Oligobenzylethylenimine enriches linear polyethylenimine with a pH-sensitive membrane-disruptive property and leads to enhanced gene delivery activity.

We report here the synthesis of a diblock linear polymer of oligo(benzylethylenimine)-b-polyethylenimine (OBzEI-PEI) and investigate its gene delivery properties. The linear copolymer OBzEI-PEI was prepared in a straightforward manner by acidic hydrolysis of a diblock polyoxazoline, which had been made by sequential polymerization of 4-benzyl-2-ethyl-2-oxazoline followed by 2-ethyl-2-oxazoline. pH titration and DNA complexation profiles of the new polymer are similar to regular linear PEIs, but with higher gene transfection efficiencies in various cell lines despite a decreased cellular uptake of plasmid DNA. Further experiments suggest that the OBzEI tail complements the intrinsic proton-sponge endosomolytic activities of PEI with an acid pH-sensitive membrane-perturbing activity.

Intracellular protein delivery with a dimerizable amphiphile for improved complex stability and prolonged protein release in the cytoplasm of adherent cell lines.

Direct delivery of functionally active proteins into cells represents an emerging strategy for laboratory investigation and therapeutic applications. For this purpose, we developed a novel amphiphile (CholCSper) consisting of cholesterol linked to carboxy-spermine by a cysteine. This amphiphile is dimerizable upon mild oxidation of the thiol to disulfide and it was used in formulation with DOPE to prepare an intracellular protein delivery system. The stabilization of the CholCSper assemblies by chemical conversion of CholCSper into its gemini amphiphile afforded the production of homogeneous assemblies with proteins whose sizes are easier to control. Furthermore, the cholesterol moiety has an effect on the density of the complexes formed with proteins and leads to a prolonged protein release in the cytoplasm of cells exposed to the protein carrier assemblies.