Multiblock copolymers

Abstract

Provided herein are multiblock copolymers, as well as micelles and therapeutic compositions thereof.

1 - 83 . (canceled) 
     
     
         84 . A composition comprising a block copolymer associated with a polynucleotide, the block copolymer comprising a first, a second, and a third compositionally distinct blocks, the first block being a hydrophilic polymer block, the second block being located between the first and third blocks and associated with the polynucleotide, the third block being a hydrophobic polymer block comprising anionic repeat units, the anionic repeat units having a population of anions as substituents thereof that varies in number in a pH dependant manner, the population being greater at pH 7.4 than at pH 5,
 wherein the first, second, and third blocks comprise repeat units having chain atoms and pendant groups covalently coupled to the chain atoms, and the chain atoms are carbon or a combination of carbon and oxygen atoms.   
     
     
         85 . The composition of  claim 84 , wherein at least one of the first, second, and third blocks is a random copolymer comprising two or more compositionally distinct repeat units. 
     
     
         86 . The composition of  claim 84 , wherein at least one of the first, second, and third blocks comprises repeat units of Formula 1 
       
         
           
           
               
               
           
         
         wherein 
         * designates the point of attachment of the repeat unit of Formula 1 to other repeat units; 
         each X 1  and X 2  is independently selected from the group consisting of hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclo, and substituted carbonyl, provided, however, X 1  and X 2  are not in the same repeat unit, selected from the group consisting of aryl, heteroaryl, heterosubstituted carbonyl, and combinations thereof; 
         each X 3  is independently hydrogen, alkyl or substituted alkyl, and 
         each X 4  is independently heterosubstituted carbonyl, aryl, or heteroaryl. 
       
     
     
         87 . The composition of  claim 86 , wherein at least one of the first, second, and third blocks comprises repeat units of Formula 1 in which X 4  is —C(O)OX 40 , —C(O)SX 40 , or —C(O)NX 40 X 41 , and X 40  and X 41  are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, heterohydrocarbyl, substituted heterohydrocarbyl, or heterocyclo. 
     
     
         88 . The composition of  claim 84  wherein at least one of the first and second blocks comprises repeat units derived from a polymerizable monomer having a formula 
       
         
           
           
               
               
           
         
         wherein 
         n is an integer ranging from 2 to 20, 
         X is —(CR 1 R 2 ) m — wherein m is 0-10, and wherein one or more (CR 1 R 2 ) unit is optionally substituted with —NR 1 —R 2 , —OR 1  or —SR 1 , 
         Y is —O—, —NR 4 — or —(CR 1 R 2 )—, 
         each R 1 , R 2 , R 3 , Z 1  and Z 2  are independently selected from the group consisting of hydrogen, halogen, and optionally substituted C 1 -C 3  alkyl, 
         R 4  is selected from the group consisting of hydrogen, and optionally substituted C 1 -C 6  alkyl, 
         R 8  is hydrogen or (CR 1 R 2 ) m R 9 , wherein m is 0-10, and wherein one or more (CR 1 R 2 ) unit is optionally substituted with —NR 1 R 2 , —OR 1  or —SR 1 , and 
         R 9  is hydrogen, halogen, optionally substituted C 1 -C 3  alkyl, polyol, vitamin, peptide or small molecule having a molecular weight of 200-1200 Daltons, or a conjugatable group. 
       
     
     
         89 . The composition of  claim 84 , wherein the first block comprises repeat units corresponding to Formula 1ETS: 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1ETS to other repeat units, X 3  is alkyl, and X 44  is a targeting or shielding moiety. 
     
     
         90 . The composition of  claim 84 , wherein the second block comprises repeat units corresponding to Formula 1: 
       
         
           
           
               
               
           
         
       
       X 4  is —C(O)OX 45  or —C(O)NX 45 X 41 , X 45  is substituted hydrocarbyl, substituted heterohydrocarbyl, or heterocyclo, and X 45  comprises a disulfide moiety. 
     
     
         91 . The composition of  claim 84 , wherein the third block is a hydrophobic pH-dependant, membrane destabilizing polymer, the membrane destabilizing polymer being endosome disruptive at a pH of or less than about 6.2. 
     
     
         92 . The composition of  claim 84 , wherein the third block comprises repeat units corresponding to Formula 1A: 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1A to other repeat units, and X 3  is alkyl. 
     
     
         93 . The composition of  claim 84 , wherein the third block comprises repeat units corresponding to Formula 1E: 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1E to other repeat units, and X 3  and X 47  are independently alkyl. 
     
     
         94 . The composition of  claim 84 , wherein the third block comprises repeat units corresponding to Formulae 1A and 1E 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1A and 1E to other repeat units, each X 3  and X 47  are independently alkyl, and the ratio of the number of repeat units corresponding to Formula 1A to the number of repeat units corresponding to Formula 1E in the third block is between about 20:1 and 1:4, respectively. 
     
     
         95 . The composition of  claim 84 , wherein the third block comprises repeat units corresponding to Formula 1C: 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1C to other repeat units, X 3  is alkyl, and X 48  is amino-substituted alkyl. 
     
     
         96 . The composition of  claim 84 , wherein the third block comprises repeat units corresponding to Formulae 1A and 1C 
       
         
           
           
               
               
           
         
       
       wherein * designates the point of attachment of the repeat unit of Formula 1A and 1C to other repeat units, each X 3  is independently alkyl, X 48  is amino-substituted alkyl, and the number of repeat units corresponding to Formula 1A in the third block is at least equal to the number of repeat units corresponding to Formula 1C in third block. 
     
     
         97 . The composition of  claim 84 , wherein the third block comprises anionic repeat units derived from polymerization of a (C 2 -C 8 )alkylacrylic acid. 
     
     
         98 . The composition of  claim 84 , wherein the third block further comprises a plurality of cationic repeat units derived from polymerizable monomers having a pKa ranging from 6.0 to 10.0. 
     
     
         99 . The composition of  claim 84 , wherein the polynucleotide is an siRNA, an antisense oligonucleotide, a dicer substrate, an miRNA, an aiRNA, or an shRNA. 
     
     
         100 . The composition of  claim 84 , wherein the hydrophilic first block has a structure of Formula Ia-1: 
       
         
           
           
               
               
           
         
         wherein 
         each of A 6 , A 7 , and A 8  are independently selected from the group consisting of —C—C—, —C—, —C(O)(C) a C(O)O—, —O(C) a C(O)— and —O(C) b O—, wherein a is 1-4 and b is 2-4; 
         each of Y 6 , Y 7 , and Y 8  are independently selected from the group consisting of a covalent bond, -(1C-10C)alkyl-, -(3C-6C)cycloalkyl-, —O-(1C-10C)alkyl-, —C(O)O(2C-10C)alkyl-, —OC(O)(1C-10C)alkyl-, —O(2C-10C)alkyl-, —S(2C-10C)alkyl-, —C(O)NR 6 (2C-10C)alkyl-, and aryl, any of which is optionally substituted; 
         tetravalent carbon atoms of A 6 -A 8  that are not fully substituted with R 6a , R 7 , R 8  and Y 6 -Y 8  are completed with an appropriate number of hydrogen atoms; 
         each R 6a , R 7 , and R 8  is independently selected from the group consisting of hydrogen, —CN, alkyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, any of which is optionally substituted with one or more fluorine atoms; 
         Q 6  is a residue which is hydrophilic at physiologic pH and substantially non-charged at about neutral pH; 
         Q 7  is a residue which is hydrophilic at physiologic pH and is selected from the group consisting of (i) a residue which is at least partially positively charged at about neutral pH, (ii) a residue which is at least partially negatively charged at about neutral pH but undergo protonation at lower pH, (iii) a residue which is at least partially zwitterionic at physiologic pH, and (iv) hydrogen; 
         Q 8  is selected from the group consisting of (i) a conjugatable or functionizable residue, and (ii) a targeting agent; 
         g is 0 to 1; 
         h is 0 to less than 1; 
         k is 0 to 1; wherein g+h+k=1; and 
         y is about 1 to about 100 kDa. 
       
     
     
         101 . The composition of  claim 84 , wherein the second block has a structure of Formula Ib: 
       
         
           
           
               
               
           
         
         wherein 
         A 0 , and A 1  are selected from the group consisting of —C—C—, —C—, —C(O)(C) a C(O)O—, —O(C) a C(O)— and —O(C) b O—, wherein a is 1-4 and b is 2-4; 
         Y 0 , and Y 1  are independently selected from the group consisting of a covalent bond, -(1C-10C)alkyl-, —C(O)O(2C-10C) alkyl-, —OC(O)(1C-10C) alkyl-, —O(2C-10C)alkyl-, —S(2C-10C)alkyl-, and —C(O)NR 6 (2C-10C)alkyl-; 
         tetravalent carbon atoms of A 0 -A 1  that are not fully substituted with R 1 -R 2  and Y 0 -Y 1  are completed with an appropriate number of hydrogen atoms; 
         each R 1 , R 2 , and R 6  is independently selected from the group consisting of hydrogen, —CN, alkyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, any of which is optionally substituted with one or more fluorine atoms; 
         Q 0  is a residue selected from the group consisting of residues which are hydrophilic at physiologic pH and are at least partially positively charged at physiologic pH; conjugatable or functionizable residues; and hydrogen; 
         Q 1  is a residue which is hydrophilic at physiologic pH and is selected from the group consisting of (i) a residue which is at least partially positively charged at physiologic pH, (ii) a residue which is at least partially negatively charged at physiologic pH but undergoes protonation at lower pH, (iii) a residue which is substantially neutral at physiologic pH, and (iv) a residue which is at least partially zwitterionic at physiologic pH; 
         m is 0 to less than 1.0; 
         n is greater than 0 to 1.0; wherein m+n=1; and 
         v is from about 1 to about 25 kDa. 
       
     
     
         102 . The composition of  claim 84 , wherein the third block has a structure of Formula Ic: 
       
         
           
           
               
               
           
         
         wherein 
         A 2 , A 3  and A 4  are independently selected from the group consisting of —C—C—, —C—, —C(O)(C) a C(O)O—, —O(C) a C(O)— and —O(C) b O—, wherein a is 1-4 and b is 2-4; 
         Y 4  is selected from the group consisting of hydrogen, -(1C-10C)alkyl, -(3C-6C)cycloalkyl, —O-(1C-10C)alkyl, —C(O)O(1C-10C)alkyl, —C(O)NR 6 (1C-10C) and aryl, any of which is optionally substituted with one or more fluorine groups; 
         Y 2  is selected from the group consisting of a covalent bond, -(1C-10C)alkyl-, —C(O)O(2C-10C)alkyl-, —OC(O)(1C-10C)alkyl-, —O(2C-10C)alkyl-, —S(2C-10C)alkyl-, and —C(O)NR 6 (2C-10C) alkyl-; 
         Y 3  is selected from the group consisting of a covalent bond, -(1C-10C)alkyl- and -(6C-10C)aryl-; 
         tetravalent carbon atoms of A 2 -A 4  that are not fully substituted with R 3 -R 5  and Y 2 -Y 4  are completed with an appropriate number of hydrogen atoms; 
         each R 3 , R 4 , and R 5  is independently selected from the group consisting of hydrogen, —CN, alkyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, any of which is optionally substituted with one or more fluorine atoms; 
         Q 2  is a residue which is positively charged at physiologic pH; 
         Q 3  is a residue which is negatively charged at physiologic pH, but undergoes protonation at lower pH; 
         p is about 0.1 to about 0.9; 
         q is about 0.1 to about 0.9; 
         r is 0 to about 0.8; wherein p+q+r=1; and 
         w is from about 1 to about 50 kDa. 
       
     
     
         103 . A method for intracellular delivery of a polynucleotide, the method comprising:
 contacting a cell with a composition of  claim 84 .