Jin Yao, Adams Friederike, Nguyen Anny, Sturm Sebastian, Carnerio Simone, Müller-Caspary Knut, Merkel Olivia M
Department of Pharmacy, Ludwig-Maximilians-University Munich, Pharmaceutical Technology and Biopharmaceutics Butenandtstr. 5-13 81377 Munich Germany
Department of Chemistry and Centre for NanoScience, Ludwig-Maximilians-University Munich Butenandtstr. 11 81377 Munich Germany.
Nanoscale Adv. 2023 Sep 7;5(19):5256-5262. doi: 10.1039/d3na00272a. eCollection 2023 Sep 26.
Small interfering RNA (siRNA) can trigger RNA interference (RNAi) to therapeutically silence disease-related genes in human cells. The approval of siRNA therapeutics by the FDA in recent years generated a new hope in novel and efficient siRNA therapeutics. However, their therapeutic application is still limited by the lack of safe and efficient transfection vehicles. In this study, we successfully synthesized a novel amphiphilic poly(β-amino ester) based on the polyamine spermine, hydrophobic decylamine and 1,4-butanediol diacrylate, which was characterized by H NMR spectroscopy and size exclusion chromatography (SEC, = 6000 Da). The polymer encapsulated siRNA quantitatively from N/P 5 on as assessed by fluorescence intercalation while maintaining optimal polyplex sizes and zeta potentials. Biocompatibility and cellular delivery efficacy were also higher than those of the commonly used cationic, hyperbranched polymer polyethylenimine (PEI, 25 kDa). Optimized formulations mediated around 90% gene silencing in enhanced green fluorescence protein expressing H1299 cells (H1299-eGFP) as determined by flow cytometry. These results suggest that spermine-based, amphiphilic poly(β-amino ester)s are very promising candidates for efficient siRNA delivery.
小干扰RNA（siRNA）可触发RNA干扰（RNAi），从而在人类细胞中使疾病相关基因实现治疗性沉默。近年来，FDA对siRNA疗法的批准为新型高效的siRNA疗法带来了新希望。然而，其治疗应用仍受到缺乏安全高效转染载体的限制。在本研究中，我们成功合成了一种新型两亲性聚（β-氨基酯），该聚合物基于多胺精胺、疏水性癸胺和1,4-丁二醇二丙烯酸酯合成，通过核磁共振氢谱（1H NMR）和尺寸排阻色谱（SEC，Mw = 6000 Da）对其进行了表征。通过荧光嵌入法评估，该聚合物在N/P为5时即可定量包封siRNA，同时保持最佳的纳米颗粒尺寸和zeta电位。其生物相容性和细胞递送效率也高于常用的阳离子超支化聚合物聚乙烯亚胺（PEI，25 kDa）。通过流式细胞术测定，优化后的制剂在表达增强型绿色荧光蛋白的H1299细胞（H1299-eGFP）中可介导约90%的基因沉默。这些结果表明，基于精胺的两亲性聚（β-氨基酯）是高效递送siRNA的极具潜力的候选材料。
