New enzyme for reductive cancer chemotherapy, YieF, and its improvement by directed evolution
New enzyme for reductive cancer chemotherapy, YieF, and its improvement by directed evolution
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DOI：
10.1158/1535-7163.mct-05-0365
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发表时间：
2006-01-01
影响因子：
5.7
通讯作者：
Matin, A
中科院分区：
文献类型：
--
作者：
Barak, Y;Thorne, SH;Matin, A
还原性前药丝裂霉素C和5-氮丙啶基-2，4-二硝基苯甲酰胺（CB 1954）在其天然形式下是无毒的，但在还原后变得具有高度毒性。它们在癌症化疗中的有效性可以通过向肿瘤递送具有改善的前药还原动力学的酶来增强。我们报告发现了一种新的前体药物还原酶，YieF，从大肠杆菌，并改善其动力学减少丝裂霉素C和CB 1954。在各种药物和酶浓度和孵育时间下，YieF衍生的酶Y 6以比野生型酶YieF和NfsA>= 5倍的效率杀死HeLa旋转细胞。用粘附的HeLa细胞和鼠伤寒沙门氏菌SL 7838细菌作为酶递送载体，与表达野生型酶的细菌相比，需要至少一个数量级更少的产生Y 6的细菌来杀死>90%的肿瘤细胞，所述野生型酶以相当的水平杀死肿瘤细胞。
Reductive prodrugs, mitomycin C and 5-aziridinyl-2,4-dinitrobenzamide (CB 1954), are nontoxic in their native form but become highly toxic upon reduction. Their effectiveness in cancer chemotherapy can be enhanced by delivering to tumors enzymes with improved prodrug reduction kinetics. We report the discovery of a new prodrug-reducing enzyme, YieF, from Escherichia coli, and the improvement of its kinetics for reducing mitomycin C and CB 1954. A YieF-derived enzyme, Y6, killed HeLa spinner cells with >= 5-fold efficiency than the wildtype enzymes, YieF and NfsA, at a variety of drug and enzyme concentrations and incubation times. With adhered HeLa cells and Salmonella typhimurium SL 7838 bacteria as enzyme delivery vehicle, at least an order of magnitude less of Y6-producing bacteria were required to kill >90% of tumor cells compared with bacteria expressing the wild-type enzymes, which at a comparable level killed