Endosomolytic polymers
View Patent ↗We describe pH-sensitive endosomolytic polymers, delivery particles containing pH-sensitive endosomolytic polymers. The described particles are capable of delivering polynucleotides to cells from the peripheral circulation with subsequent release from endosomes. The endosomolytic polymers are inactive outside the cell but disrupt membranes upon exposure to an acidified endosomal compartment.
1. A method for delivering a polynucleotide to the cytoplasm of a cell comprising:
a) forming a first membrane active polyvinylether polymer having the formula:
wherein R1 contains an amine group, R2 is an alkyl or aryl group, and x and y are integers greater than zero;
b) forming a second membrane active polyvinylether polymer capable of causing liposome leakage having the formula:
wherein R1 contains an amine group, R2 is an alkyl or aryl group, and x and y are integers greater than zero;
c) reversibly modifying the second membrane active polyvinylether polymer via covalent linkage of a plurality of carboxydimethylmaleic anhydrides to amines on the polymer thereby forming a negatively charged reversibly inhibited membrane active polymer, wherein:
i) the reversibly inhibited membrane active polymer is not capable of causing liposome leakage, and
ii) exposure of the reversibly inhibited membrane active polymer to acidic pH results in cleavage of the disubstituted maleic anhydride from the second membrane active polyvinylether polymer;
d) associating said polynucleotide with the first amine-containing amphiphilic polyvinylether polymer to form a binary complex;
e) associating said binary complex with the reversibly inhibited membrane active polymer to form a ternary complex; and
f) contacting the cell with the ternary complex resulting in delivery of the polynucleotide to the cell.
2. The method of claim 1 wherein said first membrane active polyvinylether polymer is crosslinked to said reversibly inhibited membrane active polymer via a pH-labile bond.
3. The method of claim 1 wherein said membrane active polyvinylether polymers disrupt an endocytic membrane of the cell thereby providing delivery of said polynucleotide the cytoplasm of said cell.
4. The method of claim 1 wherein reversibly modifying the second membrane active polyvinylether polymer further comprises modification with carboxydimethylmaleic anhydride-thioester or carboxydimethylmaleic anhydride-polyethylene glycol.
5. The method of claim 4 wherein said carboxydimethylmaleic anhydrides are cleaved from said second amine-containing amphiphilic polyvinylether polymer in an endosome.
6. The method of claim 1 wherein said membrane active polyvinylether polymers each have a molecular weight greater than 10,000 Daltons.
7. The method of claim 2 wherein said ternary complex consists of a nanoparticle.
8. The method of claim 7 wherein said nanoparticle consists of a salt stable nanoparticle.
9. The method of claim 8 wherein said ternary complex has a net negative charge