Synthesis and evaluation of new phosphonolipid compounds for gene delivery

The preparation of a series of novel water soluble cationic lipid derivatives possessing phosphonate ester groups linked to the para-position of N-methyl pyridinium moieties and bearing either identical or different alkyl chains is reported. The obtained phospholipids were tested for transfection efficiency into three different mammalian cell lines alone and in conjunction with diphytanoylphosphatidylethanolamine (DiPPE) or dioleylphosphatidylethanolamine (DOPE), using an assay adapted for 96-well microplates based on the detection of a colorimetric change caused by the production of a chromogen induced by expressed secreted human placental alkaline phosphatase. In our conditions, the highest transfection activities of cells HEK293 and hard-to-transfect cell lines B16 and CHO were achieved with a 4-phosphonobutylpyridinium compound used at 1:5, 1:10 or 3:6 DNA/lipid ratio bearing two myristyl chains in the presence of the fusogenic helper lipid DiPPE.

Graphical abstractA series of cationic lipids containing phosphonate esters were prepared. Lipids were assessed as vectors for transfecting DNA to various cell lines, under different conditions, in a 96-well format.Figure optionsDownload full-size imageDownload as PowerPoint slide