酶催化制备(R)-2-氯-1-[(R)-6-氟苯并二氢吡喃-2-基]-1-乙醇  

Synthesis of(R)-2-Chloro-1-[(R)-6-fluorochroman-2-yl]ethan-1-ol Catalized by Enzyme

作　　者：谷绪顶 王福军 蒲通 宋庆宝[3] 刘玉坤 GU Xuding;WANG Fujun;PU Tong;SONG Qingbao;LIU Yukun(Jiangsu BaJu Pharmaceutical Co.,Ltd.,Yancheng 224555;Zhejiang Chrioteer Pharmaceutical Stock Co.,Ltd.,Taizhou 317321;Zhejiang University of Technology School of Chemical Engineering,Hangzhou 310014)

机构地区：[1]江苏八巨药业有限公司,江苏盐城224555 [2]浙江车头制药股份有限公司,浙江台州317321 [3]浙江工业大学化工学院,浙江杭州310014

出　　处：《中国医药工业杂志》2018年第12期1677-1683,共7页Chinese Journal of Pharmaceuticals

基　　金：国家国际科技合作项目(2015DFR40360)

摘　　要：采用基因重组大肠杆菌表达(R)-羰基还原酶,通过优化诱导温度、碳源类型和补料速度,初步确定最佳产酶条件:诱导温度为28℃,碳源类型为葡萄糖,最佳补料速度为20 ml/h,产酶活力提高至63.1 U/g。利用葡萄糖脱氢酶耦合(R)-羰基还原酶催化制备奈必洛尔关键中间体(R)-2-氯-1-[(R)-6-氟苯并二氢吡喃-2-基]-1-乙醇,确定葡萄糖脱氢酶和羰基还原酶的最佳投酶量。当底物2-氯-1-[(R)-6-氟苯并二氢吡喃-2-基]-1-乙酮投料量为5 g时,葡萄糖脱氢酶和(R)-羰基还原酶的最佳投酶量为1 000 U和120 U,所制备目标化合物纯度98.3%,收率91%,手性HPLC纯度为99.6%,手性GC纯度为98.5%。Recombinant Escherichia coli was adopted to express (R)-carbonyl reductase,and the induction temperature,carbon source type and feeding speed were optimized for the best producation of (R)-carbonyl reductase. The optimum conditions for enzyme production were as follows:temperature:28°C,glucose as the carbon source and feeding rate:20ml/h.Under these conditions,the activity of (R)-carbonyl reductase was increased to 63.1U/g.The key intermediate of nepirolol,(R)-2-chloro-1-[(R)-6-fluorochroman-2-yl]ethan-l-ol was synthesized in the presence of glucose dehydrogenase coupled with (R)-carbonyl reductase.When the dosage of 2-chloro-l-[(R)-6-fluorochroman- 2-yl]ethan-l-one was 5g,the optimal dosage of glucose dehydrogenase and (R)-carbonyl reductase were 1000U and 120U respectively,the target compound was prepared with a yield of 91%,a purity of 98.3%,chiral HPLC purity of 99.6%,and chiral GC purity of 98.5%.

关 键 词：奈必洛尔 关键中间体 葡萄糖脱氢酶 羰基还原酶 酶催化合成 

分 类 号：P014.5[天文地球] R972[医药卫生—药品]