PHASE CHANGING FORMULATIONS OF NUCLEIC ACID PAYLOADS

摘要

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.

主权项

1. A method of identifying a phase changing peptide based on a peptide's efficacy in delivering an oligonucleotide to the cytoplasm of a cell in a subject after administration of a formulation comprising said oligonucleotide and said peptide, comprising the steps of:
(i) providing a test peptide, wherein said peptide is a charge-trapped peptide; (ii) preparing a formulation comprising said oligonucleotide and said test peptide; (iii) comparing the activity of said formulation with the activity of a control formulation which lacks said test peptide, in one or more of the following assays:
(a) an assay measuring/evaluating internalization of said oligonucleotide into said cell,(b) an assay measuring/evaluating leakiness of said cell induced by said formulation of step (ii),(c) an assay measuring/evaluating in vitro target knockdown,(d) an assay measuring/evaluating in vivo target knockdown,(e) an assay measuring/evaluating in vivo efficacy; and (iv) identifying the test peptide as a phase changing peptide if the formulation of step (ii) displays an increased activity in at least one of the assays (a)-(e) relative to the control formulation.