Heterogeneous polymeric micelles for intracellular delivery

1. A composition comprising a heterogeneous polymeric micelle and a polynucleotide associated with the micelle, the micelle comprising:

a first polymer, the first polymer being a block copolymer comprising a hydrophilic block and a hydrophobic block; and

a second polymer compositionally distinct from the first polymer, the second polymer being a block copolymer comprising a hydrophilic block and a hydrophobic block, the hydrophobic block of the second polymer being associated with the hydrophobic block of the first polymer such that the micelle is stable in an aqueous medium at pH 7.4,

wherein at least one of the first polymer or the second polymer comprises a pH-dependent, membrane-destabilizing polymer;

wherein the hydrophilic block of the first polymer comprises a plurality of cationic monomeric residues in ionic association with the polynucleotide,

wherein the hydrophilic block of the second polymer is a neutral hydrophilic block comprising a plurality of neutral polar residues, and

wherein the first polymer or the second polymer is a block copolymer of Formula IA:

wherein

A 0 , A 1 , A 2 , A 3 , and A 4 are each independently selected from the group consisting of —CH 2 —, —C—C—, —C(O)(C) a C(O)O—, —O(C) a C(O)— and —O(C) b O—;

a is an independently selected integer ranging from 1-4;

b is an independently selected integer ranging from 2-4;

m ranges from 0 to less than 1.0;

n ranges from greater than 0 to about 1.0;

the sum of (m+n)=1;

p ranges from about 0.1 to about 0.9;

q ranges from about 0.1 to about 0.9;

r ranges from 0 to about 0.8;

the sum of (p+q+r)=1;

Y 0 , Y 1 , and Y 2 are each 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-;

Y 3 is selected from the group consisting of a covalent bond, (1C-10C) alkyl and (6C-10C) aryl;

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;

tetravalent carbon atoms of A 0 -A 4 that are not fully substituted with R 1 -R 5 ; and

Y 0 -Y 4 are substituted with a valency-appropriate number of hydrogen atoms;

R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are independently selected from the group consisting of hydrogen, —CN, alkyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, any of which may be optionally substituted with one or more fluorine atoms;

Q 0 is selected from the group consisting of hydrogen, hydrophilic species (at physiologic pH), L conjugatable species and functional species;

Q 1 is a species which is hydrophilic at physiologic pH;

Q 2 is a species which is positively charged at physiologic pH;

Q 3 is a species which is negatively charged at physiologic pH, but undergoes protonation at lower pH;

v corresponds to a molecular weight from about 5 to about 25 kDa; and,

w corresponds to a molecular weight from about 5 to about 50 kDa.

2. The composition of claim 1 , wherein the polynucleotide is an siRNA.

3. A method for intracellular delivery of a polynucleotide, the method comprising:

contacting a composition of claim 1 with a cell surface in a medium at a first pH;

introducing the composition into an endosomal membrane within the cell through endocytosis; and

destabilizing the endosomal membrane, whereby the composition or the polynucleotide is delivered to the cytosol of the cell.

4. The composition of claim 1 , wherein the hydrophobic block of the second polymer is compositionally distinct from the hydrophobic block of the first polymer.

5. The composition of claim 1 , wherein the hydrophobic block of the second polymer has substantially the same composition as the hydrophobic block of the first polymer.

6. The composition of claim 1 , wherein the second polymer comprises one or more monomeric residues that are different than the monomeric residues of the first polymer.

7. The composition of claim 1 , further comprising a targeting moiety.

8. The composition of claim 1 , comprising a targeting moiety, wherein the targeting moiety is a ligand having affinity for one or more receptors effective for mediating endocytosis.

9. The composition of claim 1 , comprising a targeting moiety, wherein the targeting moiety is covalently coupled to a hydrophilic block of the first polymer or to a hydrophilic block of the second polymer.