氨肽酶N抑制剂在大鼠体内的药动学研究

发布时间：2018-07-08 17:48

  本文选题：氨肽酶N抑制剂 + Bes-5FU　； 参考：《青岛大学》2014年硕士论文

【摘要】：目的 筛选出抗肿瘤活性较高的氨肽酶N抑制剂,建立该化合物体内、体外的含量检测方法并对其在大鼠体内的药物代谢动力学进行研究。 方法 1、分别采用高效液相色谱法、MTT比色法等方法对候选化合物的稳定性、药理活性进行评价、筛选。 2、采用蛋白沉淀的血浆预处理方法处理Bes-5FU的血浆样品,色谱柱为Venusil ASB C18(4.6mm×250mm,5μm)柱,Bes-5FU的流动相为乙腈-0.05%甲酸(22：78,v/v),流速为1ml/min,紫外检测波长为262.7nm,并对方法的线性、灵敏度、精密度、提取率、稳定性等进行考察。 3、采用液液萃取的血浆预处理方法处理5-氟尿嘧啶的血浆样品,色谱柱为Venusil ASB C18(4.6mm×250mm,5μm)柱,5-氟尿嘧啶的流动相为甲醇-水(5：95,v/v),流速为1ml/min,紫外检测波长为266.3nm,并对方法的线性、灵敏度、精密度、提取率、稳定性等进行考察。 4、进行大鼠体内药物代谢动力学研究,健康Wistar雄性大鼠六只,体重250-260g,实验前禁食12h,但自由饮水,于给药后0.033,0.083,0.167,0.333,0.5,1,2,3,4,6,8和10h经眼底静脉丛取血后置于经肝素抗凝的EP管中,于4℃、8609×g离心3min分离血浆,分别采用蛋白沉淀法和液液萃取法进行血浆样品的预处理；采用高效液相色谱法进行血浆样品的检测,记录峰面积,绘制药时曲线；采用DAS2.0分析软件(中国药理学会),利用非隔室模型计算给药后的药动学参数。 结果 1、筛选出的目标化合物Bes-5FU在药理活性、稳定性等方面均符合要求。 2、Bes-5FU的保留时间为11.5min；标准曲线为：y=5336.1x-1702.1(r=0.9996),最低定量限为：0.3μg/ml; Bes-5FU低、中、高(0.5、5和10μg/m1)三个质量浓度的提取回收率分别为65.3±5.2%,66.7±4.3%和68.2±4.1%(n=6)；Bes-5FU的低、中、高三个质量浓度(0.5、5和10μg/m1)的日内、日间精密度RSD均小于15%(n=6)。 3、5-氟尿嘧啶的保留时间为6min；标准曲线为：y=37118x-9030.1(r=0.9998),最低定量限为：0.1μg/ml；5-氟尿嘧啶低、中、高(0.25、5和10μg/ml)三个质量浓度的提取回收率分别为70.4±7.4%,76.2±7.1%和75.7±6.8%(n=6)；5氟尿嘧啶的低、中、高三个质量浓度(0.25、5和10μg/ml)的日内、日间精密度RSD均小于10.0%(n=6)。 4、采用大鼠股静脉注射给药,给药剂量为84mg/kg,给药后血浆中Bes-5FU的主要代谢产物5-氟尿嘧啶的主要药动学参数t1/2(h)、MRT0-t(h)、AUC0-t(μg·h/mL)、 AUC0-∞(μg·h/mL)、AUMC0-t(μg·h2/mL)、AUMC0-∞(μg·h2/mL)分别为0.14±0.04、0.17±0.04、17.73±6.27、17.78±6.31、2.77±0.59、3.07±0.63。 结论 1、筛选出的目标化合物Bes-5FU稳定性较好、药理活性较高,可对其成药性进行深入研究。 2、建立了Bes-5FU的高效液相检测方法,该方法灵敏度高、准确性好,可用于Bes-5FU在大鼠体内的药物代谢动力学研究。 3、对Bes-5FU在大鼠体内药代动力学特征进行了初步评价,其作为前药在大鼠体内较快地代谢成5-氟尿嘧啶而发挥抗肿瘤作用。
[Abstract]:Objective to screen the aminopeptidase N inhibitor with high antitumor activity, to establish a method for the determination of its content in vivo and in vitro and to study its pharmacokinetics in rats. Methods 1. The stability and pharmacological activity of the candidate compounds were evaluated and screened by HPLC and MTT colorimetry respectively. 2. The plasma samples of Bes-5FU were treated by the plasma pretreatment method of protein precipitation. The mobile phase of Bes-5FU was acetonitrile-0.05% formic acid (22: 78 v / v), the flow rate was 1 ml / min, the UV detection wavelength was 262.7 nm, and the linearity, sensitivity, precision, extraction rate of the method were obtained. The chromatographic column was Venusil ASB C18 (4.6mm 脳 250mm-1 5 渭 m) and the mobile phase was acetonitrile-0.05% formic acid (22: 78 v / v), the flow rate was 1 ml / min and the UV detection wavelength was 262.7 nm. (3) the plasma samples of 5-fluorouracil were treated by liquid-liquid extraction plasma pretreatment method. The chromatographic column was Venusil ASB C18 (4.6mm 脳 250mm-1 5 渭 m) column. The mobile phase of 5-fluorouracil was methanol-water (5: 95 v / v), the flow rate was 1 ml / min, and the UV detection wavelength was 266.3 nm. (4) the pharmacokinetics of rats was studied. Six healthy male Wistar rats, weighing 250-260 g, fasted for 12 hours before the experiment, but drank freely. The blood samples were collected from the fundus venous plexus of eyes for 10 hours and then placed in EP tubes with heparin anticoagulant. The plasma was separated by centrifugation 3min at 4 鈩，

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