色谱 ›› 2012, Vol. 30 ›› Issue (02): 165-169.DOI: 10.3724/SP.J.1123.2011.10021

• 研究论文 • 上一篇    下一篇

工业色谱法分离制备7-木糖基-10-去乙酰紫杉醇酶解产物10-去乙酰紫杉醇

刘兴宝, 栾宏伟, 葛广波, 侯琨, 杜逊甫, 杨凌*   

  1. 中国科学院大连化学物理研究所, 辽宁 大连 116023
  • 收稿日期:2011-10-20 修回日期:2011-12-04 出版日期:2012-02-28 发布日期:2012-03-22
  • 通讯作者: 杨凌,博士,研究员,博士生导师,主要从事药用资源开发及早期成药性研究导向下的新药发现. Tel: (0411)84379317,
  • 基金资助:

    国家重大科学计划项目(2012CB910603和2010CB912701)和国家自然科学基金项目(20735005、20875101和20905077)

Industrial preparative chromatography purification of 10-deacetylpaclitaxel, the enzymatic product of 7-xylosyl-10-deacetylpaclitaxel

LIU Xingbao, LUAN Hongwei, GE Guangbo, HOU Kun, DU Xunfu, YANG Ling*   

  1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
  • Received:2011-10-20 Revised:2011-12-04 Online:2012-02-28 Published:2012-03-22

摘要: 基于工业色谱法分离制备抗癌药物紫杉醇的半合成前体10-去乙酰紫杉醇(10-DAP)。7-木糖基-10-去乙酰紫杉醇(10-DAXP)在我国特有红豆杉品系(中华红豆杉)枝叶中含量丰富,以其为原料可制备紫杉醇最理想的半合成前体——10-DAP。本研究以部分纯化后的7-木糖基-10-去乙酰紫杉烷为原料,通过β-木糖苷酶水解该粗提物中的主要成分10-DAXP及其两个微量类似物7-木糖基-10-去乙酰三尖杉宁碱(10-DAXC)和7-木糖基-10-去乙酰紫杉醇C(10-DAXP C),脱去其C-7位上的木糖基,水解产物采用大孔吸附树脂吸附,正相快速柱分离和反相制备色谱分离,可获得高纯度的目标物10-DAP,产物纯度为96%,整个工艺的收率大于75%。该方法适合以10-DAXP为原料大规模制备紫杉醇的半合成前体化合物10-DAP,为工业化生产紫杉醇开辟了一条新途径。

关键词: 10-去乙酰紫杉醇, 7-木糖基-10-去乙酰紫杉醇, 7-木糖基-10-去乙酰紫杉烷, 分离纯化, 制备色谱, 紫杉醇

Abstract: A scheme for industrial preparative chromatography purification of 10-deacetylpaclitaxel (10-DAP), the semi-synthesized precursor of anticancer drug paclitaxel was developed. 7-Xylosyl-10-deacetylpaclitaxel (10-DAXP) is the most abundant constitute in the needles of Taxus Chinensis, a specific yew species distributed in China. 10-DAXP has been recognized as a good material to convert into 10-DAP, the most ideal precursor of paclitaxel. The partially purified extract from yew needles which mainly contains 10-DAXP (>60%) and other two minor 7-xylosyl-10-deacetyltaxanes including 7-xylosyl-10-deacetylcephalomannine (10-DAXC) and 7-xylosyl-10-deacetylpaclitaxel C (10-DAXP C), was used as the starting material. The total scheme can be divided into four steps. Firstly, the starting material was hydrolyzed by β-xylosidase to remove the C-7 xylosyl group completely; and then the hydrolyzed products mainly containing 10-DAP were eluted on a column packed with resin to get crude 10-DAP (with the purity of 20.5%) with high yield (96.3%). The crude 10-DAP was purified by a column packed with normal phase, and then by a reversed-phase preparative chromatography with ODS as the solid phase. After these two steps, the purity of the aim product 10-DAP was 96% with the overall yield of 79.7%. This novel scheme was suitable for large-scale purification of 10-DAP from 10-DAXP.

Key words: 7-xylosyl-10-deacetylpaclitaxel, 7-xylosyl-10-deacetyltaxane, isolation and purification, paclitaxel, 10-deacetylpaclitaxel, preparative chromatography