Manickam Devika Soundara, Oupický David
Department of Pharmaceutical Sciences, Wayne State University, Detroit, Michigan 48202, USA.
Bioconjug Chem. 2006 Nov-Dec;17(6):1395-403. doi: 10.1021/bc060104k.
Polyplexes sensitive to redox potential gradients represent promising gene delivery vectors. High molecular weight polypeptides containing disulfide bonds in the backbone were synthesized by an oxidative copolymerization of a histidine-rich peptide (HRP) and a nuclear localization sequence (NLS) peptide. The synthetic approach allowed an easy synthesis of reducible copolypeptides (rCPP) with different relative contents of the HRP and NLS sequences. Cytotoxicity and transfection activity of rCPP-based DNA polyplexes were evaluated in vitro. In comparison with control polyethylenimine (PEI), only minimum toxic effects of rCPPs were observed on the metabolic activity and membrane integrity of human endothelial cells. These findings are predominantly ascribed to favorable structural features like lower charge density and higher chain rigidity of the rCPPs compared to PEI and also to a reductive intracellular and plasma membrane degradation. Transfection activity in all tested cell lines increased with increasing content of the HRP sequence in the rCPPs, while no clearly measurable effect of the NLS sequence was found.
对氧化还原电位梯度敏感的多聚体是很有前景的基因传递载体。通过富含组氨酸的肽(HRP)与核定位序列(NLS)肽的氧化共聚反应,合成了主链中含有二硫键的高分子量多肽。该合成方法能够轻松合成具有不同HRP和NLS序列相对含量的可还原共多肽(rCPP)。在体外评估了基于rCPP的DNA多聚体的细胞毒性和转染活性。与对照聚乙烯亚胺(PEI)相比,仅观察到rCPP对人内皮细胞的代谢活性和膜完整性有最小的毒性作用。这些发现主要归因于与PEI相比,rCPP具有较低的电荷密度和较高的链刚性等有利的结构特征,以及细胞内和质膜的还原性降解。在所有测试细胞系中,转染活性随着rCPP中HRP序列含量的增加而增加,而未发现NLS序列有明显可测量的影响。