Perche Federico, Biswas Swati, Patel Niravkumar R, Torchilin Vladimir P
Center for Pharmaceutical Biotechnology and Nanomedicine, School of Pharmacy, Bouve College of Health Sciences, Northeastern University, Boston, MA, USA.
Department of Pharmaceutical Sciences, School of Pharmacy, Bouve College of Health Sciences, Northeastern University, Boston, MA, USA.
Methods Mol Biol. 2016;1372:139-62. doi: 10.1007/978-1-4939-3148-4_12.
A wide variety of nanomedicine has been designed for cancer therapy. Herein, we describe the synthesis and evaluation of a hypoxia-responsive copolymer for siRNA delivery (Perche et al., Angew Chem Int Ed Engl 53:3362-3366, 2014). The synthesis is achieved using established coupling chemistry and accessible purification procedures. A polyelectrolyte-lipid conjugate (polyethyleneimine 1.8 kDa-dioleyl-phosphatidylinositol, PEI-PE) and polyethylene glycol 2000 (PEG) were assembled via the hypoxia-sensitive azobenzene (Azo) unit to obtain the PEG-Azo-PEI-DOPE copolymer. This copolymer can condense siRNA and shows hypoxia-induced cellular internalization and reporter gene downregulation in vitro and tumor accumulation in vivo after parenteral administration (Perche et al., Angew Chem Int Ed Engl 53:3362-3366, 2014). We also detail procedures to evaluate hypoxia-targeted polymers both in monolayer cultures, cancer cell spheroids and in tumor xenografts murine models.
人们已设计出各种各样的纳米药物用于癌症治疗。在此，我们描述了一种用于递送小干扰RNA（siRNA）的缺氧响应性共聚物的合成与评估（佩尔什等人，《德国应用化学》国际版 英文 53:3362 - 3366，2014年）。合成过程采用成熟的偶联化学方法和易于操作的纯化程序来实现。通过缺氧敏感的偶氮苯（Azo）单元组装聚电解质 - 脂质共轭物（1.8 kDa聚乙烯亚胺 - 二油酰磷脂酰肌醇，PEI - PE）和聚乙二醇2000（PEG），以获得PEG - Azo - PEI - DOPE共聚物。这种共聚物能够凝聚siRNA，并在体外显示出缺氧诱导的细胞内化以及报告基因下调，在肠胃外给药后在体内显示出肿瘤蓄积（佩尔什等人，《德国应用化学》国际版 英文 53:3362 - 3366，2014年）。我们还详细介绍了在单层培养、癌细胞球体以及肿瘤异种移植小鼠模型中评估缺氧靶向聚合物的程序。
