伊潘立酮及其两种主要代谢产物人体内定量分析方法及药代动力学研究

发布时间：2018-05-08 12:49

本文选题：伊潘立酮 + 代谢产物P88　；参考：《吉林大学》2014年硕士论文

【摘要】：研究目的建立满足伊潘立酮（ILP）临床药代动力学研究要求的同时定量分析人的血浆样品中ILP及其代谢产物P88、P95的液相色谱-串联质谱（LC-MS/MS）方法；测定中国健康志愿者血浆样品中ILP及其代谢产物P88、P95药物浓度，绘制三种待测物的药物浓度-时间曲线，分别计算其药代动力学参数，考察其在体内的药代动力学特征，为伊潘立酮片上市提供数据支持。研究方法使用以乙腈作为洗脱剂的固相萃取方法，对人血浆中的伊潘立酮及其代谢产物P88、P95进行LC-MS/MS分析。以甲醇-10mM醋酸铵-0.1%甲酸溶液为流动相进行梯度洗脱，三种待测物及内标苯海拉明经SUPELCOAscentis-C18（50mm×4.6mm I.D.，5μm粒径）分离后，进入MS进行检测。选用电喷雾离子化源（ESI），采用正离子多重反应监测的模式（MRM），用于定量分析的离子对分别为：m/z427.3→m/z261.0（伊潘立酮），m/z429.3→m/z261.0（代谢产物P88），m/z429.3→m/z261.0（代谢产物P95）和m/z256.3→m/z167.1（内标苯海拉明）。根据美国食品药品监督管理局（FDA）的相关规定，本研究对该方法进行了完整的确证，考察了该方法的精密度及准确度、专属性、线性范围、最低定量下限、提取回收率、基质效应及各个条件下的稳定性等。本临床试验方案主要依照《赫尔辛基宣言》、《中华人民共和国药品管理法》、等有关规定制定，选择16名健康受试者口服1mg、3mg两个剂量的伊潘立酮片后，经不同时间采集血浆样品并用LC-MS/MS方法测定血浆样品中伊潘立酮及其代谢产物P88、P95的血药浓度，绘制其药物浓度-时间曲线，利用DAS3.0（Drugand statistics）软件计算药代动力学参数，并用SPSS进行分析，研究伊潘立酮在人体内的药代动力学规律。研究结果本研究建立的人血浆样品中同时定量分析伊潘立酮及其代谢产物P88、P95的LC-MS/MS方法，在三种待测物和内标苯海拉明的出峰位置处均没有发现内源性干扰；伊潘立酮及其代谢产物P88、P95的线性范围均为5-1500pg/mL，最低定量下限均为5pg/mL，低、中、高三个浓度QC样品的准确度、日内及日间精密度均不超过±15%；伊潘立酮及其代谢产物P88、P95低、中、高三个浓度提取回收率分别为85.89±4.08%，95.17±6.93%，86.07±1.07%；97.98±5.29%，92.29±2.58%，84.04±0.34%和110.96±2.81%，97.00±7.68%，102.04±1.49%，提取回收率高并且稳定；伊潘立酮及其代谢产物P88、P95的基质效应分别为81.96±5.10%，85.55±9.09%，95.86±1.97%；88.20±2.68%，91.18±10.05%，97.51±1.75%和91.27±5.04%，84.18±3.80%，98.39±2.02%，，基质影响很小；血浆样品在未经提取处理和提取处理后的多种条件下稳定性良好，相对误差均不超过±15%；血浆样品经稀释后10倍后再提取的准确度和精密度均不超过±15%，血浆样品的稀释不影响样品的测定。本课题所建立的人血浆样品中同时定量分析伊潘立酮及其代谢产物P88、P95的LC-MS/MS方法，灵敏度高、选择性好、分析时间短、重现性好且精密准确，为伊潘立酮的临床药代动力学研究提供了支持。 16名中国健康受试者口服给予1mg和3mg伊潘立酮片，利用建立的LC-MS/MS方法检测其血浆药物浓度，绘制伊潘立酮及代谢产物P88和P95的药物浓度-时间曲线，计算其药代动力学参数。对伊潘立酮及代谢产物P88和P95的主要药代动力学参数（AUC0-t、AUC0-∞和Cmax）进行线性相关性分析。结果显示，单次口服1~3mg剂量范围内的伊潘立酮片后，伊潘立酮在人体内的药代动力学过程符合非线性动力学过程。代谢产物P88和P95的药代动力学过程是否符合线性药动学过程暂无法判断。
[Abstract]:research objective
To establish a quantitative analysis of ILP and its metabolites P88 and P95 by liquid chromatography tandem mass spectrometry (LC-MS/MS) in human plasma samples, which meet the clinical pharmacokinetic requirements of ILP (ILP), and determine the concentration of ILP and its metabolites P88, P95 drug concentration in the plasma samples of Chinese healthy volunteers, and draw the concentration of three kinds of drugs to be measured. The time curve was used to calculate its pharmacokinetic parameters, and to investigate its pharmacokinetic characteristics in vivo, so as to provide data support for the listing of ilpridone tablets.
Using acetonitrile as a eluant solid phase extraction method, the LC-MS/MS analysis of peridone and its metabolites P88, P95 in human plasma was carried out with the gradient elution of methanol -10mM acetate -0.1% formic acid solution as the mobile phase, and the separation of three kinds of samples and the internal standard SUPELCOAscentis-C18 (50mm x 4.6mm I.D., 5 micron m particle size) of the internal standard was separated. In MS, the electrospray ionization source (ESI) was selected and the mode of positive ion multiple reaction monitoring (MRM) was used. The ion pairs used for quantitative analysis were m/z427.3 to m/z261.0 (Pan Litong), m/z429.3 to m/z261.0 (metabolite P88), m/z429.3 to m/z261.0 (P95 of metabolites) and m/z256.3 to (internal standard phenhydramine). The relevant provisions of the food and Drug Administration (FDA) of the United States were confirmed in this study. The precision and accuracy of the method, specificity, linear range, minimum quantitative lower limit, extraction recovery, matrix effect and stability under various conditions were investigated.
The clinical trial scheme was formulated in accordance with the Helsinki declaration, the drug management law of People's Republic of China, and other relevant regulations. 16 healthy subjects were chosen to take oral 1mg, 3mg and two doses of peridone tablets. The plasma samples were collected at different time and LC-MS/MS method was used to determine the plasma samples and their metabolites P88, P95 The drug concentration time curve was plotted and the pharmacokinetic parameters were calculated by DAS3.0 (Drugand Statistics) software. The pharmacokinetics of peridone in human body was studied by SPSS analysis.
In the human plasma samples established in this study, the quantitative analysis of peridone and its metabolite P88, P95 LC-MS/MS method, no endogenous interference was found at the peak position of three kinds of subjects and internal standard diphenhydramine; the linear peri of irido Pan Litong and its metabolite P88, P95 were 5-1500pg/mL, and the lowest quantitative lower limit was 5pg/mL The accuracy of the QC samples of low, middle and high concentrations was not more than 15%. The recovery rates of the extraction of Pan Litong and its metabolites P88, P95, middle and high levels were 85.89 + 4.08%, 95.17 + 6.93%, 86.07 + 1.07%, 97.98 + 5.29%, 92.29 +, 92.29 +, 84.04 + 6.93%. The yield was high and stable, and the matrix effects of peridone and its metabolites P88, P95 were 81.96 + 5.10%, 85.55 + 9.09%, 95.86 + 1.97%, 88.20 + 2.68%, 91.18 + 10.05%, 97.51 + 1.75% and 91.27 + 5. The stability is good, the relative error is not more than 15%. The accuracy and precision of the plasma samples are not more than 15% after 10 times the dilution, and the dilution of the plasma samples does not affect the determination of the samples. The human plasma samples set up in this subject are simultaneously quantifying the LC-MS/MS method of the Irone and its metabolite P88, P95, and the spirit of the plasma sample. High sensitivity, good selectivity, short analysis time, good reproducibility and accuracy, which provide support for the clinical pharmacokinetics of irperidone.
16 Chinese healthy subjects were given 1mg and 3mg eperidone tablets. The concentration of plasma drugs was detected by the established LC-MS/MS method. The drug concentration time curves of eperidone and metabolic products P88 and P95 were plotted and the pharmacokinetic parameters were calculated. The main pharmacokinetic parameters of peridone and metabolic products P88 and P95 (AUC0-t, AUC0- infinity and Cmax) were used for linear correlation analysis. The results showed that the pharmacokinetic process of peridone in the human body was in line with the nonlinear kinetic process after a single dose of 1~3mg dose within a single dose of 1~3mg. The pharmacokinetic process of the metabolites P88 and P95 was not judged by the linear pharmacokinetics process.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R969.1

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