【作者】 宋乃宁；

【导师】 刘昌孝；

【作者基本信息】 天津大学 ， 新药评价学， 2009， 博士

【摘要】 本文进行了创新的抗癌药物5-溴粉防己碱(溴泰君)的药代动力学、毒代动力学以及探索性地应用代谢组学方法研究其毒性标志物。本研究是国家973计划“基于功能基因组学的创新药物研究”课题(2004BC518902)和天津市科技发展计划重点项目“代谢组学用于中药安全性和作用机理研究”课题(05YFJZJC01100)的基础研究内容之一。从以下四方面获得了具有科学意义和应用价值的研究结果:1.建立了大鼠血浆中粉防己碱测定的液相色谱/质谱联用法,该方法快速、灵敏、专署性强,并应用此方法进行粉防己碱单剂量口服给药后在大鼠体内的药代动力学研究,结果显示粉防己碱口服给药的体内代谢过程符合一室模型;2.建立了大鼠血浆中溴泰君的液相色谱/质谱联用测定方法,并将该方法应用于溴泰君单剂量口服给药后在大鼠体内的药代动力学研究,结果显示溴泰君口服给药后在雌雄大鼠体内代谢存在差异,体内代谢过程均符合一室模型;3.分别进行了粉防己碱和溴泰君两种药物多剂量单次给药、单剂量多次口服给药在大鼠体内的毒代动力学研究,以及毒理学研究。单剂量毒代动力学研究结果显示粉防己碱和溴泰君在致毒剂量下,在大鼠体内的代谢动力学参数均发生变化;多剂量毒代动力学研究记录了两种药物在体内产生毒性的血药浓度经时变化规律,结果显示,高剂量连续给药后,由于毒性作用,粉防己碱和溴泰君在大鼠体内的代谢动力学行为均发生改变。毒理学研究通过血液生化指标的测定和组织病理切片的观察表明,粉防己碱在毒性剂量下,主要表现为肝脏、肺和脾的损伤,停药6天后,病变并未减轻;溴泰君在毒性剂量下,同样主要表现为肝脏、肺和脾的损伤,其中肝脏毒性作用程度轻于粉防己碱,停药后,病变有所减轻但无统计学意义;4.分别进行粉防己碱和溴泰君两种药物的初步探索性代谢组学研究,利用液相色谱/质谱联用法的测定方法,使用XCMS统计软件进行数据的处理分析,考察机体分别对粉防己碱和溴泰君的毒性作用产生应激反应所造成的血浆、尿液中内源性代谢产物的经时变化规律;寻找两种药物毒性作用的可能的生物标志物离子,分别在血浆、尿液中找到粉防己碱毒性的生物标志物离子10和18个,溴泰君的生物标志物离子16和17个。结合毒代动力学与毒理学研究结果,验证两种药物的毒性靶器官为肝脏,并为今后进一步毒性机制研究提供信息。更多还原

【Abstract】 In the present study, traditional Chinese medicine tetrandrine and its 5-bromized derivative bromotetrandrine were studied and evaluated through the combination of pharmacokinetics, toxicokinetics and metabonomincs method by which toxic markers were searched. This study is an important part of National 973 Plan Foundation of China (No. 2004BC518902) and Tianjin Development Plan of Science and Technology (05YFJZJC01100). Results with both scientific and practical significance were listed as follows:1. A rapid and sensitive liquid chromatography-tandem mass spectrometric method (LC/MS/MS) for the determination of tetrandrine in rat plasma has been developed, fully validated and successfully applied to a pharmacokinetic study in Sprague-Dawley (S.D.) rats after a single oral administration. The result showed that the metabolism of tetrandrine in both male and female rats was best fitted to one-compartment model.2. A rapid and sensitive liquid chromatography-tandem mass spectrometric method (LC/MS/MS) for the determination of bromotetrandrine in rat plasma has been developed, fully validated and successfully applied to a pharmacokinetic study in Sprague-Dawley (S.D.) rats after a single oral administration. The result exhibited sexual difference in the pharmacokinetics of bromotetrandrine between male and female rats, though its metabolism in both male and female rats was best fitted to one-compartment model.3. Single-dose and multi-dose toxicokinetic study, as well as toxicological study of both tetrandrine and bromotetrandrine were done in S.D. rats for evaluating kinetic properties and repeated dose safety for the two drugs. Results of single-dose toxicokinetic studies showed that at the dose that might induce toxicity, kinetic parameters of both tetrandrine and bromotetrandrine changed in rats. Multi-dose toxicokinetic studies recorded the changing states of tetrandrine and bromotetrandrine concentration in rat plasma, and demonstrated that after multiple administration of the two drugs at the dose that might induce toxicity, the metabolism of tetrandrine and bromotetrandrine were both changed in rats. Determination of blood biochemical parameters and tissue pathological slices results showed that both tetrandrine and bromotetrandrine exhibited liver, spleen and lung damage, the degree of all the damage was not alleviated after 6 days’recovery. 4. In the present study, we also did some primary metabonomic study for both tetrandrine and bromotetrandrine. LC-MS was developed for urine and plasma sample determination, and XCMS was exploited for data handling and analysis to research time-related changes of endogenous metabolic responses in rat urine and plasma to tetrandrine and bromotetrandrine stimuli, in order to find biomarkers in biochemical matrix. As a result 10 and 18 promising biomarkers were found in plasma and urine for tetrandrine, respectively. And 16 and 17 for bromotetrandrine in plasma and urine, respectively. Combined with the results of toxicokinetic studies, it was further confirmed that both tetrandrine and bromotetrandrine exhibited hepatotoxicity.更多还原