Method for manufacturing linear polyethylenimine (pei) for transfection purpose and linear pei obtained with such method

Abstract

The invention concerns a method of synthesising and preparing linear polyethylenimine (PEI) for use as a transfection vector, and the product obtained with such a method. It comprises drying a monomer 2-ethyl-2-oxazoline and polymerising said monomer for obtaining poly (2- ethyl-2-oxazoline) (PEOX) by : using acetonitrile as solvent, adding a dried initiator of the reaction of polymerisation, and mixing them altogether, purifying said obtained PEOX by evaporation, while performing at least three times successive washing / precipitation steps with methanol and diethyl ether and corresponding filtrations, in order to obtain (i), by performing 1H-NMR tests, correct identification of said PEOX polymer, confirmation of absence of monomer to a level <1.0% and confirmation of absence of solvent to a level <5.0% and (ii), by performing Gel Permeation Chromatography, a mean of molecular weight (Mw) >23,000 Da and polydispersity (Mw / Mn) of said PEOX < 1.5, hydrolysing said PEOX.

Description

technical field [0001] The present invention relates to the manufacture and quality control of linear polyethyleneimine (PEI) for transfection applications. [0002] The invention also relates to products resulting from such manufacturing methods, more particularly for use in in vivo applications including but not limited to nucleic acid-based therapeutics. [0003] This application is a non-provisional application related to and claiming priority from earlier US provisional application US 60 / 952993. Background technique [0004] Polyethyleneimine (PEI) is an organic macromolecule with the potential for high cationic charge density. Every third atom of PEI is a protonatable amino nitrogen. PEI is capable of trapping DNA and, due to the close location of many linker amino groups, PEI retains a large buffering capacity at almost any pH. [0005] PEI alone is an efficient vector for delivering DNA plasmids in vitro and in vivo. [0006] PEI compacts DNA into positively char...

AI Technical Summary

Problems solved by technology

[0010] However, such PEIs tend to be inefficient when used for transfection purposes, i.e. less than 10% of the produced product is successfully transferred into cells

[0011] Especially high molecular weight PEI (i.e. >10,000Da) is less efficient

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