沙格雷酯药物代谢动力学研究

发布时间：2017-12-27 12:03

本文关键词：沙格雷酯药物代谢动力学研究　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究目的沙格雷酯为抗血栓形成药,主要用于改善慢性动脉闭塞症引起的溃疡、疼痛以及冷感等缺血性症状,其出血并发症少,在治疗糖尿病病人血管病变上比阿司匹林更有效。沙格雷酯,化学名称为(±)-2-(二甲胺基)-1-[[2-(3-甲氧基苯乙基)苯氧基]甲基]乙基丁二酸单酯,属于酯类药物,而酯类药物易被血浆中的酯酶水解,并且由于自身结构原因,沙格雷酯在见光的条件下不稳定,因此测定血浆中沙格雷酯含量面临着一定的挑战。本论文在加入NaF(酯酶抑制剂)、避光保持沙格雷酯结构稳定的前提下,分别研究饮食与不同制剂对沙格雷酯在人体内的药代动力学行为的影响,本研究采用了多重反应监控(MRM)的LC-MS/MS技术,建立了适用于检测人血浆中沙格雷酯的分析方法,将此方法用于测定人血浆中沙格雷酯的血药浓度,计算沙格雷酯人体内药物代谢动力学相关参数,并对其药物代谢动力学参数进行比较分析,为沙格雷酯在中国人体内生物等效性的研究提供科学依据。研究方案建立了适用于检测人血浆中沙格雷酯的LC-MS/MS分析方法。人体生物样品经有机试剂沉淀并利用固相萃取技术处理去除杂质后,取20μl上清液,经过Agela Technologies Venusil MP C18(4.6×100 mm,5μm)色谱柱,通过不同梯度的有机相和水相洗脱的方式进行液相色谱分离,流动相A为含0.02%甲酸的10 mM醋酸铵溶液,流动相B为乙腈,LC-MS/MS系统用时为4.5 min。MS/MS系统采用ESI电喷雾离子源,正离子多重反应离子扫描模式(MRM),用于定量的离子对分别为:沙格雷酯(430.3/135.1,m/z),内标地西泮(285.2/193.2,m/z)。本液相色谱-串联质谱的测定方法依照国家相关指导原则的要求,分别从特异性SPE、回归线性、最低定量下限(LLOQ)、稀释方法学(dilute)、准确度、精密度、基质效应(ME)、提取回收率(RE)、残留效应(carry over)及所涉及的稳定性等方面对本论文的LC-MS/MS方法进行方法确证。利用所建立的LC-MS/MS定量分析方法测定人血浆中沙格雷酯的血药浓度,并且绘制了沙格雷酯药物浓度-时间曲线(即药时曲线)。根据沙格雷酯的药时曲线得出沙格雷酯药物代谢动力学的相关参数(药时曲线下面积AUC0-∞、达峰时间Tmax、达峰浓度Cmax、消除半衰期t1/2等),并对以上药代动力学参数进行分析,用于沙格雷酯药代动力学过程进行相关评价。研究结果本论文建立了基于多重反应监测(MRM)模式检测人血浆中沙格雷酯的液相色谱-串联质谱(LC-MS/MS)的分析方法,本方法有高度的灵敏度并且分析速度快,沙格雷酯和内标地西泮的批内和批间的准确度和精密度均达到了测试要求;经过沉淀蛋白和固相萃取的两部分样品处理后,沙格雷酯和地西泮基本不受其他干扰性物质的影响,同时具有较高的提取回收率;本方法能够使待测物沙格雷酯的稳定性在采血、运输、存储和分析过程中得到保证。本方法符合CFDA指导原则的要求,满足沙格雷酯药代动力学研究的要求,适用于沙格雷酯药代动力学及生物等效性的研究。
[Abstract]:The purpose of the study is sarpogrelate antithrombotic drugs, mainly used to improve chronic arterial occlusive disease caused by the ulcer, pain and cold ischemic symptoms, the fewer bleeding complications, more effective than aspirin in the treatment of diabetic vascular disease. Sarpogrelate, the chemical name is (+) -2- (two -1-[[2- (methylamino) 3- methoxy phenyl) phenoxy] methyl] ethyl succinate ester, belonging to drugs, and drug susceptible esterase hydrolysis esters in plasma, and because of their structure, Shug Ray did not see the light in the stable under the condition, so the determination of sarpogrelate hydrochloride in plasma facing certain challenges. In this paper, adding NaF (esterase inhibitor), light of sarpogrelate stable structure, respectively to study the effect of diet and different preparation of sarpogrelate hydrochloride in human body pharmacokinetics, this study adopts multiple reaction monitoring (MRM) of the LC-MS/MS technology, analysis method was established for detection of sarpogrelate in human plasma, the method for determination of plasma concentration in human plasma by Shug Ray, the calculation of sarpogrelate body pharmacokinetic parameters, and the pharmacokinetic parameters were compared and analyzed, as in China sarpogrelate in human bioequivalence study to provide scientific basis for. The research program was established for detection of sarpogrelate hydrochloride in human plasma by LC-MS/MS method. Human biological samples by organic reagents by solid phase extraction technique and precipitation treatment to remove impurities, take 20 l after Agela Technologies Venusil MP by C18 (4.6 * 100 mm, 5 m) column, through the organic and aqueous phases of different gradient elution methods of liquid chromatography separation, mobile phase containing A 0.02% formic acid 10 mM ammonium acetate solution, the B of mobile phase acetonitrile, LC-MS/MS system is 4.5 min. The MS/MS system adopts ESI electrospray ion source and positive ion multiplex reactive ion scanning mode (MRM). It is used for quantitative ion pair, namely Sagre ester (430.3/135.1, m/z), internal standard diazepam (285.2/193.2, m/z). The liquid chromatography tandem mass spectrometry method for the determination of the related guidelines in accordance with national requirements, from the specific SPE, linear regression, the lowest limit of quantification (LLOQ) and dilution methodology (dilute), accuracy, precision, matrix effect (ME) and the recovery rate (RE), residual effect (carry over) and the stability and other aspects of the LC-MS/MS method confirmed this method. Determination of serum concentration of sarpogrelate hydrochloride in human plasma by LC-MS/MS quantitative analysis method is established, and draw the sarpogrelate drug concentration time curve (i.e. the concentration time curve). According to the relevant parameters of sarpogrelate pharmacokinetic curve that the pharmacokinetics of the drug ester (area under the concentration time curve of AUC0- ~ Tmax, peak time, peak concentrations of Cmax, t1/2, and the elimination half-life) on the pharmacokinetic parameters were analyzed for evaluation of sarpogrelate pharmacokinetics. The research results of this thesis established a multiple reaction monitoring (MRM) mode based on the detection of human plasma sarpogrelate liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis method, this method has high sensitivity and fast analysis, Shug Ray and the internal standard of diazepam in batch and inter batch accuracy and precision were achieved the test requirements; after protein precipitation and solid phase extraction of two samples after treatment, sarpogrelate and diazepam are not affected by the interference of other substances, but also has high recovery rate; stability of this method can test Shug Ray to be guaranteed in the blood collection, transportation, storage and analysis process. This method meets the requirements of CFDA guidelines, meet the kinetics of sarpogrelate pharmacokinetics requirements, applicable to the study of sarpogrelate pharmacokinetics and bioequivalence.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96

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