Vasilev Nikolay P, Julsing Mattijs K, Koulman Albert, Clarkson Cailean, Woerdenbag Herman J, Ionkova Iliana, Bos Rein, Jaroszewski Jerzy W, Kayser Oliver, Quax Wim J
Department of Pharmaceutical Biology, University of Groningen, Groningen Research Institute of Pharmacy (GRIP), Antonius Deusinglaan 1, NL-9713 AV Groningen, The Netherlands.
J Biotechnol. 2006 Nov 10;126(3):383-93. doi: 10.1016/j.jbiotec.2006.04.025. Epub 2006 Jun 5.
Biotransformation of deoxypodophyllotoxin to epipodophyllotoxin by three major human hepatic enzymes, CYP1A2, CYP2C9 and CYP3A4, heterologously expressed in E. coli DH5alpha, was investigated. It was shown that CYP3A4 catalysed the hydroxylation of deoxypodophyllotoxin into epipodophyllotoxin in yields up to 90%. The structure of the metabolite was determined using HPLC-MS and HPLC-SPE-NMR techniques. There was no detectable production of epipodophyllotoxin or podophyllotoxin by CYP1A2 and CYP2C9 enzymes. The CYP3A4 enzyme shows a distinctly different reactivity to deoxypodophyllotoxin compared to the semi-synthetic derivatives etoposide and teniposide, which are degraded by 3-O-demethylation. These findings demonstrate a novel system for the production of 2,7'-cyclolignans, starting from the easily accessible deoxypodophyllotoxin.
研究了在大肠杆菌DH5α中异源表达的三种主要人肝酶CYP1A2、CYP2C9和CYP3A4将脱氧鬼臼毒素生物转化为表鬼臼毒素的过程。结果表明，CYP3A4催化脱氧鬼臼毒素羟基化生成表鬼臼毒素，产率高达90%。使用HPLC-MS和HPLC-SPE-NMR技术确定了代谢物的结构。CYP1A2和CYP2C9酶未检测到表鬼臼毒素或鬼臼毒素的产生。与通过3-O-去甲基化降解的半合成衍生物依托泊苷和替尼泊苷相比，CYP3A4酶对脱氧鬼臼毒素表现出明显不同的反应性。这些发现证明了一种从易于获得的脱氧鬼臼毒素开始生产2,7'-环木脂素的新系统。
