Ultrathin Multilayered Films for Controlled Release of Anionic Reagents

Abstract

Multilayered films, particularly ultrathin multilayered films comprising cationic polymers which are useful for controlled release of anionic species, particularly for controlled release of nucleic acids. The multilayer films herein are useful for temporal controlled released of anionic species, particularly one or more anionic peptides, proteins, nucleic acids or other anionic biological agents. In one aspect, the invention relates to multilayer films which release anionic species (anions) with separate and/or distinct release profiles, particularly wherein the anions are one or more anionic peptides, proteins or nucleic acids or other anionic biological agents

1 . A polyelectrolyte assembly comprising at least one anion/cationic polymer bilayer where the cationic polymer is a cationic polymer selected from:
 (a) a polymer of formula I:   
     
       
         
         
             
             
         
       
     
     where: 
     n, m, l, k and i are zero or integers where n+m+l+k+i=N, the total number of repeat units in the polymer; 
     A 1-3 , B 1-3 , C 1-3 , D 1-3  and E 1-3  are linkers which may be the same or each may be different; 
     Z 1 -Z 5  are most generally covalent bonds which may or may not be degradable bonds; 
     R 1-4  and R 11-19 , independently, can be hydrogen, or alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, or heteroaryl groups, all of which are optionally substituted, with the exception that R 1-4  are not esters; and 
     R 5 -R 10  are linkers or covalent bonds which may be the same or each may be different; or
 (b) a polymer of formula II: 
 
     
       
         
         
             
             
         
       
     
     where n+m=N is the number of repeating units in the polymer; 
     each y, independently, is 1, 2 or 3; each x, independently, is an integer ranging from 1-10; 
     each R 1 , each R 2 , each R 3  and each R 4 , independently, is selected from the group consisting of hydrogen, alkyl groups, alkenyl groups, alkynyl groups, carbocyclic groups, heterocyclic groups, aryl groups, heteroaryl groups, ether groups, all of which may be substituted or unsubstituted. 
   
   
       2 . The polyelectrolyte assembly of  claim 1  wherein the polymer of formula I has the formula: 
     
       
         
         
             
             
         
       
     
     where each r is an integer ranging from 1-10. 
   
   
       3 . The polyelectrolyte assembly of  claim 2  comprising one or more different anion/cationic polymer bilayers where at least a first bilayer is formed from a cationic polymer of formula I and at least a second bilayer is formed from a different cationic polymer of formula I. 
   
   
       4 . The polyelectrolyte assembly of  claim 3  wherein the cationic polymers of formula I differ from each other in the value of (m+ )/N. 
   
   
       5 . The polyelectrolyte assembly of  claim 4  wherein one of the cationic polymers of formula I has a value of (m+ )/N of 0.25 or less and the other cationic polymer of formula I has a value of (m+ )/N of 0.5 or more. 
   
   
       7 . The polyelectrolyte assembly of  claim 4  wherein one of the cationic polymers of formula I has a value of (m+ )/N of 0.25 or less and the other cationic polymer of formula I has a value of (m+ )/N of 0.75 or more. 
   
   
       8 . The polyelectrolyte assembly of  claim 4  wherein one of the cationic polymers of formula I has a value of (m+ )/N of 0.25 or less and the other cationic polymer of formula I has a value of (m+ )/N of 1.0. 
   
   
       9 . The polyelectrolyte assembly of any of  claim 2 , wherein in the cationic polymers of formula I r is an integer from 1 to 3, inclusive, and each R 3  is a hydrogen or a C1-C3 alkyl group, and each R 11 , R 12  and R 13  is a (C1-C6) alkyl group. 
   
   
       10 . The polyelectrolyte assembly of  claim 1 , further comprising one or more additional anion/cationic polymer bilayers wherein the one or more additional bilayers are formed by one or more cationic polymers of formula I wherein each cationic polymer of formula I in the assembly differs from each other cationic polymer of formula I in the assembly in the value of (m+l)/N. 
   
   
       11 . The polyelectrolyte assembly of  claim 1  which comprises at least one cationic polymer of formula I wherein the value of (m+ )/N is 0.25 or more. 
   
   
       12 . The polyelectrolyte assembly of  claim 1  which comprises at least one cationic polymer of formula I wherein the value of (m+ )/N is 0.25 or less. 
   
   
       13 . The polyelectrolyte assembly of  claim 1  which comprises at least one cationic polymer of formula I wherein the value of (m+ )/N is 0.25 or less and at least one cationic polymer of formula I wherein the value of (m+ )/N is 0.50 or more. 
   
   
       14 . The polyelectrolyte assembly of  claim 1 , which comprises only cationic polymer of formula I. 
   
   
       15 . The polyelectrolyte assembly of  claim 1  which comprises only cationic polymers of formula II. 
   
   
       16 . The polyelectrolyte assembly of  claim 15  comprising two or more different anion/cationic polymer bilayers where at least a first bilayer is formed from a cationic polymer of formula II and at least a second bilayer is formed from a different cationic polymer of formula II. 
   
   
       17 . The polyelectrolyte assembly of  claim 16  wherein the cationic polymers of formula II differ from each other in the value of (n)/(n+m). 
   
   
       18 . The polyelectrolyte assembly of  claim 17  wherein at least one of the cationic polymers of formula II has a value of (n)/(n+m) of 0.5 or more. 
   
   
       19 . The polyelectrolyte assembly of  claim 18  wherein at least one of the cationic polymers of formula II has a value of (n)/(n+m) of 0.5 or less. 
   
   
       20 . The polyelectrolyte assembly of  claim 15 , wherein in the cationic polymers of formula II, all R 2  and R 4  are hydrogens, all R 1  are C1-C3 alkyl groups and each R 3  is independently hydrogens or C1-C3 alkyl groups. 
   
   
       21 . The polyelectrolyte assembly of  claim 1  comprising two or more different anion/cationic polymer bilayers where at least a first bilayer is formed from a cationic polymer of formula I and at least a second bilayer is formed from a different cationic polymer of formula II. 
   
   
       22 . The polyelectrolyte assembly of  claim 21  wherein the at least one cationic polymer of formula I has a value of (m+ )/N is 0.50 or more. 
   
   
       23 . The polyelectrolyte assembly of  claim 21  wherein the at least one cationic polymer of formula II has a value of n/n+m of 0.5 or more. 
   
   
       24 . The polyelectrolyte assembly of  claim 23  wherein the at least one cationic polymer of formula I has a value of (m+ )/N is 0.50 or more. 
   
   
       25 . The polyelectrolyte assembly of  claim 21 , wherein in the cationic polymers of formula I, r is an integer from 1 to 3, inclusive, and each R 3  is a hydrogen or a C1-C3 alkyl group, and each R 11 , R 12  and R 13  is a (C1-C6) alkyl group. 
   
   
       26 . The polyelectrolyte assembly of  claim 21 , wherein in the cationic polymers of formula II, all R 2  and R 4  are hydrogens, all R 1  are C1-C3 alkyl groups and each R 3  is independently hydrogens or C1-C3 alkyl groups. 
   
   
       27 . The polyelectrolyte assembly of  claim 1 , wherein the anions of the bilayers are nucleic acids. 
   
   
       28 . The polyelectrolyte assembly of  claim 1 , wherein the anions of the bilayers are nucleic acids which encode polypeptides. 
   
   
       29 . The polyelectrolyte assembly of  claim 1 , wherein the polyelectrolyte assembly is formed on a substrate. 
   
   
       30 . The polyelectrolyte assembly of  claim 1 , wherein the polyelectrolyte assembly is formed on one or mroe surfaces of an implantable medical device. 
   
   
       31 . A method for controlling the release of two or more anions from a film which comprises the step of forming a polyelectrolyte assembly of  claim 1 , wherein the polyelectrolyte assembly comprises anion/cationic polymer bilayers having two or more different anions and subjecting the polyelectrolyte assembly formed to conditions in which ester functions in the polymer of formula I and in the polymer of formula II are hydrolyzed to release the two or more anions. 
   
   
       32 . The method of  claim 31  wherein the two or more anions are released from the polyelectrolyte assembly with separate, distinct or separate and distinct release profiles. 
   
   
       33 . The method of  claim 31  wherein the two or more anions are released from the polyelectrolyte assembly such that at least one anion is released short-term and at least one anion is released long-term.