Xue Weihua, Diao Huajia, Chen Xi, Wang Chunming, Chen Jiangning, Zhang Junfeng
State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing, China.
Eur J Pharm Biopharm. 2007 Jun;66(3):327-33. doi: 10.1016/j.ejpb.2006.11.016. Epub 2006 Dec 1.
PSI, as the potential peptide-like intermediate, is subject to simple chemical modification in order to obtain good non-viral carriers for gene delivery. This paper describes the facile synthesis and preliminary evaluation of alpha,beta-poly (3-dimethylaminopropyl-D,L-aspartamide) (PDAI) as a vector. Reaction of PSI with 3-dimethylamino-1-propylamine afforded PDAI in N,N-dimethylformamide (DMF) solution. Such biophysical properties of PDAI/DNA complexes as the particle size and the zeta potential were determined by dynamic light scattering assay. The complexes prepared at weight ratios ranging from 2 to 3 have an average size of around 200 nm and a zeta potential of around 10.0 mV. Gel electrophoresis assays confirmed that PDAI could compact DNA to form the complexes and protect DNA from enzymatic degradation by DNase I at the weight ratio above 2.0. Furthermore, PDAI was found to transfect HepG2 cells at a much higher efficiency than commercially available polyethylenimine (PEI) (W(w)=75,000 Da). MTT cytotoxicity assay demonstrated that PDAI also showed much less toxicity than did PEI, suggesting that PDAI is a new class of transfection reagent to be used as a safe vector.
PSI作为潜在的肽样中间体,需要进行简单的化学修饰以获得用于基因递送的良好非病毒载体。本文描述了α,β-聚(3-二甲基氨基丙基-D,L-天冬酰胺)(PDAI)作为载体的简便合成方法及初步评估。PSI与3-二甲基氨基-1-丙胺在N,N-二甲基甲酰胺(DMF)溶液中反应生成PDAI。通过动态光散射测定法确定了PDAI/DNA复合物的粒径和zeta电位等生物物理性质。以2至3的重量比制备的复合物平均粒径约为200 nm,zeta电位约为10.0 mV。凝胶电泳分析证实,PDAI在重量比高于2.0时能够压缩DNA形成复合物,并保护DNA免受DNase I的酶促降解。此外,发现PDAI转染HepG2细胞的效率比市售聚乙烯亚胺(PEI)(W(w)=75,000 Da)高得多。MTT细胞毒性试验表明,PDAI的毒性也比PEI小得多,这表明PDAI是一种新型的转染试剂,可作为安全载体使用。