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Stabilization of poly-L-lysine-based cancer-specific vectors via disulfide bond formation Kai LI1, Guoxi ZHAO1 ,Takeshi MORI1,2,3, Akihiro Kishimura 1,2,3,Yoshiki KATAYAMA1,2,3, 1 Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 2The Center for Future Chemistry, Kyushu University, 3International Research Center for Molecular Systems, Kyushu University Introduction We have proposed a novel gene targeting strategy, called “drug or gene delivery system responding to cellular signals (D-RECS)”. The systems can activate transgene in response to malfunctions of signal transduction occurring inside diseased cells. In the precious study we succeed in activating gene expression in response to several protein kinase1). However, the polyplex formed from polymeric carrier and plasmid DNA unfavorably dissociates in blood circulation. A promising approach to stabilize the polyplex will be reversibly cross-linking of the polyplex with disulfide bonds. Here a new gene carrier, poly-l-lysine (PLL) is modified with substrate peptide that can be phosphorylated by PKC and is also modified with 15-Carboxy-1-pentadecanethiol (15-CPDT) that can form disulfide bond to increase the stability of polyplex in blood circulation. Experiment PLL (Mw.15000-30000) was simply modified by peptide and 15-CPDT using HOBt and HBTU. After that, the protection group was removed by TFA. Four kinds of polymers with constant peptide modification ratio (5 mol%) and different 15-CPDT modification ratios (0, 5, 10 and 20 mol%) were prepred. The stability of complex that polymer and plasmid DNA formed (N/P=10) was evaluated by gel retardation analysis. Result and Discussion The release of pDNA from the by oxidation of disulfide bond was examined. Figure 1 shows the gel retardation assay of polyplexes fromed from each polymer. Free pDNA become weaker with the increase of 15-CDPT contents. On the contrary, the free pDNA bands were observed for reductive condition containing 20 mM DTT (lane 7-9). These results indicated that the disulfide cross-linking can stabilize the polyplexes and the polyplex can be weakened at the reductive condition 1) Riki Toita, et al, J. Am. Chem. Soc., 134, 1540-15417(2012)