Bhattarai Prashant, Vance Dylan, Hatefi Arash, Khaw Ban An
a Department of Pharmaceutical Sciences , Northeastern University , Boston , MA , USA.
b Department of Biology , College of Sciences, Northeastern University , Boston , MA , USA.
J Drug Target. 2017 Jun;25(5):436-450. doi: 10.1080/1061186X.2016.1271421. Epub 2017 Jan 5.
Drug resistance is a common phenomenon that occurs in cancer chemotherapy. Delivery of chemotherapeutic agents as polymer pro-drug conjugates (PPDCs) pretargeted with bispecific antibodies could circumvent drug resistance in cancer cells. To demonstrate this approach to overcome drug resistance, Paclitaxel (Ptxl)-resistant SKOV3 TR human ovarian- and doxorubicin (Dox)-resistant MCF7 ADR human mammary-carcinoma cell lines were used. Pre-targeting over-expressed biotin or HER2/neu receptors on cancer cells was conducted by biotinylated anti-DTPA or anti-HER2/neu affibody - anti-DTPA Fab bispecific antibody complexes. The targeting PPDCs are either D-Dox-PGA or D-Ptxl-PGA. Cytotoxicity studies demonstrate that the pretargeted approach increases cytotoxicity of Ptxl or Dox in SKOV3 TR or MCF7 ADR resistant cell lines by 5.4 and 27 times, respectively. Epifluorescent microscopy - used to track internalization of D-Dox-PGA and Dox in MCF7 ADR cells - shows that the pretargeted delivery of D-Dox-PGA resulted in a 2- to 4-fold increase in intracellular Dox concentration relative to treatment with free Dox. The mechanism of internalization of PPDCs is consistent with endocytosis. Enhanced drug delivery and intracellular retention following pretargeted delivery of PPDCs resulted in greater tumor cell toxicity in the current in vitro studies.
耐药性是癌症化疗中常见的现象。将化疗药物作为双特异性抗体预靶向的聚合物前药偶联物(PPDCs)递送,可以规避癌细胞中的耐药性。为了证明这种克服耐药性的方法,使用了对紫杉醇(Ptxl)耐药的SKOV3 TR人卵巢癌细胞系和对阿霉素(Dox)耐药的MCF7 ADR人乳腺癌细胞系。通过生物素化的抗二乙三胺五乙酸(DTPA)或抗HER2/neu亲和体-抗DTPA Fab双特异性抗体复合物对癌细胞上过度表达的生物素或HER2/neu受体进行预靶向作用。靶向PPDCs为D-Dox-PGA或D-Ptxl-PGA。细胞毒性研究表明,预靶向方法使Ptxl或Dox在SKOV3 TR或MCF7 ADR耐药细胞系中的细胞毒性分别提高了5.4倍和27倍。用于追踪D-Dox-PGA和Dox在MCF7 ADR细胞中内化情况的落射荧光显微镜显示,相对于游离Dox处理,D-Dox-PGA的预靶向递送使细胞内Dox浓度提高了2至4倍。PPDCs的内化机制与内吞作用一致。在当前的体外研究中,PPDCs预靶向递送后增强的药物递送和细胞内滞留导致了更大的肿瘤细胞毒性。