Chemo-Enzymatic Synthesis of Linear and Branched Cationic Peptides: Evaluation as Gene Carriers

Cationic peptides such as poly(l‐lysine) and poly(l‐arginine) are important tools for gene delivery since they can efficiently condense DNA. It is difficult to produce cationic peptides by recombinant bacterial expression, and its chemical synthesis requires several steps of protection/deprotection and toxic agents. Chemo‐enzymatic synthesis of peptides is a clean chemistry technique that allows fast production under mild conditions. With the aim to simplify the production of cationic peptides, the present work develops an enzymatic reaction which enables the synthesis of linear cationic peptides and, through terminal functionalization with tris(2‐aminoethyl)amine, of branched cationic peptide conjugates, which show improved DNA complex formation. Cytotoxicity and transfection efficiency of all the chemo‐enzymatically synthesized cationic peptides are evaluated for their novel use as gene delivery agents. Synthesized peptides exhibit transfection efficiencies comparable to previously reported monodisperse peptides. Chemo‐enzymatic synthesis opens the door for efficient production of cationic peptides for their use as gene delivery carriers. Branched and linear cationic peptides composed of l‐lysine and l‐arginine are synthesized by one‐pot chemo‐enzymatic reaction without using organic solvents or deprotection steps. Those peptides with wide distribution of molecular weights function as gene carriers and demonstrate similar transfection efficiencies to mono‐disperse cationic peptides.