Efficient intracellular delivery of antisense oligonucleotide and pharmaceutical factors

点击翻译按钮获取中文摘要

As a carrier for intracellular delivery of an antisense and plasmid DNA,synthetic oligopeptides were examined. The oligopeptide which consisted of nine amino acids was synthesized as the carrier with a peptide synthesizer. Various peptide which consisted of lysine, leucine, cysteine, and tryptophane were synthesized and purified by the liquid chromatoqraphy which the reverse-aspect column was installed. The aptitude as the intracellular introduction carrier of the deoxyribonucleic acid with this synthetic peptide was examined by using the HeLaS3 cell by the plasmid DNA which contained the CAT gene.The gene expression was not observed in peptide which consisted of lysine and leucine, but observed in peptide contained cysteine. The dimer was found with the mass spectrum and the gene expression ability disappeared when the disulfide linkage of the peptide was cut by the DTT processing. The CD measurement showed that peptide took the structure which consisted of two peptide chains. As a result of the gel electrophoresis, the DNA and peptide formed complex. It were confirmed the complex was taken into intracellular from the observation with confocus laser microscope. The gene expression have been observed even by cells another of the HeLaS3, such as CHO,L929, COS7 and HepG2 cells. The introduction efficiency has improved when tryptophane is introduced into peptide. It has been understood that the oligopeptide which consists of the basic amino acid and the hydrophobic amino acid with the disulfide bond has the ability to introduce the deoxyribonucleic acid into intracellular. It will be studied on reletionship between the secondary structure of peptide and the interaction with the deoxyribonucleic acid further in the future.