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.

用于杜氏肌营养不良症的反义寡核苷酸（AON）治疗在临床前和临床试验中引起了极大的关注，但由于递送效率低下，其治疗应用仍然受到限制。在这项研究中，我们调查了一些皂苷在体外和体内改善反义磷酸二酰胺吗啉代寡聚物（PMO）传递性能的潜力。结果表明，这些皂角苷，特别是洋地黄皂苷和番茄碱，与Endoporter介导的PMO体外递送相比，提高了PMO的递送效率。在mdx中进一步观察到PMO靶向肌营养不良蛋白外显子23递送的显着增强与单独的PMO相比，使用Digitonin的小鼠最多可提高7倍。洋地黄皂苷和甘草甜素的细胞毒性在体外低于Endoporter ，并且在测试浓度下体内并未明确检测到。这些结果表明，皂苷在分子大小和组成上的优化是实现增强的PMO外显子跳跃效率的关键因素。更高的效率和更低的毒性赋予了皂苷作为基因/ AON传递增强剂的功效，可用于治疗肌营养不良或其他疾病。