连用OK-432后对5-氟尿嘧啶在小鼠体内药代动力学的影响

【作者】 刘新华；

【导师】 詹丽芬；

【作者基本信息】 中国医科大学 ， 药理学， 2003， 硕士

【摘要】 目的 用RP-HPLC法检测5-氟尿嘧啶在小鼠体内的血药浓度，研究连用OK-432后对5-氟尿嘧啶在小鼠体内药代动力学的影响，为临床合理用药提供实验依据。 方法 血浆样品的处理及测定：取血浆200μl于1.0ml EP管中，精密加入200μg·ml^-1的内标5-Bru溶液25μl和10％高氯酸200μl，涡旋振荡1min，混匀，离心5min(10900r·min^-1)，吸取上清液过滤（0.45μm微孔滤过膜），取10μl进样测定，重复三次取其均值。 给药方法及血样采集：单用5-Fu组分别腹腔注射5-Fu生理盐溶液75.8mg·kg^-1(浓度为7.58mg·ml^-1，0.2ml/20g)于给药前和给药后1、3、5、10、20、30、60、90、120分钟分别断头采血于肝素化的离心管中，冰箱保存，直至采血结束后，全部样品离心15min(3000r·min^-1)，取上清液200μl于EP管中，置-20℃冰箱保存待测。另一组小鼠连续2日（每日2次）腹腔注射OK-432生理盐溶液25KE·kg^-1(浓度为2.5KE·ml^-1，0.2ml/20g，临用时配制)。于第3日给OK-432后30min按75.8mg·kg^-1用量腹腔注射5-Fu，采血时间及方法同上。 结果 1．血浆中5-Fu测定方法的认证 5-Fu色谱行为及专一性：5-Fu的保留时间约为4.0min，内标的保留时间约为7.1min，在本实验条件下5-Fu和5-Bru均有较大的吸收峰并有较好的分离度，血浆内源性杂质不干扰对5－Fu的含量测定。 血浆样品标准曲线：于空白血浆中分别加人5－Fu储备溶液，使其浓度为 10050在5＊0＊刀．lug·rnl－’的系列血浆标准液，按血浆样品处理方法操作，以样品的色谱吸收峰面积（AS）与内标的色谱吸收峰面积（Al）比值门)对样品浓度＊)作直线回归，标准曲线方程为 Y＝0．059 C－0．0040（r＝0．9993）。血浆浓度在 0．二一100pg·IIl－‘范围内线形关系良好，按条件测得5－Fu最低检测限0·0卜g·Inl－l 血浆样品回收率与精密度实验：分别用空白血浆配制5－Fu浓度为50在5、10、l．opg·Inl－’的样品，按血浆样品的测定方法操作，计算方法的回收率＼日内及日间变异。结果表明，在高中低三个浓度下方法回收率为 93．99－103．26％，日内及日间变异均小于10％。 2。单用 5－Fu与连用 OK432后的 5－Fu药代动力学研究 血药浓度：将两组各时间点的血样按血浆样品测定方法操作后，计算出相应的血药浓度。血浆浓度呈快。相，慢p相衰减。 药代动力学参数：将药时数据经“3 P97”程序处理，结果表明，连用OK斗32后使5FU的吸收减慢，半衰期延长，达峰时间延缓，峰浓度下降，血药时曲线下面积增加，消除明显减慢。 结 论 连用0＊＊32后，5I 的药时曲线下面积＊＊C增大，进人血循环总药量增加，消除速率半衰期T；加增加，作用时间延长，为提高疗效减少毒副作用，两药联用时注意调整剂量和给药间隔。更多还原

【Abstract】 ObjectiveThe concentrations of 5-fluorouracil in the blood of mice were detected by RP - HPLC, the effects of OK-432 on the pharmacokinetics of 5-fluorouracil were studied when the former was followed by the latter, which provided basis for clinic.Method1. Disposal and determination of blood samples200 l of plasma were injected in 1.0 ml EP blanket , 25 l of 200 g ’ml-1 internal solutions 5-bromouracil and 200 l of 10% perchloric were added. The mixture were vortexed for 1 minute and then centriFuged at 10900 r min-1 for 5 min at room temperature. The supernatant was filtrated by filter membrane with 0.45 m micropore. 10 l of the filtrated solution was injected into HPLC.2. Administration of drug and collection of blood samples 5-fluorouracil of 75. 8 mg kg-1 (the concentration is 7.58 mg ml-1,0.2 ml/20 g) was injected into the abdominal cavity of mice in 5-fluoroural group. Blood samples were collected in heparinized cen-triFuge tube before and 1, 3 , 5, 10, 20, 30, 60, 90 and 120 minutes after the injection of 5-fluorouracil. All blood samples were centri-fuged for 15 minutes at 3000 r min-1. 200 l of supernatant was immediately added in EP blanket and stored in - 20 . OK-432 of 25KE kg -1 ( concentration is 2.5KE -ml-1 ,0.2 ml/20g)was injected into the abdominal cavity of mice in another group, the mice was given 5-fluorouracil after 30 minute of injection of OK-432 on the thirdday. The time and method of samplings were the same as mentioned a-bove.3. Statistical analysisThe results were analyzed with the 3p97 program designed by the Chinese Pharmacological Society to determine the compartment models and the pharmacokinetics parameters.Results1. Identification of determination of 5 -fluorouracil in plasma1. 1 Chromatograms of 5-fluorouracil and the specificityThe retention time of 5-fluorouracil is about 4. 0 min and retention time of internal standard is about 7. 1 min. Plasma endogenous materials dont interfere with the content of 5-fluorouracil.1.2 Standard curve5-Fu of various known concentration was added to plasma at the final concentration of 100, 50, 25, 10, 1 and 0. 1 g ml-1. They were processed according to the above procedure. The linear regression was constructed by using peak area comparison of the 5-Fu and 5-Bru and corresponding concentration. Satisfactory linearity was observed in the range of 0.1 - 100 g ml-1 of the drug. The regression equation was Y =0.0591C -0. 0040 ( r =0. 9993) , in the equation , Y is the peak area comparison , C is the concentration ( g ml -1) of the drug in plasma and r is the coefficient of correlation . The limit of detection for 5-Fu under these conditions was 0. 01 g ml-1.1. 3 Recoveries and precisionKnown amounts of 5-Fu were added to plasma at the final concentration of 50 , 25 and 0.1 g ml-1 . These samples were treated ac-cording to the above inter - day and within day repeatedly. The recovery of the compound was calculated by comparing the experimental value with the corresponding theoretical value. The mean relative recovery of 5-Fu was 93. 99 ~ 103.26% , the coefficients of variation of within - day and inter - day were both under 10%. 2. Study on Pharmacokinetic2.1 Concentration of 5-Fu in blood after igThe samples were treated and the corresponding concentrations in blood were calculated. The level of the drug declined with time in the two - compartment pattern.2.2 Pharmacokinetic parametersThe concentrations of 5-Fu and the corresponding times were processed by program 3p97. The present study showed that the absorption and maximum concentration of 5-Fu decreased, the half-life and the area under the curve increased. The elimination of 5-Fu in body slowed.ConclusionThe area below the curve of 5-Fu increased when OK -432 was used in combination, the totail amount of drug in circulation increased, the half life of elimination increased, the action time was prolonged. The dosage and interval of administration should be paid more更多还原