Non-viral transfection reagents are continuously being developed in attempt to replace viral vectors. Among those non-viral vectors, dendrimers have gained increasing interest due to their unique molecular structure and multivalency. However, more improvements are still needed to achieve higher efficacy and lower toxicity. In this study, we have examined 18 peptide dendrimers conjugated to lipophilic moieties, such as fatty acids or hydrophobic amino acids, that were previously explored for siRNA. Reporter cells were employed to investigate the transfection of single strand splice-switching oligonucleotides (ONs) using these peptide dendrimers. Luciferase level changes reflecting efficiency varied with amino acid composition, stereochemistry, and complexation media used. 3rd generation peptide dendrimers with D-amino acid configuration were superior to L-form. Lead formulations with 3rd generation, D-amino acid peptide dendrimers increased the correction level of the delivered ON up to 93-fold over untreated HeLa Luc/705 cells with minimal toxicity. To stabilize the formed complexes, Polyvinyl alcohol 18 (PVA18) polymer was added. Although PVA18 addition increased activity, toxicity when using our best candidates G 2,3KL-(Leu)₄ (D) and G 2,3KL-diPalmitamide (D) was observed. Our findings demonstrate the potential of lipid-conjugated, D-amino acid-containing peptide dendrimers to be utilized as an effective and safe delivery vector for splice-switching ONs.

中文翻译：

---

亲脂性肽树状聚合物的剪接转换寡核苷酸的交付。

非病毒转染试剂正在不断开发，以试图替代病毒载体。在那些非病毒载体中，树枝状大分子由于其独特的分子结构和多价而越来越受到关注。然而，仍需要更多的改进以实现更高的功效和更低的毒性。在这项研究中，我们检查了18种与树枝状亲脂性分子（例如脂肪酸或疏水性氨基酸）缀合的肽树状大分子，这些多肽树状大分子先前已被研究用于siRNA。报告细胞被用于研究使用这些肽树状聚合物的单链剪接转换寡核苷酸（ON）的转染。萤光素酶水平的变化反映了效率随所用氨基酸组成，立体化学和络合介质的变化而变化。具有D-氨基酸构型的第三代肽树状聚合物优于L-形式。与未处理的HeLa Luc / 705细胞相比，具有第三代D-氨基酸肽树状聚合物的先导配方将传递的ON的校正水平提高了93倍，毒性最小。为了稳定形成的络合物，添加了聚乙烯醇18（PVA18）聚合物。尽管添加PVA18可以提高活性，但使用我们最好的候选药物G 2,3KL-（Leu）时的毒性观察到图₄（D）和G 2,3KL-二棕榈酰胺（D）。我们的发现证明了脂质缀合的，包含D-氨基酸的肽树状聚合物的潜力，可以用作剪接转换ON的有效和安全的载体。