去甲斑蝥新型衍生物N-14NCTDA大鼠肝微粒体酶代谢  

Metabolic studies of N-14NCTDA,a new norcantharidin derivative,in rat liver microsomes

作　　者：潘振东[1] 杨柳[1] 唐星[1] 蔡翠芳[1] 

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2015年第12期956-961,966,共7页Journal of Shenyang Pharmaceutical University

摘　　要：目的初步阐明十四烷基去甲斑蝥素酰亚胺(tetradecyl derivative of norcantharidin,简称N-14NCTDA)肝微粒体酶代谢情况,鉴定该药物体外Ⅰ相代谢产物,并推测其主要代谢途径。方法以超速离心法制备大鼠肝微粒酶并以二辛可酸(butyleyanoacrylate,BCA)法测定微粒体蛋白浓度。药物与肝微粒体在一定条件下孵育,使用飞行时间质谱(UPLC/Q-TOF MS)进行分析。结果制备的大鼠肝微粒体酶蛋白质量浓度约为6.4 g·L^(-1)。代谢研究结果显示,孵育后反应液中主要检测到3个代谢产物,其分子离子峰相对分子质量分别为380.280 6、378.264 8和394.259 7,分别对应原型药羟基化、醛基化及羧化产物。结论 UPLC/Q-TOF MS方法适用于N-14NCTDA和其代谢产物的定性鉴别。体外初步代谢结果表明,N-14NCTDA的酰亚胺键难以在肝脏水解释放出NCTD,其主要以末端烷基氧化的方式进行Ⅰ相代谢。Objective To study the in vitro metabolic characteristics of N-14 NCTDA with rat liver microsomes,and identify its phaseⅠmetabolits and infer its major metabolic pathways. Methods The rat liver microsomes were prepared by differential ultracentrifugation and then the enzyme activity was determined by BCA method. In this study,an ultra high performance liquid chromatography-time of flight mass spectrometric method（ UPLC / Q-TOF M S） was developed to characterize the metabolites of N-14 NCTDA after incubation with liver microsomes. Results The protein concentration of prepared rat liver microsomes was determined to be 6. 4 g·L^（-1）. Three major metabolites were found and identified in this incubation system by UPLC / Q-TOF M S method. The molecular weights of its molecular ion peaks were found to be 380,378 and394,respectively. The results demonstrated that the in vitro metabolic pathway was that the end alkyl was first oxidized to hydroxyl group,then further oxidized to a aldehyde group and carboxyl group. Conclusions A rapid and sensitive UPLC / Q-TOF M S method has been established for qualitative study of N-14 NCTDA and its metabolites. The preliminary in vitro metabolic study indicated that the imide bond of N-14 NCTDA was hard to be hydrolyzed in rat liver microsome incubation system,and the oxidation of end alkyl group was the major pathway of phaseⅠmetabolism of N-14 NCTDA.

关 键 词：去甲斑蝥素 衍生物 肝微粒体酶 飞行时间质谱 体外代谢 

分 类 号：R917[医药卫生—药物分析学]