Lv Hui, Zhu Qing, Liu Kewu, Zhu Manman, Zhao Wenfang, Mao Yuan, Liu Kehai
Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai, People's Republic of China.
Heilongjiang Forest By-Product and Speciality Institute, Mudanjiang, People's Republic of China.
Int J Nanomedicine. 2014 Mar 11;9:1311-22. doi: 10.2147/IJN.S59726. eCollection 2014.
A degradable polyethylenimine (PEI) derivative coupled to a bifunctional peptide R13 was developed to solve the transfection efficiency versus cytotoxicity and tumor-targeting problems of PEI when used as a gene vector.
We crossed-linked low molecular weight PEI with N-octyl-N-quaternary chitosan (OTMCS) to synthesize a degradable PEI derivative (OTMCS-PEI), and then used a bifunctional peptide, RGDC-TAT (49-57) called R13 to modify OTMCS-PEI so as to prepare a new gene vector, OTMCS-PEI-R13. This new gene vector was characterized by various physicochemical methods. Its cytotoxicity and gene transfection efficiency were also determined both in vitro and in vivo.
The vector showed controlled degradation and excellent buffering capacity. The particle size of the OTMCS-PEI-R13/DNA complexes was around 150-250 nm and the zeta potential ranged from 10 mV to 30 mV. The polymer could protect plasmid DNA from being digested by DNase I at a concentration of 23.5 U DNase I/μg DNA. Further, the polymer was resistant to dissociation induced by 50% fetal bovine serum and 400 μg/mL sodium heparin. Compared with PEI 25 kDa, the OTMCS-PEI-R13/DNA complexes showed higher transfection efficiency both in vitro and in vivo. Further, compared with OTMCS-PEI, distribution of OTMCS-PEI-R13 at tumor sites was markedly enhanced, indicating the tumor-targeting specificity of R13.
OTMCS-PEI-R13 could be a potential candidate as a safe and efficient gene delivery carrier for gene therapy.
开发了一种与双功能肽R13偶联的可降解聚乙烯亚胺（PEI）衍生物，以解决PEI作为基因载体时的转染效率与细胞毒性以及肿瘤靶向问题。
我们将低分子量PEI与N-辛基-N-季铵化壳聚糖（OTMCS）交联以合成可降解的PEI衍生物（OTMCS-PEI），然后使用一种名为R13的双功能肽RGDC-TAT（49-57）修饰OTMCS-PEI，从而制备一种新的基因载体OTMCS-PEI-R13。通过各种物理化学方法对这种新的基因载体进行了表征。还在体外和体内测定了其细胞毒性和基因转染效率。
该载体显示出可控的降解性和出色的缓冲能力。OTMCS-PEI-R13/DNA复合物的粒径约为150-250nm，ζ电位范围为10mV至30mV。该聚合物能够在23.5U DNase I/μg DNA的浓度下保护质粒DNA不被DNase I消化。此外，该聚合物对50%胎牛血清和400μg/mL肝素钠诱导的解离具有抗性。与25kDa的PEI相比，OTMCS-PEI-R13/DNA复合物在体外和体内均表现出更高的转染效率。此外，与OTMCS-PEI相比，OTMCS-PEI-R13在肿瘤部位的分布明显增强，表明R13具有肿瘤靶向特异性。
OTMCS-PEI-R13可能是一种潜在的候选物，可作为基因治疗中安全有效的基因递送载体。
