Therapeutic cationic peptides and unimolecular nanoparticles for efficient delivery thereof
Provided herein are peptides comprising an amino acid sequence having at least about 85% sequence identity to RYRPRAPIIAVT (SEQ ID NO: 1). These cationic peptides inhibit PKM2 methylation and may be used in the treatment of breast cancer and other diseases or conditions in which PKM2 is overexpressed. Such PKM2 peptides may be delivered to cancer cells using pH sensitive unimolecular nanoparticles comprising anionic polymers.
1. A unimolecular nanoparticle comprising:
a dendritic polymer core having a molecular weight of 500 to 120,000 Da and terminating in hydroxyl, amino or carboxylic acid groups;
anionic polymers attached to at least a majority of the terminating groups of the dendritic polymer, wherein
each anionic polymer comprises a polymeric backbone attached to weakly basic groups by a C 2 -C 12 heteroalkyl group comprising 1 to 4 nitrogen atoms and anionic functional groups, wherein
the anionic functional groups are each conjugated to the polymeric backbone via a pH sensitive linker and optionally the C 2 -C 12 heteroalkyl group,
the number of anionic functional groups per repeat unit of the anionic polymer ranges from 1-2 and the number of weakly basic groups per repeat unit of the anionic polymer ranges from 0 to 2, and
each anionic polymer has a molecular weight from about 1,000 to about 10,000 Da; and
poly(ethylene glycol) attached to a plurality of anionic polymers and having a terminal group selected from a targeting ligand, OH, O-alkyl, NH 2 , biotin, or a dye, wherein the poly(ethylene glycol) has a molecular weight of about 1,000 to about 15,000 Da.
2. The unimolecular nanoparticle of claim 1 , wherein the dendritic polymer has from 3-7 generations.
3. The unimolecular nanoparticle of claim 2 , wherein the dendritic polymer has 3 or 4 generations.
4. The unimolecular nanoparticle of claim 1 wherein the pH-sensitive linker comprises an imine, hydrazone, cis-aconityl, acetal, β-thiopropionate group, or a combination of two or more thereof.
5. The unimolecular nanoparticle of claim 1 wherein the pH-sensitive linker comprises an imine, cis-aconityl, or a combination thereof.
6. The unimolecular nanoparticle of claim 1 wherein the anionic polymer comprises a polyamide backbone, alkylene-amino-alkylene linker, imidazole groups, pH: sensitive linker, and anionic functional groups selected from carboxyl, sulfonic or phosphate groups.
7. The unimolecular nanoparticle of claim 6 wherein the polyamide backbone comprises polyasparagine, polyglutamine, polyornithine, or polylysines.
8. The unimolecular nanoparticle of claim 1 wherein each anionic polymer has a molecular weight from about 1,000 to about 5,000 Da.
9. The unimolecular nanoparticle of claim 1 wherein the targeting ligand is a cofactor, carbohydrate, peptide, antibody, nanobody, or aptamer.
10. The unimolecular nanoparticle of claim 1 wherein the molar ratio of the anionic functional groups to the weakly basic groups ranges from 1:1 to 10:1.
11. The unimolecular nanoparticle of claim 1 further comprising a therapeutic cationic peptide within the nanoparticle.
12. The unimolecular nanoparticle of claim 11 wherein the therapeutic cationic peptide is bound by electrostatic interactions with the anionic polymers.
13. The unimolecular nanoparticle of claim 11 wherein the loading of the therapeutic cationic peptide is about 1 to about 20 wt % of the unimolecular nanoparticle.
14. The unimolecular nanoparticle of claim 1 wherein
the dendritic polymer is a PAMAM dendrimer having 3-4 generations and a molecular weight of about 5,000 to about 15,000 Da;
the pH-sensitive linker is a cis-aconityl group;
each anionic polymer has a polyasparagine backbone attached to an ethylene-amino-ethyleneamino-cis-aconityl and ethylene-amino-ethylamidoimidazole group or salt thereof and the ratio of the carboxyl groups to imidazole groups is from 1:1 to 5:1; and
the molecular weight of the PEG is about 1,000 to about 15,000 Da.
15. The unimolecular nanoparticle of claim 14 further comprising a therapeutic cationic peptide within the nanoparticle.
16. A composition comprising the unimolecular nanoparticle of claim 1 and a pharmaceutically acceptable carrier.
17. A composition comprising the unimolecular nanoparticle of claim 11 and a pharmaceutically acceptable carrier.
18. A kit comprising a package containing unimolecular nanoparticle of claim 1 and a package containing an effective amount of therapeutic cationic peptide and directions for use of the kit.
19. A method of preparing a unimolecular nanoparticle comprising dispersing therapeutic cationic peptide within the unimolecular nanoparticle of claim 1 .
20. A method of treating a disease or condition in which PKM2 is overexpressed comprising administering an effective amount of a unimolecular nanoparticle of claim 11 .