Optimizing pDNA Delivery by fine tunning Trehalose-Based Vector Structures and Formulations

In recent years, nucleic acid therapeutics have made a remarkable clinical impact, most notably exemplified by the mRNA-based vaccines developed against SARS-CoV-2. Much of this success is attributed to the development of suitable delivery carriers that help nucleic acids overcoming physiological barriers en route to their targets. Although viruses have traditionally served as a blueprint for delivery systems, non-viral platforms such as lipid nanoparticles (LNPs) are gaining prominence due to their greater potential for functional customization through synthetic and pharmaceutical technologies. Conformationally locked carbohydrates like cyclodextrins or α,α′-trehalose have emerged as versatile molecular scaffolds for designing such carriers.1,2 Their bifacial architecture enables the diversity-oriented construction of Janus amphiphiles that combine functional modules capable of (i) electrostatic interaction with nucleic acids and (ii) hydrophobic interactions to drive condensation. In this work, we present a novel family of α,α′-trehalose-based vectors bearing cationic heads and lipophilic tails at the secondary and primary position, respectively (Figure 1). Their pDNA delivery properties are strongly dependent on formulation conditions and correlate with key structural features of the amphiphiles, including the pKa of their cationic groups, and the density and length of their hydrophobic domains.