Tapia-Moreno Alejandro, Juarez-Moreno Karla, Gonzalez-Davis Oscar, Cadena-Nava Ruben D, Vazquez-Duhalt Rafael
Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Ensenada, Baja California, Mexico.
Biotechnol J. 2017 Jun;12(6). doi: 10.1002/biot.201600706. Epub 2017 May 9.
Most of the drugs used in chemotherapy should be activated by a transformation catalyzed by cytochrome P450 (CYP) enzymes. In this work, bacteriophage P22 virus-like particles (VLPs) containing CYP activity, immunologically inert and functionalized in order to be recognized by human cervix carcinoma cells and human breast adenocarcinoma cells were designed. The CYP was encapsulated inside the virus capsid obtained from the bacteriophage P22. CYP and coat protein were both heterologously expressed in E. coli. The VLPs with enzymatic activity were covered with polyethylene glycol that was functionalized in its distal end with folic acid in order to be recognized by folate receptors exhibited on tumor cells. The capacity of biocatalytic VLPs to be recognized and internalized into tumor cells is demonstrated. The VLP-treated cells showed enhanced capacity for the transformation of the pro-drug tamoxifen, which resulted in an increase of the cell sensitivity to this oncological drug. In this work, the potential use of biocatalytic VLPs vehicles as a delivery system of medical relevant enzymes is clearly demonstrated. In addition to cancer treatment, this technology also offers an interesting platform as nano-bioreactors for intracellular delivery of enzymatic activity for other diseases originated by the lack of enzymatic activity.
化疗中使用的大多数药物都需要通过细胞色素P450（CYP）酶催化的转化作用来激活。在这项研究中，设计了含有CYP活性、免疫惰性且经过功能化修饰以便被人宫颈癌细胞和人乳腺腺癌细胞识别的噬菌体P22病毒样颗粒（VLP）。CYP被包裹在从噬菌体P22获得的病毒衣壳内。CYP和衣壳蛋白均在大肠杆菌中进行异源表达。具有酶活性的VLP表面覆盖有聚乙二醇，其远端用叶酸进行了功能化修饰，以便被肿瘤细胞上表达的叶酸受体识别。证明了生物催化VLP被肿瘤细胞识别并内化的能力。经VLP处理的细胞对前药他莫昔芬的转化能力增强，这导致细胞对这种肿瘤药物的敏感性增加。在这项研究中，清楚地证明了生物催化VLP载体作为医学相关酶递送系统的潜在用途。除了癌症治疗外，这项技术还提供了一个有趣的平台，可作为纳米生物反应器用于细胞内递送酶活性，以治疗因酶活性缺乏引起的其他疾病。
