Nah Jae-Woon, Yu Lei, Han Sang-oh, Ahn Cheol-Hee, Kim Sung Wan
Department of Polymer Science and Engineering, Sunchon National University, 540-742, Chonnam, Republic of Korea.
J Control Release. 2002 Jan 17;78(1-3):273-84. doi: 10.1016/s0168-3659(01)00499-0.
Artery wall binding peptide (AWBP; Cys-Gly-Arg-Ala-Leu-Val-Asp-Thr-Leu-Lys-Phe-Val-Thr-Gln-Ala-Glu-Gly-Ala-Lys), a specific targeting peptide, was conjugated to poly(ethylene glycol)-grafted-poly(L-lysine) (PEG-g-PLL) to enhance the gene transfer to artery wall cells. AWBP-PEG-PLL was synthesized by the reaction between the vinylsulfone group of PEG-g-PLL and the thiol group of cysteine in AWBP. 1H-NMR analysis confirmed the composition of the obtained polymer and indicated that four mol of AWBP were reacted to one mole of VS-PEG-PLL. The particles of AWBP-PEG-PLL/pDNA complexes were determined spherical with a size of approximately 100 nm by dynamic light scattering (DLS) and atomic force microscopy (AFM). Agarose gel retardation assay indicated that AWBP-PEG-PLL was able to condense plasmid DNA and reach complete complexation at and above a charge ratio 1/1 (+/-). Transfection efficiency of AWBP-PEG-PLL/pDNA complexes was 150-180 times higher than that of control systems, such as PEG-g-PLL/pDNA and PLL/pDNA, in both bovine aorta endothelial cells and smooth muscle cells. Luciferase activities of AWBP-PEG-PLL depended on the amount of free AWBP, while those of the control carriers such as PLL and PEG-g-PLL were not affected by free AWBP. These results supported that gene transfer of AWBP-PEG-PLL/pDNA complexes to bovine aorta wall cells was mediated by specific artery wall cell receptor-mediated endocytosis.
动脉壁结合肽（AWBP；半胱氨酸-甘氨酸-精氨酸-丙氨酸-亮氨酸-缬氨酸-天冬氨酸-苏氨酸-亮氨酸-赖氨酸-苯丙氨酸-缬氨酸-苏氨酸-谷氨酰胺-丙氨酸-谷氨酸-甘氨酸-丙氨酸-赖氨酸），一种特异性靶向肽，与聚乙二醇接枝聚-L-赖氨酸（PEG-g-PLL）偶联，以增强向动脉壁细胞的基因转移。AWBP-PEG-PLL通过PEG-g-PLL的乙烯砜基团与AWBP中半胱氨酸的巯基之间的反应合成。1H-NMR分析证实了所得聚合物的组成，并表明四摩尔的AWBP与一摩尔的VS-PEG-PLL反应。通过动态光散射（DLS）和原子力显微镜（AFM）测定，AWBP-PEG-PLL/pDNA复合物颗粒呈球形，大小约为100nm。琼脂糖凝胶阻滞试验表明，AWBP-PEG-PLL能够凝聚质粒DNA，并在电荷比为1/1（+/-）及以上时达到完全络合。在牛主动脉内皮细胞和平滑肌细胞中，AWBP-PEG-PLL/pDNA复合物的转染效率比对照系统（如PEG-g-PLL/pDNA和PLL/pDNA）高150-180倍。AWBP-PEG-PLL的荧光素酶活性取决于游离AWBP的量，而对照载体如PLL和PEG-g-PLL的荧光素酶活性不受游离AWBP的影响。这些结果支持AWBP-PEG-PLL/pDNA复合物向牛主动脉壁细胞的基因转移是由特异性动脉壁细胞受体介导的内吞作用介导的。
