东北红豆杉细胞培养生产紫杉醇的调控研究

研究了诱导子、前体及抑制剂的协调作用对东北红平杉生产紫杉醇的影响。结果表明,向培养基中加入80mg/L水到、80mg/L茉莉酸甲酯、0.5mmol/L乙酸钠、2mmol/L苯丙氨酸、0.5mmol/L丝氨酸、0.1mmol/L甘氨酸、10mg/L肉桂酸、0.5mmol/L苯甲酸钠、5mmol/L丙酮酸钠、10mg/L氯化氯胆碱、和1mg/L赤霉酸可使紫杉醇含量提高368.65%。并且证明交互作用对紫杉醇合成有显著作用。     

代谢调节剂对紫杉醇和Taxuyunnanine C生物合成的调控作用

研究了诱导子、前体和抑制剂对东北红豆杉生产紫杉醇和taxuyunnanine C的影响。结果表明,诱导子在第12d添加,前体和抑制剂在第15d添加能有效地提高紫杉醇和taxuyunnanine C的含量。水杨酸与氯化氯胆碱的交互作用对紫杉醇的合成有很大影响,水杨酸与赤霉酸的交互作用对taxuyunnanine C的合成有很大影响。     

真菌诱导子对中国红豆杉生产紫杉醇优化模型研究

采用均匀设计,研究了真菌诱导子F5质量浓度,添加阶段与处理时间等对中国红豆杉细胞生产紫杉醇（Taxol）的综合效应,建立了以紫杉醇产量为目标函数的数学模型,借助模型,研究了各因子及其交互作用对紫杉醇含量的影响,获得了第15d加入质量浓度为0.64mg/L的真菌诱导子F5,处理29d为最佳的工艺组合。     

烟曲霉TMS-26产紫杉醇补料分批发酵条件初步探索

内生真菌发酵法是解决紫杉醇药源短缺问题的有效途径之一。本研究以摇瓶分批发酵为基础,进行摇瓶补料分批发酵研究,探究了苯丙氨酸、甘氨酸、苯甲酸钠乙酸钠混合液、3,5-二硝基水杨酸、H₂O₂、CuSO₄在发酵周期（13d）中,不同添加时间点对TMS-26菌体量及紫杉醇产量的影响,发现在第8天添加苯丙氨酸、甘氨酸、3,5-二硝基水杨酸时,其产量分别达到了(664.80±40.34)µg/L、(628.72±30.44)µg/L、(641.36±19.62)µg/L;在第9天添加CuSO₄时,其产量达到了(697.46±15.76)µg/L;在第10天添加H₂O₂、苯甲酸钠乙酸钠混合液,其产量分别达到了(615.78±36.28)µg/L、(792.54±10.04)µg/L。在摇瓶补料分批发酵研究结果的基础上,进行了5L罐发酵工艺放大研究,探究了前体和诱导子通过进行一次补加和恒速补加的方式对Aspergillus fumigatus TMS-26菌体量及紫杉醇产量的影响,结果表明恒速补加苯丙氨酸乙酸钠混合液,紫杉醇产量达到了746.17µg/L。通过本次研究,优化了TMS-26产紫杉醇摇瓶补料分批发酵和5L罐发酵工艺,为后续实现紫杉醇工业化生产奠定基础。     

红豆杉紫杉醇提取方法的建立

目的：研究粉碎粒度、提取溶剂、提取时间、料液比等因素对红豆杉中紫杉醇得率的影响,建立一种高效、快速的提取和检测方法。方法：采用有机溶剂-超声波提取法从红豆杉叶片中提取紫杉醇,用HPLC法对其进行鉴定,用标准曲线法进行定量分析。结果：将原料于95％烘箱内干燥至恒重,粉碎至100目,以95％甲醇为提取剂,于60℃提取20h,料液比为1：10,紫杉醇得率为0．086mg／g。结论：该法简单、快速、准确、适应性强,为紫杉醇的提取与检测提供了有效工具。     

紫杉醇的结构修饰及构效关系

本文总结了近年来对紫杉醇进行结构修饰所得到的近80种紫杉醇类似物的合成方法及其构效关系.     

紫杉醇产生菌分离的研究

报道了1993年从东北红豆杉树(Taxuscuspidata Sieb.et     

Intratesticular injection as a method to assess the potential toxicity of various agents and to study mechanisms of normal spermatogenesis

To better understand, to optimize, and to validate the technique of intratesticular (i.t.) injection, several parameters related to i.t. injection were examined. Volumes exceeding 50 μl could be injected i.t.; however, testes frequently became excessively turgid and backflow of injected fluids occurred. Thus, a volume of 50 μl or less was deemed optimal for injection. To determine the rate of distribution of substances throughout the testis, trypan blue was injected i.t. near the caudal pole of the testis, and the movement of dye was monitored. Within 2 min, the dye had spread approximately 1 cm from the site of injection, and in 5 min it had spread twice that distance. In 2 h, the dye had become distributed throughout the testis except at its extreme cranial pole. Seminiferous tubules did not take up dye, indicating that the spread of dye was via peritubular lymphatics. Seminiferous tubule histology appeared virtually unaffected by i.t. injection, even at regions adjacent to the site of injection, when a sterile 26-gauge or smaller bore needle was utilized. To determine disappearance from the testis, radiolabeled inulin was injected i.t. Half time for absorption was achieved at 1.75 h. Potential vehicles were expolored in which compounds with a variety of physical properties could be injected. Gum tragacanth, normal saline, ethylene glycol, dimethyl sulfoxide (DMSO) mixed 1:1 with normal saline, sesame oil, and propylene glycol were found to be suitable injection vehicles, whereas ethanol, dissolved in normal saline in concentrations as low as 0.5% was found unsuitable. To assess vehicle efficiency, various vehicles were utilized with a known testicular toxin (taxol) and injected into one testis, and the histology was compared with the contralateral testis injected with vehicle alone. All vehicles, found suitable above, allowed dispersion of taxol to influence areas distant from the site of injection. Intratesticular injection assesses the potential of agents to directly affect the testis, and systemic metabolism is avoided. Their rapid spread throughout the lymphatics of the testes allows seminiferous tubules to be exposed to agents in innocuous vehicles more rapidly and in higher concentration than is often possible when using systemic injections.