新型抗抑郁药的体内系统筛查方法及药代动力学研究

发布时间：2018-05-27 20:10

本文选题：新型抗抑郁药 + 超声辅助低密度溶剂分散液液微萃取　；参考：《福建中医药大学》2017年硕士论文

【摘要】：抑郁症是一种常见的精神疾病,主要特征为心境低落、思维迟缓等,严重者会出现自杀行为。新型抗抑郁药是目前临床上用于治疗抑郁症的主导药物。但随着新型抗抑郁药的使用不断增加,出现了药物滥用及被应用于暴力犯罪中等情况,此外,临床上存在药物联用,但对于药物的相互作用研究尚少。针对这些存在的问题,本课题研究了该类药物的快速定量方法、在生物体内的药物代谢情况及与其他抗精神病药物联用的体内相互影响,为新型抗抑郁药的用药安全、法医鉴定、基础研究及临床研究提供技术手段和参考依据。具体的研究内容如下:1超声辅助-低密度溶剂-分散液液微萃取GC/MS法同时测定人全血中12种新型抗抑郁药和2种常用抗精神病药以超声辅助-低密度溶剂-分散液液微萃取(UA-LDS-DLLME)技术进行人全血的样品前处理,建立了 12种新型抗抑郁药与2种常用的抗精神病药物同时定量的方法。12种新型抗抑郁药分别为诺氟西汀、氟西汀、氟伏沙明、阿戈美拉汀、米氮平、吗氯贝胺、美利曲辛、去甲米氮平、马普替林、舍曲林、西酞普兰和帕罗西汀,2种抗精神病药物为氯氮平和氟哌啶醇。对人全血的UA-LDS-DLLME前处理方法进行优化,确立了最优条件:以100μL甲苯为萃取剂,加入100 μLpH12的氨水,超声3min。GC/MS的测定条件为:色谱柱:DB-5ms(0.25mm×3.0m×0.25 μm),不分流进样,进样时间为1min,进样口温度280℃;柱箱升温程序:100℃(1 min)_20℃/min_20℃(5min)_8℃/min_300℃(5min)。EI电子轰击源,70eV;离子源温度23℃;接口温度300℃,.溶剂延迟4min;选择离子扫描模式。选用诺氟西汀-D6和帕罗西汀-D6为内标。方法学验证结果表明该方法具有专属性,14个待测化合物分别在15-1500 ng/mL或5-500 ng/mL范围内线性良好(r20.99),日内(n=5)及日间(n=15)的准确度和精密度均符合规定要求(准确度在±15%以内,精密度小于15%),大部分化合物的提取回收率在50%以上,稳定性良好。该方法已成功应用于实际案例中。2基于液相色谱-高分辨质谱(LC-HRMS)的新型抗抑郁药美利曲辛在大鼠体内的代谢研究建立了基于UHPLC/LTQ-Orbitrap的LC-HRMS联用方法,通过结合精确质量数和多级碎片离子信息,研究抗抑郁药美利曲辛在大鼠体内的代谢产物与代谢途径。以美利曲辛灌胃给药大鼠,收集大鼠给药前及给药后的3、6、9、12、24h各时间段的尿液用于分析,分别采用固相萃取和液液萃取进行样品前处理,最终在大鼠尿液中找到美利曲辛及其Ⅰ相和Ⅱ相代谢物共30种,并初步推断了美利曲辛在大鼠体内的代谢途径主要为去甲基和羟基化。3大鼠血浆中度洛西汀和喹硫平的LC-MS/MS法测定及药代动力学研究建立了同时测定大鼠血浆中度洛西汀和喹硫平的LC-MS/MS定量方法,采用蛋白沉淀法处理血浆。采用EclipseXDB-C18柱,流动相A为含2 mmol/L甲酸铵和0.1%甲酸的水相,流动相B为含2mmol/L甲酸铵和0.1%甲酸的乙腈有机相,采用梯度洗脱,分析时间为6.0 min。质谱采用MRM模式,化合物与内标的离子对分别为:度洛西汀m/z 298.1→154.1,喹硫平 m/z 384.4→253.2,内标(氟哌啶醇)m/z 376.2→165.2。经方法学验证,该法具有专属性,度洛西汀和喹硫平的定量下限分别为0.500 ng/mL和1.00 ng/mL,在0.500～100 ng/mL和1.00～200 ng/mL范围内线性关系良好,准确度和精密度均在允许的范围内,提取回收率均会87.7%,总体内标归一化的基质效应为1.03和0.94,能满足生物样品的分析要求。将此方法成功应用于度洛西汀与喹硫平联合用药的药代动力学研究,将SD大鼠随机分为3组,每组6只,分别进行单一给药度洛西汀、单一给药喹硫平及联合给药度洛西汀和喹硫平,通过药代动力学参数来评价合并用药后的相互影响。度洛西汀的单一给药组和联合用药组的Cmax和AUC0-∞分别为34.4 ± 5.6 ng/mL、372.7 ± 105.9 h-ng/mL和55.9 ± 13.0 ng/mL、544.1 ± 56.0 h·ng/mL。喹硫平的单一给药组和联合用药组的Cmax和AUC0-∞分别为 18.1 ±14.6 ng/mL、35.2±35.0h·ng/mL 和 22.8± 15.5 ng/mL、41.2±21.1 h·ng/mL;经过配对t检验,喹硫平的单一给药组和联合给药组的Cmax和AUC0-∞的,P值均大于0.05,说明联合用药对喹硫平的药代动力学无明显影响;而度洛西汀的两组间的Cmax和AUC0-∞的P值分别为0.016和0.039,AUC0-t的P值为0.001,说明联合用药对度洛西汀的药代动力学有明显影响,喹硫平能增加度洛西汀在大鼠体内的血药浓度,其药代动力学参数Cmax和AUC明显增大。
[Abstract]:Depression is a common mental illness, characterized by low mood, slow thinking, and serious mental retardation. The new antidepressant is the dominant drug used in the treatment of depression, but with the increasing use of new antidepressants, drug abuse and the medium of use of violent crimes have emerged. In addition, there are clinical drugs in combination, but there are few studies on the interaction of drugs. In view of these problems, we have studied the rapid quantitative methods of the drugs, the drug metabolism in the organism and the interaction with other antipsychotic drugs in vivo, and the safety of the new antidepressant drugs and the forensic evaluation. The basic research and clinical research provide technical means and reference basis. The specific research contents are as follows: 1 ultrasonic assisted low density solvent dispersible liquid liquid microextraction GC/MS method for simultaneous determination of 12 new antidepressants and 2 common antipsychotics in human whole blood with ultrasonic assisted low density solvent dispersible liquid microextraction (UA-LDS-DLLME) technique 12 new antidepressants, 12 new antidepressants and 2 commonly used antipsychotics, were established for the simultaneous determination of the whole blood samples. The new antidepressants were norfluoxetine, fluoxetine, Hide Saki, ametaline, Mi Danping, mopectrine, and meritactyine, to Krabi, mopecin, sertraline, and citalopram. And Pa Rossi Dean, 2 kinds of antipsychotic drugs were clozapine and haloperidol. The optimal conditions for UA-LDS-DLLME pretreatment of human whole blood were optimized. The optimum conditions were established: 100 u L toluene as extractant and 100 mu LpH12 of ammonia water. The ultrasonic 3min.GC/MS was determined by the chromatographic column: DB-5ms (0.25mm x 3.0m x 0.25 mu), without diversion and injection. The room temperature was 1min, the inlet temperature was 280 C; the column box heating program: 100 C (1 min) _20 C /min_20 C (5min) _8 C /min_300 (5min).EI electron bombardment source, 70eV, the temperature of the ion source 23, the interface temperature 300, the solvent delay 4min; the selective ion scanning mode. The method has special properties. The 14 compounds to be tested are linear in the range of 15-1500 ng/mL or 5-500 ng/mL respectively (r20.99). The accuracy and precision of intraday (n=5) and day (n=15) are all in accordance with the requirements (accuracy is within + 15%, precision is less than 15%). The extraction recovery of most compounds is above 50%, and the stability is good. The method has been proved to be good. Successfully applied to practical cases,.2 based on the metabolism of a new antidepressant,.2, based on liquid chromatography high resolution mass spectrometry (LC-HRMS), established a LC-HRMS combined method based on UHPLC/LTQ-Orbitrap. By combining accurate mass number and multilevel fragment ion information, the antidepressant, Meili, was studied in rats. The metabolites and metabolic pathways were administered to the rats by gastric perfusion, and the urine of the rats was collected before and after the administration of the 3,6,9,12,24h. Solid phase extraction and liquid extraction were used for sample pretreatment respectively. In the end, 30 kinds of metabolites were found in the urine of rats and their phase I and II phase. The metabolic pathways of milleine in rats were determined mainly by the LC-MS/MS determination and pharmacokinetics of moderate levioxetine and quetiapine in the plasma of.3 rats. A quantitative method for the simultaneous determination of the moderate levels of luoxetine and quetiapine in rat plasma was established, and plasma was treated by protein precipitation method, and EclipseXDB-C was used for the treatment of plasma. The 18 column, the mobile phase A is the aqueous phase of 2 mmol/L ammonium formate and 0.1% formic acid, and the mobile phase B is the organic phase of acetonitrile containing 2mmol/L formate and 0.1% formic acid. The gradient elution is used, the analysis time is 6 min. mass spectrometry using MRM mode, the compound and the internal standard ion pair are respectively dloxetine m /z 298.1 - 154.1, quetiapine m/z 384.4 to 253.2, internal standard (fluorine) Piperidine) m/z 376.2 to 165.2. is verified by square jurisprudence. This method has special properties. The quantitative lower limit of duloxetine and quetiapine is 0.500 ng/mL and 1 ng/mL respectively. The linear relationship is good in the range of 0.500 ~ 100 ng/mL and 1 to 200 ng/mL. The accuracy and precision are within the allowable range, and the recovery rate will be 87.7% and the overall standard is classified. The matrix effect of one chemical was 1.03 and 0.94, which could satisfy the analysis requirements of biological samples. The method was successfully applied to the pharmacokinetics study of duloxetine and quetiapine. The SD rats were randomly divided into 3 groups, 6 rats in each group, single administration of quinoline, single dose quetiapine and combined duloxetine and quetiapine, The Cmax and AUC0- infinity of the single drug group and the combination group of duloxetine were 34.4 + 5.6 ng/mL, 372.7 + 105.9 h-ng/mL and 55.9 + 13 ng/mL, 544.1 + 56 h ng/mL. quetiapine, and Cmax and AUC0- infinity of the combined drug group respectively. 18.1 + 14.6 ng/mL, 35.2 + 35.0h / ng/mL and 22.8 + 15.5 ng/mL, 41.2 + 21.1 H. Ng/mL. After paired t test, the Cmax and AUC0- infinity of the quetiapine group and the combined administration group were all greater than 0.05, indicating that the combined medication had no obvious effect on the pharmacokinetics of quetiapine, while the Cmax and AUC0- infinity between the two groups of duloxetine were found. The values were 0.016 and 0.039 respectively, and the P value of AUC0-t was 0.001, which indicated that the combined use of drugs had a significant effect on the pharmacokinetics of duloxetine. Quetiapine could increase the concentration of blood drug in rats, and the pharmacokinetic parameters of Cmax and AUC were significantly increased.
【学位授予单位】：福建中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96

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