TAXOL AND TAXANES FROM PLANT TISSUE CULTURE

该项目旨在发展植物组织生物技术，为 生产紫杉醇和紫杉醇外排阻滞剂以提高利用率 紫杉醇用于进一步的临床评估和克服紫杉醇耐药性 有问题。这一目标将由一个财团实现，该财团是 多学科(分析化学、生物化学、天然产物 化学、分子生物学和药理学)和多机构 (普渡大学和厄普约翰公司)。我们建议：(1)发展 红豆杉属植物的悬浮组织培养和毛状根培养 微管蛋白稳定化筛选高产菌落的研究 串联质谱仪/高效液相色谱；(3) 发展高灵敏、高选择性的串联质谱学 紫杉醇和其他紫杉烷的生产方法；(4)促进紫杉醇和 诱导子诱导紫杉烷和转化紫杉烷 各种农杆菌菌株；(5)筛选前潜在紫杉醇外流 来自组织培养的阻滞剂；(6)进行生物和物理分析- 定向分离，并阐明所需成分的结构； (7)以巴卡菌素III或其他相关紫杉烷为原料合成紫杉醇； 评价新型紫杉烷的体内抗肿瘤活性。

This project is aimed at developing plant tissue biotechnology for the production of taxol and taxol efflux blockers to improve the availability of taxol for further clinical evaluations and overcome taxol-resistance problems. This objective will be accomplished by a consortium which is multidisciplinary (analytical chemistry, biochemistry, natural product chemistry, molecular biology, and pharmacology) and multi-institutional (Purdue University and Upjohn Company). We propose to: (1) develop suspension tissue cultures and hairy root cultures of Taxus species; (2) analyze and select high yielding colonies by tubulin stabilization and tandem mass spectrometry/high performance liquid chromatography; (3) develop highly sensitive and compound-selective tandem mass spectrometric methods for taxol and other taxanes; (4) enhance production of taxol and desirable taxanes by induction with elicitors and transformation with various Agrobacterium strains; (5) prescreen potential taxol efflux blockers from tissue cultures; (6) perform biological and physical assay- directed isolation, and elucidate the structures of desirable constituents; (7) synthesize taxol from baccatin III or other related taxanes; and (8) evaluate antitumor activity of novel taxanes in vivo.