Saponins as Natural Adjuvant for Antisense Morpholino Oligonucleotides Delivery in vitro and in mdx Mice

Antisense oligonucleotide (AON) therapy for Duchenne muscular dystrophy has drawn great attention in preclinical and clinical trials, but its therapeutic applications are still limited due to inefficient delivery. In this study, we investigated a few Saponins for their potential to improve delivery performance of an antisense phosphorodiamidate morpholino oligomer (PMO) both in vitro and in vivo. The results showed that these Saponins, especially Digitonin and Tomatine, improve the delivery efficiency of PMO comparable to Endoporter-mediated PMO delivery in vitro. The significant enhancement of PMO targeting to dystrophin exon 23 delivery was further observed in mdx mice up to 7-fold with the Digitonin as compared to PMO alone. Cytotoxicity of the Digitonin and Glycyrrhizin was lower than Endoporter in vitro and not clearly detected in vivo under the tested concentrations. These results demonstrate that optimization of Saponins in molecular size and composition are key factors to achieve enhanced PMO exon-skipping efficiency. The higher efficiency and lower toxicity endow Saponins as gene/AON delivery enhancing agents for treating muscular dystrophy or other diseases.