B is of the formula: 
       
         
           
           
               
               
           
         
       
     
     
         37 . (canceled) 
     
     
         38 . The polymer of  claim 1 , wherein m is an integer between 3 and 7, inclusive 
     
     
         39 .- 85 . (canceled) 
     
     
         86 . The polymer of  claim 1 , wherein the polymer is of the formula: 
       
         
           
           
               
               
           
         
         wherein:
 each D is 
 
       
       
         
           
           
               
               
           
         
       
     
     
         87 .- 93 . (canceled) 
     
     
         94 . The polymer of  claim 1 , wherein the polymer has a molecular weight between 1-100 kDa. 
     
     
         95 .- 113 . (canceled) 
     
     
         114 . A method of preparing a polymer of Formula (III), as defined in  claim 5 , or a pharmaceutically acceptable salt thereof: 
       
         
           
           
               
               
           
         
         comprising reacting a polymer of Formula (V), as defined in  claim 1 , or a pharmaceutically acceptable salt thereof: 
       
       
         
           
           
               
               
           
         
         with one or more nucleophiles selected from R 3 XH and R 4 XH; to obtain the polymer of Formula (III). 
       
     
     
         115 .- 117 . (canceled) 
     
     
         118 . A composition comprising a polymer of  claim 1  and an excipient. 
     
     
         119 .- 123 . (canceled) 
     
     
         124 . The composition of  claim 118 , wherein the composition further comprises an agent. 
     
     
         125 . (canceled) 
     
     
         126 . The composition of  claim 124 , wherein the agent is a polynucleotide. 
     
     
         127 . The composition of  claim 126 , wherein the polynucleotide is DNA. 
     
     
         128 . The composition of  claim 126 , wherein the polynucleotide is RNA. 
     
     
         129 . The composition of  claim 128 , wherein the RNA is dsRNA, siRNA, shRNA, miRNA, mRNA, or antisense RNA. 
     
     
         130 . The composition of  claim 126 , wherein the polynucleotide carries out RNA interference. 
     
     
         131 . (canceled) 
     
     
         132 . The composition of  claim 118 , wherein the composition is in the form of a particle. 
     
     
         133 .- 134 . (canceled) 
     
     
         135 . The composition of  claim 132 , wherein the particle encapsulates an agent. 
     
     
         136 . A method of treating a disease, disorder, or condition from which a subject suffers, comprising administering to a subject in need thereof an effective amount of a composition of  claim 118 . 
     
     
         137 . (canceled) 
     
     
         138 . A method of delivering a polynucleotide to a cell, comprising contacting the cell with a composition of  claim 126 . 
     
     
         139 .- 140 . (canceled) 
     
     
         141 . The composition of  claim 128 , wherein the RNA is mRNA. 
     
     
         142 . The composition of  claim 141 , wherein the mRNA encodes an antigen.