Phase changing formulations of nucleic acid payloads
The present invention is based, at least in part, upon discovery of a process for identifying phase changing peptides. Such phase changing peptides are capable of enhancing in vitro and in vivo delivery of oligonucleotides (e.g., dsRNAs) in lipidic, vesicular, micellar and/or naked oligonucleotide formulations.
1. A method of disrupting a formulation or particle comprising:
preparing a formulation or particle comprising a cyclic charge-trapped peptide that is capable of linearizing when introduced into a reducing or low pH environment;
introducing said formulation or particle to a sufficiently reductive or low pH environment to linearize said cyclic peptide, wherein said linearization of said cyclic peptide disrupts said formulation or particle, thereby disrupting said formulation or particle;
wherein said cyclic peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-50.
2. The method of claim 1 , wherein said cyclic peptide is conjugated to an oligonucleotide.
3. The method of claim 1 , wherein said formulation or particle is a lipid formulation.
4. The method of claim 1 , wherein said formulation or particle is selected from the group consisting of a vesicle based formulation and a micelle based formulation.
5. The method of claim 1 , wherein said linearization of said cyclic peptide exposes a membrane integration domain within said peptide.
6. The method of claim 1 , wherein said linearization of said cyclic peptide exposes a pore-forming peptide.
7. The method of claim 1 , wherein said formulation or particle further comprises a dsRNA.
8. The method of claim 7 , wherein said dsRNA is conjugated to said cyclic peptide.
9. The method of claim 1 , wherein said formulation or particle further comprises a DsiRNA.
10. The method of claim 9 , wherein said DsiRNA is conjugated to said cyclic peptide.
11. The method of claim 7 , wherein said dsRNA is a dsRNA of 19-25 nucleotides.
12. The method of claim 9 , wherein said DsiRNA is a dsiRNA of 25-30 nucleotides.
13. The method of claim 2 , wherein said conjugated cyclic peptide is in an aqueous phase of the formulation.
14. The method of claim 2 , wherein said conjugated cyclic peptide is in a lipid or polymer phase of the formulation.
15. The method of claim 2 , wherein said dsiRNA and said conjugated cyclic peptide are in an aqueous phase of the formulation.
16. The method of claim 1 , wherein said cyclic peptide comprises a linkage selected from the group consisting of a disulfide linkage and a Citrulline-Valine linkage, wherein disruption of said linkage cyclizes said peptide.
17. The method of claim 1 , wherein said reducing or low pH environment constitutes an endocytic pathway compartment.
18. The method of claim 17 , wherein the endocytic pathway compartment is selected from the group consisting of a clathrin-coated vesicle (CCV), caveolae, a macropinocytic or phagocytic cell membrane invagination, a phagosome, an early endosome, a multivesicular body (MVB), an endosomal carrier vesicle (ECV), a late endosome and a lysosome.
19. The method of claim 1 , wherein said formulation optionally further comprises PEG.
20. The method of claim 1 , wherein said cyclic peptide is attached to PEG.