【作者】 王楠；

【导师】 朱立勤；

【作者基本信息】 天津医科大学 ， 药理学， 2012， 硕士

【摘要】 目的:建立感染患者中左氧氟沙星、莫西沙星的群体药动学模型,并研究药动/药效相关性,为优化左氧氟沙星、莫西沙星抗感染治疗方案提供依据。方法:1.建立氟喹诺酮类药物血浆浓度测定方法:①左氧氟沙星:色谱柱:Kromasil C18(150mm×4.6mm.5μm),流动相:0.05mol/L 磷酸二氢钾(磷酸调 pH 至 3.1)-乙腈(85:15,V:V),紫外检测波长:294nm,流速:1.OmL·min-1,进样量:10μL,柱温:室温,内标物:甲硝唑。②莫西沙星:色谱柱:Kromasil C18(250mm×4.6mm.5μm),流动相:1%三乙胺(磷酸调 pH 至 4.8)-乙腈(80:20,V:V),紫外检测波长为296nm,流速为1.0 mL·min-1,进样量:20μL,柱温:室温,内标物:环丙沙星。2.左氧氟沙星群体药动/药效学模型研究:采集27名成年感染疾病患者,左氧氟沙星500mg每日—次静脉滴注的血浆样本49份。用药过程中取血2次,取血时间分别为静脉滴注后立即取样,第3-5天给药前取样,测定血药浓度。采用非线性混合效应模型(NONMEM),建立左氧氟沙星一室群体药动学模型.并采用Bootstrap进行模型验证。评价所有患者抗感染效果,采用Logistic回归的方法进行药动/药效相关性研究。3.莫西沙星群体药动/药效学模型研究:采集37名成年感染疾病患者,莫西沙星400mg每日一次静脉滴注的血浆样本67份。取样点、模型建立方法同左氧氟沙星。结果:1.建立氟喹诺酮类药物血浆浓度测定方法,①左氧氟沙星:左氧氟沙星和内标甲硝唑保留时间分别为8.3min和4.2min,最低检测浓度为0.05μg·mL-1;标准曲线为Y=2.824X+0.065(r2=0.999),检测浓度在0.05-10μg·mL-1之间线性关系良好,低、中、高三种浓度(0.1、0.5、2.0μg·mL-1)的绝对回收率分别为96.59%、93.87%、85.58%.方法回收率分别为 101.00%、97.35%、103.35%,日内 RSD 值分别为 8.73%、2.35%、0.20%.日间 RSD 值分别为 7.53%、2.73%、0.34%。②莫西沙星:莫西沙星和内标环丙沙星保留时间分别为15min和5.5min,标准曲线为 Y=3.0506X-0.0235(r2=0.9998),检测浓度在 0.025-5.0μg·mL-1 范围内线性关系良好;低、中、高三种浓度(0.05、0.5、2.5μg·mL-1)的绝对回收率分别为69.88%、78.86%、78.51%,方法回收率分别为 98.50%、96.61%、101.79%,日内 RSD 值分别为 4.12%、3.12%、1.85%,日间 RSD 值分别为 4.70%、1.16%、3.38%;重复性RSD值为1.67%。2.建立左氧氟沙星群体药动学模型,考察固定效应,AGE,AST对CL有影响(P<0.05),AGE,AST对V有影响(P<0.05),被保留在全量回归模型中。最终群体药动学模型为:CL=8.8-2.96*(AGE/50),V=62.8-0.33*AGE+0.302*AST。内部验证300次bootstrap中,273次成功运行,成功率为91.0%,获得的参数均值与最终模型参数比较差异无显著性。左氧氟沙星药动/药效相关性模型为P(y=EFF)= Exp(-4.607+0.87AUC/MIC)/(1+Exp(-4.607+0.87AUC/MIC))。3.建立莫西沙星群体药动学模型,考察固定效应,WT,AGE对CL有影响(P<0.05),WT对V有影响(P<0.05),被保留在全量回归模型中。最终群体药动学为:CL=16.8-4.55*(AGE/50),V=90.2+0.78]*(WT-70)。内部验证结果显示.400次bootstrap中,382次成功运行,成功率为95.5%.获得的参数均值与最终模型参数比较差异无显著性。莫西沙星药动/药效相关性模型为P(y=EFF)=Exp(4.085+0.051AUC/MIC-0.097AGE)/(1+Exp(4.085+0.051AUC/MIC-0.097AGE))结论:研究中建立的测定方法操作简便、分析快速、准确、灵敏度高、亏属性和重现性好,可以满足临床血药浓度监测和药动学研究的要求。左氧氟沙星、莫西沙星群体药动学模型预测值与实测值相关性良好,标本采集时间和预测浓度与权重残差间无趋势性分布,模型稳定可靠,可用于模拟个体药动学参数。利用群体数据建立的药动/药效相关性模型可为临床个体化给药提供依据。更多还原

【Abstract】 OBJECTIVE:To establish the population pharmacokinetic model and explorer PK/PD relationship of levofloxacin and moxifloxacin in patients with infectious disease,so as to provide basis for the optimal treatment regimen.METHODS:1.①The determination of levofloxacin sample was performed on Kromasil C18(150mm×4.6mm.5μm)column with a fixed sample injection volume of 10μL.The mobile phase was 0.05mol·L-1 potassium dihvdrogen phosphate solution(pH adjusted to 3.1 by phosphoric acid)-acetonitrile(85:15,V:V)at a flow rate of 1.0 mL·min-1.The UV detective wavelength was set at 294nm;the column temperature was the same as the room temperature and metronidazole was chosen as the internal standard.②The determination of moxifloxacin sample was performed on Kromasil C18(250mm×4.6mm.5μm column with a fixed sample injection volume of 20μL.The mobile phase was 1%triethviamine(pH adjusted to 4.8 by phosphoric acid)-acetonitrile(80:20,V:V)at a flow rate of 1.0 mL·min-1.The UV detective wavelength was set at 296nm the column temperature was the same as the room temperature and ciprofloxacin was chosen as the internal standard.2.PPK/PD model of levofloxacin:A population pharmacokinetic model of levofloxacin was constructed from data of adult patients with infectious disease.A total of 27 patients treated with intravenous levofloxacin 500mg every day.Totally 49 plasma samples of levofloxacin were collected either immediately after intravenous dripping or before application on the 3rd.4th or 5th day.Nonliner mixed effect model was employed using a one-compartment model of levofloxacin.Bootstrap was applied to validate the model.Evaluating the anti-infective effects.and study the PK/PD relationship by using logistic regression.3.PPK/PD model of moxifloxacin:A population pharmacokinetic model of moxifloxacin was constructed from data of adult patients with infectious disease.A total of 37 patients treated with intravenous moxifloxacin 400mg every day.Totally 67 plasma samples of moxifloxacin were collected.Other steps were the same as levofloxacin.RESULTS:1.①Levofloxacin:the retention time of metronidazole and levofloxacin were 4.2min and 8.3min respectively.The lowest detectable concentration was O.Sμg.mL-1.The calibration curve was Y=2.824X+0.065(r2=0.999)and it was linear within the concentration range of 0.05-10μg·mL-1.The absolutely recoveries of moxifloxacin at three concentrations(0.1,0.5,2.0μg·mL-1)were 96.59%.93.87%.85.58%,respectively;the method recoveries were 101.00%.97.35%,103.35%.respectively;with inter-day RSD at 8.73%,2.35%.0.20%and intra-day RSD at 7.53%,2.73%.0.34%.respectively.②Moxifloxacin:The retention time of moxifloxacin and ciprofloxacin were about 15 min and 5.5min respectively’.The calibration curve was Y=3.0506X-0.0235(r2=0.9998)and it was linear within the concentration range of 0.025-5.0μg·mL-1.The absolutely recoveries of moxifloxacin at three concentrations(0.05、0.5、2.5μg·mL-1)were 69.88%.78.86%.78.51%.respectivel)y;the method recoveries were 98.50%,96.61%,101.79%,respectively;with inter-day RSD at 4.120%,3.12%,1.85%and intra-day RSD at 4.70%.1.16%,3.38%,respectively.And the repeatability RSD was 1.67%.2.PPK model of levofloxacin was established.Validating the fixed effect.AGE.AST were identified as intrinsic factors which sigynificantly affected clearance(P<0.05)and AGE.AST significantly affected the apparent volume of distribution(P<0.05).The final model was CL=8.8-2.96*(AGE/50),V=62.8-0.33*AGE+O.302*AST.Internal validation for 300 times in the bootstrap.273 runs successfully.The success rate was 91.0%.The difference between parameter value and final model parameters was not significant.The PK/PD relationship was P(y=EFF)=Exp(-4.607+0.87AUC/MIC)/(1+Exp(-4.607+0.87AUC/MIC)).3.PPK model of levofloxacin was established.Validating fixed effect.WT.AGE were identified as intrinsic factors which significantly affected clearance(P<0.05)and WT significantly affected the apparent volume of distribution(P<0.05).The final model was CL=16.8-4.55*(AGE/50),V=90.2+0.781*(WT-70).Internal validation for 400 times in the bootstrap.382 runs successfully.The success rate was 95.5%.The difference between parameter value and final model parameters was not significant.The PK/PD relationship was P(y=EFF)=Exp(4.085+0.051AUC/MIC-0.097AGE)/(1+Exp(4.085+0.051AUC/MIC-0.097AGE)).CONCLUSION:The HPLC method is simple,rapid,accurate,and had high sensitivity,good specificity and reproducibility,which can be used to determine the drug concentration in plasma for clinical and pharmacokinetic research.Good correlation was showed between forecast values and measured values in both levofloxacin and moxifloxacin PPK models.There were no trends of distribution between weighted residuals and sample collection time,prediction of concentration.The models were stable and reliable that can be used to simulate individual pharmacokinetic parameters.The PK/PD relationship can provide basis for individual medication.更多还原