Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrhenius väg 21A, SE-10691 Stockholm, Sweden.
Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrhenius väg 21A, SE-10691 Stockholm, Sweden; Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayudhya Road, 10400 Bangkok, Thailand.
Int J Pharm. 2014 Apr 10;464(1-2):111-6. doi: 10.1016/j.ijpharm.2014.01.018. Epub 2014 Jan 22.
A series of novel, amphipathic cell-penetrating peptides was developed based on a combination of the model amphipathic peptide sequence and modifications based on the strategies developed for PepFect and NickFect peptides. The aim was to study the role of amphipathicity for peptide uptake and to investigate if the modifications developed for PepFect peptides could be used to improve the uptake of another class of cell-penetrating peptides. The peptides were synthesized by solid phase peptide synthesis and characterized by circular dichroism spectroscopy. Non-covalent peptide-plasmid complexes were formed by co-incubation of the peptides and plasmids in water solution. The complexes were characterized by dynamic light scattering and cellular uptake of the complexes was studied in a luciferase-based plasmid transfection assay. A quantitative structure-activity relationship (QSAR) model of cellular uptake was developed using descriptors including hydrogen bonding, peptide charge and positions of nitrogen atoms. The peptides were found to be non-toxic and could efficiently transfect cells with plasmid DNA. Cellular uptake data was correlated to QSAR predictions and the predicted biological effects obtained from the model correlated well with experimental data. The QSAR model could improve the understanding of structural requirements for cell penetration, or could potentially be used to predict more efficient cell-penetrating peptides.
基于模型两亲肽序列和基于 PepFect 和 NickFect 肽开发策略的修饰组合，开发了一系列新型两亲性细胞穿透肽。目的是研究肽摄取的两亲性作用，并研究为 PepFect 肽开发的修饰是否可用于改善另一类细胞穿透肽的摄取。肽通过固相肽合成合成，并通过圆二色性光谱进行表征。通过在水溶液中共孵育肽和质粒来形成非共价肽-质粒复合物。通过动态光散射对复合物进行表征，并在基于荧光素酶的质粒转染测定中研究复合物的细胞摄取。使用包括氢键、肽电荷和氮原子位置在内的描述符，开发了细胞摄取的定量构效关系（QSAR）模型。肽被发现是无毒的，并且可以有效地用质粒 DNA 转染细胞。将细胞摄取数据与 QSAR 预测相关联，并且从模型获得的预测生物效应与实验数据很好地相关。QSAR 模型可以提高对细胞穿透结构要求的理解，或者可能用于预测更有效的细胞穿透肽。
