【作者】 王晓琳；

【导师】 王唯红； 刘会臣；

【作者基本信息】 山东大学 ， 药物分析， 2011， 硕士

【摘要】 硫辛酰维格列汀为首次合成的二肽基肽酶Ⅳ抑制剂类化合物,初步药理实验证明其具有降低大鼠血糖的作用,本文旨在研究其临床前药代动力学性质及代谢情况,以揭示药物体内处置情况,为药物的进一步研究开发奠定基础。1.本文建立了测定大鼠血浆中硫辛酰维格列汀的液相色谱-串联质谱方法,该方法选择性好,灵敏度高。采用液液萃取法由血浆中提取待测物与内标苯海拉明,色谱分析采用Shim-pack VP-ODS柱(5μm粒径,150 mmx2.0mm I.D.),流动相为甲醇-5 mM乙酸铵(80：20,V/V),流速为0.2 mL/min。通过电喷雾离子化质谱的正离子多反应监测模式进行检测,待测物与内标的定量离子对分别为492.3→189.0和m/z 256.2→167.3。在2.00～500 ng/mL浓度范围内线性良好,定量下限为2.00 ng/mL,批内、批间精密度(以相对标准偏差表示)均小于15%,准确度(以相对误差表示)在±15%以内。2.该方法成功应用于大鼠血浆中硫辛酰维格列汀的测定及药代动力学研究。大鼠禁食过夜后,分三剂量(25,50和100 mg/kg)灌胃给予硫辛酰维格列汀,峰浓度为63.9～296μg/L,药时曲线下面积为260～1214μg/L,两者均与剂量成比例。达峰时为1.25～1.84 h,表观清除率为100 L/h/kg左右。在25～100 mg/kg的剂量范围内,大鼠单次口服硫辛酰维格列汀后呈现线性药代动力学特征,硫辛酰维格列汀呈现很高的表观清除率(86～105L/h/kg)和表观分布容积(364～711L/kg),说明硫辛酰维格列汀的生物利用度可能较低,或者其在大鼠体内分布广泛或在组织中聚积,导致血药浓度较低。3.本文利用液相色谱与三重四级杆质谱研究了硫辛酰维格列汀在大鼠体内的代谢情况况,在尿液与胆汁中发现五种代谢物。在单次给予硫辛酰维格列汀之后,收集大鼠尿液与胆汁样本,用固相萃取法提取,经色谱分离后,收集相应时间段的流出物,浓缩之后进行分析定性。所发现的五种代谢物情况如下：M2为金刚烷环羟基化的代谢物,M5为1,2-二硫戊环开环后双甲基化的代谢物,M7为硫辛酸侧链羟基化的代谢物,M8为硫辛酸侧链双羟基化的代谢物,M9为硫辛酸侧链与金刚烷环同时羟基化的代谢物。更多还原

【Abstract】 Lipoyl vildagliptin is a novel dipeptidyl peptidase IV (DPP IV) inhibitor synthesized firstly, and it had been found that lipoyl vildagliptin could significantly lower the plasma glucose levels in the diabetic rats after oral administration. The pharmacokinetics and metabolism of lipoyl vildagliptin were studied in this paper, predicting the overall pharmacokinetic and metabolism profile of this novel compound, and laying foundations for the further investigation.1. A selective and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated for the determination of lipoyl vildagliptin in rat plasma. The analyte and internal standard (I.S.), diphenhydramine, were extracted from plasma using liquid-liquid extraction and separated on a Shim-pack VP-ODS column (5μm,150 mmx2.0 mm I.D.) using methanol-5 mM ammonium acetate (80:20, v/v) as mobile phase at a flow rate of 0.2 mL/min. Detection was achieved with positive electrospray ionization mass spectrometry using multiple reaction monitoring (MRM) of the transitions at m/z 492.3→189.0 and m/z 256.2→167.3 for analyte and I.S., respectively. The assay was linear over the concentration range of 2.00-500 ng/mL for analyte. The lower limit of quantification (LLOQ) was 2.00 ng/mL in rat plasma with intra-and inter-day precision as relative standard deviation (R.S.D.)<15% and accuracy as relative error (R.E.) within±15%.2. The method was successfully applied to determine lipoyl vildagliptin in rat plasma, and the pharmacokinetics of lipoyl vildagliptin was studied in rats after oral administration for developing it as an antidiabetic agent. After an overnight fasting, rats were orally given lipoyl vildagliptin. Following a single oral dose of 25,50, and 100 mg/kg, the maximum plasma concentration (Cmax) values (63.9-296μg/L) and the area under the plasma concentration-time curve (AUCo-∞) values (260～1214μg/h/L) were proportional to the doses, the time to reach Cmax (Tmax) values were from 1.25 to 1.84 h, and the oral apparent clearance (CL/F) values were around 100 L/h/kg.In rats, lipoyl vildagliptin displayed linear pharmacokinetics after a single oral dose in the range of 25～100 mg/kg. Lipoyl vildagliptin might have very high CL/F values (86～105 L/h/kg) and oral apparent volume of distribution (Vd/F) values (364-711 L/kg), which indicated that the bioavailability of this drug might be low or lipoyl vildagliptin might distribute extensively or accumulate in tissues in view of its high liposolubility.3. The metabolism of lipoyl vildagliptin in rats has been studied using a triple quadrupole mass spectrometer coupled with LC, and five phase I metabolites in urine or bile were identified. After a single oral dose of lipoyl vildagliptin, the rat urine and bile sample were collected, processed using solid-phase extraction, separated by LC, and then the corresponding effluent was collected, concentrated, and analyzed. The following metabolites were found in rats, including M2 (monohydroxylation at the adamantyl ring), M5 (S-methylation resulting from ring opening of the 1,2-dithiolane ring), M7 (monohydroxylation at the lipoic acid moiety), M8 (dihydroxylation at the lipoic acid moiety), and M9 (dihydroxylation at the lipoic acid moiety and the adamantyl ring).更多还原