Rennert Robert, Neundorf Ines, Jahnke Heinz-Georg, Suchowerskyj Philipp, Dournaud Pascal, Robitzki Andrea, Beck-Sickinger Annette G
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstr. 34, 04103 Leipzig, Germany.
ChemMedChem. 2008 Feb;3(2):241-53. doi: 10.1002/cmdc.200700216.
Now that the human genome has been decoded, the demand for novel therapeutic concepts, such as gene and stem cell therapy, is higher than ever before. Although new and better pharmaceutical agents are available, their efficient delivery to the intracellular site of action is still a serious challenge. A possible solution to this problem is the use of cell-penetrating peptides as delivery vectors, including derivatives of human calcitonin (hCT). The aim of this study was to synthesise novel branched hCT-derived peptides for the noncovalent delivery of nucleic acids. The uptake of the resulting oligocationic peptides into various cell lines as well as primary cells was monitored by fluorescence microscopy. To determine the appropriate peptide-plasmid charge ratios for efficient cell transfection, electromobility shift assays were carried out. Finally, flow cytometric and fluorescence microscopic studies of gene expression highlighted two novel hCT-derived peptides as highly effective in the delivery of noncovalently complexed plasmid DNA. Thus, the absence of cytotoxicity paired with highly efficient cell internalisation and transfection rates, in primary cells as well, make both peptides powerful candidates as drug delivery vectors, especially for plasmid DNA, for both in vivo and ex vivo therapeutic applications.
既然人类基因组已被解码，对诸如基因疗法和干细胞疗法等新型治疗理念的需求比以往任何时候都更高。尽管有了新的更好的药物制剂，但将它们有效地递送至细胞内作用位点仍然是一个严峻的挑战。解决这个问题的一个可能办法是使用细胞穿透肽作为递送载体，包括人降钙素（hCT）的衍生物。本研究的目的是合成新型分支hCT衍生肽用于核酸的非共价递送。通过荧光显微镜监测所得寡阳离子肽进入各种细胞系以及原代细胞的情况。为了确定有效细胞转染的合适肽-质粒电荷比，进行了电泳迁移率变动分析。最后，对基因表达的流式细胞术和荧光显微镜研究突出显示了两种新型hCT衍生肽在非共价复合质粒DNA递送方面非常有效。因此，既没有细胞毒性，又在原代细胞中具有高效的细胞内化和转染率，这使得这两种肽成为强大的药物递送载体候选物，特别是用于体内和体外治疗应用的质粒DNA。
