Novel Tandem Biotransformation Process for the Biosynthesis of a Novel Compound, 4-(2,3,5,6-Tetramethylpyrazine-1)-4′-Demethylepipodophyllotoxin
Novel Tandem Biotransformation Process for the Biosynthesis of a Novel Compound, 4-(2,3,5,6-Tetramethylpyrazine-1)-4′-Demethylepipodophyllotoxin
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DOI：
10.1128/aem.03047-10
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发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Li, Hong-Mei
中科院分区：
文献类型：
--
作者：
Tang, Ya-Jie;Zhao, Wei;Li, Hong-Mei
根据鬼臼毒素的结构及其构效关系，开发了一种新型串联生物转化工艺，对鬼臼毒素结构进行定向修饰，定向合成新型化合物4-(2,3,5,6-四甲基吡嗪-1)-4'-去甲基表鬼臼毒素(4-TMP-DMEP)。在这个新颖的串联生物转化过程中，鬼臼毒素的起始底物通过从神农架原始森林腐殖土（中国湖北）中筛选出的赤霉赤霉SH-f13对4'位甲氧基进行脱甲基化而生物转化为4'-去甲基表鬼臼毒素（产物1）。 4'-去甲基鬼臼毒素（产物1）通过从中国科学院武汉植物园采集的八角莲属植物中筛选出的链格孢S-f6，氧化4位羟基，生物转化为4'-去甲基鬼臼毒素（产物2）。最后，将4'-去甲基鬼臼酮（产物2）与川芎嗪进行氨基转移反应，由链格孢S-f6合成目标产物4-TMP-DMEP（产物3）。与鬼臼毒素（即50%有效浓度[EC50]为529μM）相比，4-TMP-DMEP对肿瘤细胞系BGC-823（即0.11μM）的EC50显着降低了5,199倍。同时，4-TMP-DMEP对正常人近端肾小管上皮细胞系HK-2的EC50（即0.40μM）比鬼臼毒素（即0.006μM）高66倍。此外，与鬼臼毒素（即log P = 0.34）相比，4-TMP-DMEP（即log P = 0.66）的水溶性显着增强了94%。本工作首次采用鬼臼毒素通过串联生物转化工艺定向合成了具有优异抗肿瘤活性的新型化合物4-TMP-DMEP。
According to the structure of podophyllotoxin and its structure-function relationship, a novel tandem biotransformation process was developed for the directional modification of the podophyllotoxin structure to directionally synthesize a novel compound, 4-(2,3,5,6-tetramethylpyrazine-1)-4'-demethylepipodophyllotoxin (4-TMP-DMEP). In this novel tandem biotransformation process, the starting substrate of podophyllotoxin was biotransformed into 4'-demethylepipodophyllotoxin (product 1) with the demethylation of the methoxyl group at the 4' position by Gibberella fujikuroi SH-f13, which was screened out from Shennongjia prime forest humus soil (Hubei, China). 4'-Demethylepipodophyllotoxin (product 1) was then biotransformed into 4'-demethylpodophyllotoxone (product 2) with the oxidation of the hydroxyl group at the 4 position by Alternaria alternata S-f6, which was screened out from the gathered Dysosma versipellis plants in the Wuhan Botanical Garden, Chinese Academy of Sciences. Finally, 4'-demethylpodophyllotoxone (product 2) and ligustrazine were linked with a transamination reaction to synthesize the target product 4-TMP-DMEP (product 3) by Alternaria alternata S-f6. Compared with podophyllotoxin (i.e., a 50% effective concentration [EC50] of 529 mu M), the EC50 of 4-TMP-DMEP against the tumor cell line BGC-823 (i.e., 0.11 mu M) was significantly reduced by 5,199 times. Simultaneously, the EC50 of 4-TMP-DMEP against the normal human proximal tubular epithelial cell line HK-2 (i. e., 0.40 mu M) was 66 times higher than that of podophyllotoxin (i. e., 0.006 mu M). Furthermore, compared with podophyllotoxin (i. e., log P = 0.34), the water solubility of 4-TMP-DMEP (i. e., log P = 0.66) was significantly enhanced by 94%. For the first time, the novel compound 4-TMP-DMEP with superior antitumor activity was directionally synthesized from podophyllotoxin by the novel tandem biotransformation process developed in this work.