HPD注射液的药代动力学研究

发布时间：2018-04-28 15:51

本文选题：HPD注射液 + UPLC-MS/MS　；参考：《吉林大学》2016年博士论文

【摘要】：羟基人参二醇(Hydroxylation panoxadiol,简称HPD)是首次从西洋参茎叶总皂苷分离鉴定的一种三萜类新化合物。研究表明,手性结构修饰方法制备的HPD达到了工业化水平;抗宫颈癌的药理活性较强;毒性较低;将其制成的HPD注射液具有良好开发一类创新药物的前景,其中药代动力学是开发创新药物的重要内容之一,目前大鼠体内的药代动力学研究尚未见报道。本论文首次建立了大鼠血浆、组织及排泄物中HPD的UPLC-MS/MS定量分析方法,研究了经静脉给予大鼠HPD注射液,药物在大鼠体内的吸收、分布和排泄规律,同时测定了HPD在体外的血浆蛋白结合率;首次建立了大鼠体内HPD代谢物的UPLC-Q-TOF/MS定性分析方法,鉴定了其代谢产物,揭示了HPD在大鼠体内的代谢规律。取得以下创新成果:1、HPD注射液的血药浓度-时间曲线研究首次建立了大鼠血浆中HPD的UPLC-MS/MS定量分析方法。测定了经静脉给予大鼠低、中、高三个剂量组(8.0,16.0,32.0mg/kg)的HPD注射液后,不同时间点血浆中的药物浓度。实验数据经DAS3.0软件计算,获得的HPD注射液药代动力学的相关参数如下:低剂量组,T1/2 5.76±1.46h,Cmax5535.33±610.76μg/L,Tmax 0.10±0.00h,AUC(0-t)11670.15±1364.30μg·h/L,AUC(0-∞)12023.88±1573.57μg·h/L,Vz 5.52±1.22 L/kg,CLz 0.67±0.08 L/h/kg;中剂量组,T1/2 6.50±1.05h,Cmax 12344.29±1631.71μg/L,Tmax 0.10±0.00h,AUC(0-t)26253.13±4307.29μg·h/L,AUC(0-∞)26986.39±4635.18μg·h/L,Vz 5.63±0.90L/kg,CLz 0.61±0.10L/h/kg;高剂量组,T1/2 6.41±1.40h,Cmax 24683.57±3693.03μg/L,Tmax 0.10±0.00h,AUC(0-t)48503.57±5309.65μg·h/L,AUC(0-∞)49165.27±5335.80μg·h/L,Vz 6.07±1.41L/kg,CLz 0.66±0.07L/h/kg。结果表明,AUC、Cmax与HPD的给药剂量均成正比例关系,符合一级动力学特征;Vz 5.52-6.07L/kg提示不仅分布在血液中,在组织中也有分布;T1/2 5.76-6.50h及CLz 0.61-0.67L/h/kg提示HPD注射液的消除较慢。2、HPD注射液的分布研究首次建立了测定大鼠组织中HPD的UPLC-MS/MS定量分析方法。分别测定了大鼠静脉给予16.0mg/kg的HPD注射液后,不同时间点(15min,1h,4h,6h)大鼠13个组织(心、肺、脾、肝、肾、小肠、大肠、胃、肌肉、脂肪、脑、子宫和睾丸)中HPD的含量。结果显示,HPD快速广泛地分布到各个组织中;主要分布在肾、子宫组织中,其它各个组织中的分布情况随时间不同而各有差异;并且在各组织中均未产生蓄积作用。3、HPD注射液的血浆蛋白结合率研究首次建立了HPD在大鼠血浆、犬血浆、人血浆及透析液中的UPLC-MS/MS定量分析方法。结合平衡透析法,分别测定了HPD与上述三种血浆的蛋白结合率,结果如下:0.1μg/m L、4.0μg/m L和16μg/m L三个浓度的HPD与大鼠血浆的蛋白结合率分别为68.36%、69.07%和70.48%;与犬血浆的蛋白结合率分别为67.66%、74.42%和72.32%;与人血浆的蛋白结合率分别为65.19%、68.40%和66.09%。结果表明,在0.1μg/m L-16.0μg/m L的范围内,HPD与上述三种血浆的蛋白结合率都不具有浓度依赖性;三种血浆的蛋白结合率种属间无明显差异。4、HPD注射液的排泄研究首次建立了大鼠胆汁、尿和粪样中HPD的UPLC-MS/MS定量分析方法。分别测定了大鼠静脉给予16.0mg/kg的HPD注射液后,大鼠胆汁、尿和粪样中不同时间段HPD的含量,结果如下:胆汁在36h内累积排泄百分比仅为0.06%,而尿和粪在72h内累积排泄百分比分别为46.39%和0.18%。结果表明,HPD注射液的原型药经胆汁和尿的排泄极少,主要经粪便排泄;由于三种排泄物中总的累积排泄百分比仅为46.63%,表明其在大鼠体内的生物转化是以代谢消除为主。5、HPD注射液的代谢研究首次建立了大鼠胆汁、尿和粪样中HPD代谢物的UPLC-Q-TOF/MS定性分析方法。鉴定了大鼠经静脉给予16.0mg/kg的HPD注射液后,大鼠粪、尿和胆汁在72h内的代谢产物,结果如下:HPD注射液在大鼠体内除原形药外,共检测到21个代谢产物,其中粪样和尿样中共检测出13个代谢产物,12个为Ⅰ相代谢产物;在胆汁样品中共检测出8个代谢产物,5个为Ⅱ相代谢产物。结果表明,HPD在大鼠体内Ⅰ相代谢和Ⅱ相代谢会同时发生,而且Ⅰ相代谢产物主要发生在粪和尿中,Ⅱ相代物主要发生在胆汁中。
[Abstract]:Hydroxylation panoxadiol (HPD) is a new three terpenoid compound identified for the first time from the total saponins of Panax quinquefolium. The study shows that the HPD of the chiral structural modification has reached the level of industrialization, the pharmacological activity of anti cervical cancer is stronger and the toxicity is low, and the HPD injection made of it has a good development. The pharmacokinetics of a class of innovative drugs is one of the important contents of the development of innovative drugs. At present, the pharmacokinetic study of rats has not yet been reported. In this paper, the UPLC-MS/MS quantitative analysis method of HPD in rat plasma, tissue and excretion was established for the first time, and the HPD injection was given to rats by intravenous administration. The absorption, distribution and excretion of the rat in vivo, the plasma protein binding rate of HPD in vitro was measured. The UPLC-Q-TOF/MS qualitative analysis method of HPD metabolites in rats was established for the first time, and its metabolites were identified, and the metabolic rules of HPD in rats were revealed. The new results were obtained: 1, HPD injection's blood concentration time curve. The UPLC-MS/MS quantitative analysis of HPD in rat plasma was established for the first time. The concentration of drug in plasma at different time points was measured after intravenous injection of HPD injection in rats, middle and high dose group (8.0,16.0,32.0mg/kg). The experimental data were calculated by DAS3.0 software, and the related parameters of the pharmacokinetics of HPD injection were obtained. Low dose group, T1/2 5.76 + 1.46h, Cmax5535.33 + 610.76 g/L, Tmax 0.10 + 0.00h, AUC (0-t) 11670.15 + 1364.30 micron h/L, AUC (0-) 12023.88 + 1573.57, 0.67 + 0.08. (0- infinity) 26986.39 + 4635.18 mu h/L, Vz 5.63 + 0.90L/kg, CLz 0.61 + 0.10L/h/kg, high dose group, T1/2 6.41 + 1.40h, Cmax 24683.57 + 3693.03 mu g/L, Tmax 0.10 + 5309.65 mu, 49165.27 + 5335.80 Mu Vz 5.52-6.07L/kg suggests not only in the blood, but also in the tissue; T1/2 5.76-6.50h and CLz 0.61-0.67L/h/kg suggest the slow.2 of HPD injection and the distribution of HPD injection for the first time to establish a UPLC-MS/MS quantitative analysis method for the determination of HPD in rat tissues. The content of HPD in 13 tissues (heart, lung, spleen, spleen, liver, kidney, kidney, intestines, intestines, intestines, muscles, fat, brain, uterus and testis) of 13 tissues (heart, lung, spleen, liver, kidney, intestine, intestines, muscle, fat, brain, uterus and testis) of 13 tissues of rats (15min, 1H, 4h, 6h) after intravenous administration of HPD injection in rats were measured. The results showed that HPD was widely distributed in various tissues, mainly in the kidney, uterus, and other groups. The distribution in the fabric was different with time; and the accumulation of.3 was not produced in the tissues. The plasma protein binding rate of HPD injection was studied for the first time by the UPLC-MS/MS quantitative analysis method of HPD in rat plasma, dog plasma, human plasma and dialysate. HPD and the above three species were determined by the equilibrium dialysis method. The protein binding rate of plasma was as follows: the protein binding rates of 0.1 mu g/m L, 4 mu g/m L and 16 mu g/m L were 68.36%, 69.07% and 70.48%, respectively, and protein binding rates with dog plasma were 67.66%, 74.42% and 72.32%, respectively. The protein binding rate with human plasma was 65.19%, 68.40% and 66.09%. results, respectively, in 0.1 mu. In the range of g/m L-16.0 g/m L, the protein binding rate of HPD and the above three plasma was not dependent on concentration; the protein binding rate of the three plasma was not significantly different from that of.4. The UPLC-MS/MS determination method of HPD in rat bile, urine and feces was established for the first time in the excretory study of HPD injection. The intravenous administration of 16 in rats was respectively given to 16. After HPD injection of mg/kg, the content of HPD in different time segments of bile, urine and feces of rats was found as follows: the cumulative excretion percentage of bile in 36h was only 0.06%, while the cumulative excretion percentage of urine and feces in 72h was 46.39% and 0.18%., respectively, indicating that the excretion of the prototype drug of HPD injection was very few in the bile and urine, mainly excreted by feces. The total cumulative excretion percentage of the three excreta was only 46.63%, indicating that the biotransformation in the rat was mainly metabolic elimination.5. The metabolic study of HPD injection was the first to establish the UPLC-Q-TOF/MS qualitative analysis method of HPD metabolites in rat bile, urine and feces. The HPD injection of 16.0mg/kg in rats was identified by intravenous injection. After the liquid, the metabolites of rat dung, urine and bile in 72h were obtained. The results were as follows: 21 metabolites were detected in HPD injection in rats except the original medicine, of which 13 metabolites were detected in the feces and urine, 12 were phase I metabolites, and 8 metabolites were detected in the bile samples, and 5 were metabolites of the second phase. The results showed that HPD metabolism and phase II metabolism occurred simultaneously in rats, and the phase I metabolites were mainly in the feces and urine, and the phase II was mainly in the bile.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R969.1

【参考文献】