本文選題：氨肽酶N抑制劑 + Bes-5FU　； 參考：《青島大學(xué)》2014年碩士論文

【摘要】：目的 篩選出抗腫瘤活性較高的氨肽酶N抑制劑,建立該化合物體內(nèi)、體外的含量檢測方法并對(duì)其在大鼠體內(nèi)的藥物代謝動(dòng)力學(xué)進(jìn)行研究。 方法 1、分別采用高效液相色譜法、MTT比色法等方法對(duì)候選化合物的穩(wěn)定性、藥理活性進(jìn)行評(píng)價(jià)、篩選。 2、采用蛋白沉淀的血漿預(yù)處理方法處理Bes-5FU的血漿樣品,色譜柱為Venusil ASB C18(4.6mm×250mm,5μm)柱,Bes-5FU的流動(dòng)相為乙腈-0.05%甲酸(22：78,v/v),流速為1ml/min,紫外檢測波長為262.7nm,并對(duì)方法的線性、靈敏度、精密度、提取率、穩(wěn)定性等進(jìn)行考察。 3、采用液液萃取的血漿預(yù)處理方法處理5-氟尿嘧啶的血漿樣品,色譜柱為Venusil ASB C18(4.6mm×250mm,5μm)柱,5-氟尿嘧啶的流動(dòng)相為甲醇-水(5：95,v/v),流速為1ml/min,紫外檢測波長為266.3nm,并對(duì)方法的線性、靈敏度、精密度、提取率、穩(wěn)定性等進(jìn)行考察。 4、進(jìn)行大鼠體內(nèi)藥物代謝動(dòng)力學(xué)研究,健康Wistar雄性大鼠六只,體重250-260g,實(shí)驗(yàn)前禁食12h,但自由飲水,于給藥后0.033,0.083,0.167,0.333,0.5,1,2,3,4,6,8和10h經(jīng)眼底靜脈叢取血后置于經(jīng)肝素抗凝的EP管中,于4℃、8609×g離心3min分離血漿,分別采用蛋白沉淀法和液液萃取法進(jìn)行血漿樣品的預(yù)處理；采用高效液相色譜法進(jìn)行血漿樣品的檢測,記錄峰面積,繪制藥時(shí)曲線；采用DAS2.0分析軟件(中國藥理學(xué)會(huì)),利用非隔室模型計(jì)算給藥后的藥動(dòng)學(xué)參數(shù)。 結(jié)果 1、篩選出的目標(biāo)化合物Bes-5FU在藥理活性、穩(wěn)定性等方面均符合要求。 2、Bes-5FU的保留時(shí)間為11.5min；標(biāo)準(zhǔn)曲線為：y=5336.1x-1702.1(r=0.9996),最低定量限為：0.3μg/ml; Bes-5FU低、中、高(0.5、5和10μg/m1)三個(gè)質(zhì)量濃度的提取回收率分別為65.3±5.2%,66.7±4.3%和68.2±4.1%(n=6)；Bes-5FU的低、中、高三個(gè)質(zhì)量濃度(0.5、5和10μg/m1)的日內(nèi)、日間精密度RSD均小于15%(n=6)。 3、5-氟尿嘧啶的保留時(shí)間為6min；標(biāo)準(zhǔn)曲線為：y=37118x-9030.1(r=0.9998),最低定量限為：0.1μg/ml；5-氟尿嘧啶低、中、高(0.25、5和10μg/ml)三個(gè)質(zhì)量濃度的提取回收率分別為70.4±7.4%,76.2±7.1%和75.7±6.8%(n=6)；5氟尿嘧啶的低、中、高三個(gè)質(zhì)量濃度(0.25、5和10μg/ml)的日內(nèi)、日間精密度RSD均小于10.0%(n=6)。 4、采用大鼠股靜脈注射給藥,給藥劑量為84mg/kg,給藥后血漿中Bes-5FU的主要代謝產(chǎn)物5-氟尿嘧啶的主要藥動(dòng)學(xué)參數(shù)t1/2(h)、MRT0-t(h)、AUC0-t(μg·h/mL)、 AUC0-∞(μg·h/mL)、AUMC0-t(μg·h2/mL)、AUMC0-∞(μg·h2/mL)分別為0.14±0.04、0.17±0.04、17.73±6.27、17.78±6.31、2.77±0.59、3.07±0.63。 結(jié)論 1、篩選出的目標(biāo)化合物Bes-5FU穩(wěn)定性較好、藥理活性較高,可對(duì)其成藥性進(jìn)行深入研究。 2、建立了Bes-5FU的高效液相檢測方法,該方法靈敏度高、準(zhǔn)確性好,可用于Bes-5FU在大鼠體內(nèi)的藥物代謝動(dòng)力學(xué)研究。 3、對(duì)Bes-5FU在大鼠體內(nèi)藥代動(dòng)力學(xué)特征進(jìn)行了初步評(píng)價(jià),其作為前藥在大鼠體內(nèi)較快地代謝成5-氟尿嘧啶而發(fā)揮抗腫瘤作用。
[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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