Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
Int J Pharm. 2022 Aug 25;624:121983. doi: 10.1016/j.ijpharm.2022.121983. Epub 2022 Jul 6.
Highly efficient and safe non-viral vectors for nucleic acids delivery have attracted much attention due to their potential applications in gene therapy, gene editing and vaccination against infectious diseases, and various materials have been investigated and designed as delivery vectors. Herein, we designed a series of branched amphiphilic peptides (BAPs) and tested their applications as pDNA/mRNA delivery vectors. The BAP structure was inspired by the phospholipids, in which lysine oligomers were used as the "polar head", segments containing phenylalanine, histidine and leucine were used as the "hydrophobic tails", and a lysine residue was used as the branching point. By comparing the gel retardation, particle sizes and zeta potentials of the BAP/pDNA complexes of the short-branch BAPs (BAP-V1 ∼ BAP-V4), we determined the optimal lysine oligomer was K. However, their cell transfection efficiencies were not satisfactory, and thus three long-branch BAPs (BAP-V5 ∼ BAP-V7) were further designed. In these long-branch BAPs, more hydrophobic residues were added and the overall amphiphilicity increased accordingly. The results showed that these three BAPs could effectively compact the nucleic acids, including both pDNA and mRNA, and all could transfect nucleic acids into HEK 293 cells, with low cytotoxicity. Among the three long-branch BAPs, BAP-V7 (bis(FFLFFHHH)-K-K) showed the best transfection efficiency at N/P = 10, which was better than the commercial transfection reagent PEI-25 K. These results indicate that increased amphiphilicity would also benefit for BAP mediated nucleic acid delivery. The designed BAPs provide more documents of such novel type of nucleic acids delivery vectors, which is worth of further investigation as a new gene theranostic platforms.
高效且安全的非病毒载体由于其在基因治疗、基因编辑和传染病疫苗接种等方面的潜在应用而受到广泛关注，各种材料已被研究和设计为递送载体。在此，我们设计了一系列支链两亲肽（BAP）并测试了它们作为 pDNA/mRNA 递送载体的应用。BAP 结构的灵感来自于磷脂，其中赖氨酸寡聚物被用作“极性头”，含有苯丙氨酸、组氨酸和亮氨酸的片段被用作“疏水尾巴”，而赖氨酸残基被用作支化点。通过比较短支链 BAP（BAP-V1～BAP-V4）的 BAP/pDNA 复合物的凝胶阻滞、粒径和 zeta 电位，我们确定了最佳的赖氨酸寡聚物为 K。然而，它们的细胞转染效率并不令人满意，因此进一步设计了三个长支链 BAP（BAP-V5～BAP-V7）。在这些长支链 BAP 中，添加了更多的疏水性残基，整体两亲性相应增加。结果表明，这三种 BAP 可以有效地压缩核酸，包括 pDNA 和 mRNA，并且都可以将核酸转染到 HEK 293 细胞中，具有低细胞毒性。在这三个长支链 BAP 中，BAP-V7（双（FFLFFHHH）-K-K）在 N/P=10 时表现出最佳的转染效率，优于商业转染试剂 PEI-25K。这些结果表明增加两亲性也有利于 BAP 介导的核酸递送。所设计的 BAP 为这种新型核酸递送载体提供了更多的文献资料，作为一种新的基因治疗平台值得进一步研究。
