本文選題：HPD注射液 + UPLC-MS/MS�。� 參考：《吉林大學(xué)》2016年博士論文

【摘要】：羥基人參二醇(Hydroxylation panoxadiol,簡(jiǎn)稱HPD)是首次從西洋參莖葉總皂苷分離鑒定的一種三萜類新化合物。研究表明,手性結(jié)構(gòu)修飾方法制備的HPD達(dá)到了工業(yè)化水平;抗宮頸癌的藥理活性較強(qiáng);毒性較低;將其制成的HPD注射液具有良好開發(fā)一類創(chuàng)新藥物的前景,其中藥代動(dòng)力學(xué)是開發(fā)創(chuàng)新藥物的重要內(nèi)容之一,目前大鼠體內(nèi)的藥代動(dòng)力學(xué)研究尚未見報(bào)道。本論文首次建立了大鼠血漿、組織及排泄物中HPD的UPLC-MS/MS定量分析方法,研究了經(jīng)靜脈給予大鼠HPD注射液,藥物在大鼠體內(nèi)的吸收、分布和排泄規(guī)律,同時(shí)測(cè)定了HPD在體外的血漿蛋白結(jié)合率;首次建立了大鼠體內(nèi)HPD代謝物的UPLC-Q-TOF/MS定性分析方法,鑒定了其代謝產(chǎn)物,揭示了HPD在大鼠體內(nèi)的代謝規(guī)律。取得以下創(chuàng)新成果:1、HPD注射液的血藥濃度-時(shí)間曲線研究首次建立了大鼠血漿中HPD的UPLC-MS/MS定量分析方法。測(cè)定了經(jīng)靜脈給予大鼠低、中、高三個(gè)劑量組(8.0,16.0,32.0mg/kg)的HPD注射液后,不同時(shí)間點(diǎn)血漿中的藥物濃度。實(shí)驗(yàn)數(shù)據(jù)經(jīng)DAS3.0軟件計(jì)算,獲得的HPD注射液藥代動(dòng)力學(xué)的相關(guān)參數(shù)如下:低劑量組,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;中劑量組,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;高劑量組,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。結(jié)果表明,AUC、Cmax與HPD的給藥劑量均成正比例關(guān)系,符合一級(jí)動(dòng)力學(xué)特征;Vz 5.52-6.07L/kg提示不僅分布在血液中,在組織中也有分布;T1/2 5.76-6.50h及CLz 0.61-0.67L/h/kg提示HPD注射液的消除較慢。2、HPD注射液的分布研究首次建立了測(cè)定大鼠組織中HPD的UPLC-MS/MS定量分析方法。分別測(cè)定了大鼠靜脈給予16.0mg/kg的HPD注射液后,不同時(shí)間點(diǎn)(15min,1h,4h,6h)大鼠13個(gè)組織(心、肺、脾、肝、腎、小腸、大腸、胃、肌肉、脂肪、腦、子宮和睪丸)中HPD的含量。結(jié)果顯示,HPD快速?gòu)V泛地分布到各個(gè)組織中;主要分布在腎、子宮組織中,其它各個(gè)組織中的分布情況隨時(shí)間不同而各有差異;并且在各組織中均未產(chǎn)生蓄積作用。3、HPD注射液的血漿蛋白結(jié)合率研究首次建立了HPD在大鼠血漿、犬血漿、人血漿及透析液中的UPLC-MS/MS定量分析方法。結(jié)合平衡透析法,分別測(cè)定了HPD與上述三種血漿的蛋白結(jié)合率,結(jié)果如下:0.1μg/m L、4.0μg/m L和16μg/m L三個(gè)濃度的HPD與大鼠血漿的蛋白結(jié)合率分別為68.36%、69.07%和70.48%;與犬血漿的蛋白結(jié)合率分別為67.66%、74.42%和72.32%;與人血漿的蛋白結(jié)合率分別為65.19%、68.40%和66.09%。結(jié)果表明,在0.1μg/m L-16.0μg/m L的范圍內(nèi),HPD與上述三種血漿的蛋白結(jié)合率都不具有濃度依賴性;三種血漿的蛋白結(jié)合率種屬間無(wú)明顯差異。4、HPD注射液的排泄研究首次建立了大鼠膽汁、尿和糞樣中HPD的UPLC-MS/MS定量分析方法。分別測(cè)定了大鼠靜脈給予16.0mg/kg的HPD注射液后,大鼠膽汁、尿和糞樣中不同時(shí)間段HPD的含量,結(jié)果如下:膽汁在36h內(nèi)累積排泄百分比僅為0.06%,而尿和糞在72h內(nèi)累積排泄百分比分別為46.39%和0.18%。結(jié)果表明,HPD注射液的原型藥經(jīng)膽汁和尿的排泄極少,主要經(jīng)糞便排泄;由于三種排泄物中總的累積排泄百分比僅為46.63%,表明其在大鼠體內(nèi)的生物轉(zhuǎn)化是以代謝消除為主。5、HPD注射液的代謝研究首次建立了大鼠膽汁、尿和糞樣中HPD代謝物的UPLC-Q-TOF/MS定性分析方法。鑒定了大鼠經(jīng)靜脈給予16.0mg/kg的HPD注射液后,大鼠糞、尿和膽汁在72h內(nèi)的代謝產(chǎn)物,結(jié)果如下:HPD注射液在大鼠體內(nèi)除原形藥外,共檢測(cè)到21個(gè)代謝產(chǎn)物,其中糞樣和尿樣中共檢測(cè)出13個(gè)代謝產(chǎn)物,12個(gè)為Ⅰ相代謝產(chǎn)物;在膽汁樣品中共檢測(cè)出8個(gè)代謝產(chǎn)物,5個(gè)為Ⅱ相代謝產(chǎn)物。結(jié)果表明,HPD在大鼠體內(nèi)Ⅰ相代謝和Ⅱ相代謝會(huì)同時(shí)發(fā)生,而且Ⅰ相代謝產(chǎn)物主要發(fā)生在糞和尿中,Ⅱ相代物主要發(fā)生在膽汁中。
[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.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R969.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 許琪;李琳;左代英;張菁菁;李明曉;吳英良;;微透析技術(shù)結(jié)合HPLC法比較研究5-氟尿嘧啶在大鼠血液和腫瘤組織中的藥動(dòng)學(xué)[J];沈陽(yáng)藥科大學(xué)學(xué)報(bào);2015年07期

2 鄭鑫;李茜;尤海生;于佳;;培美曲賽二鈉與奧沙利鉑在Beagle犬體內(nèi)的藥動(dòng)學(xué)相互作用[J];西北藥學(xué)雜志;2015年02期

3 劉紅;張國(guó)楠;;宮頸癌化療耐藥相關(guān)問題[J];中國(guó)實(shí)用婦科與產(chǎn)科雜志;2015年03期

4 周暉;盧淮武;彭永排;林仲秋;;《2015年NCCN宮頸癌臨床實(shí)踐指南》解讀[J];中國(guó)實(shí)用婦科與產(chǎn)科雜志;2015年03期

5 荊凡波;徐文;龔偉玲;劉東華;隋忠國(guó);;靜脈注射烏苯美司/5-氟尿嘧啶在大鼠體內(nèi)的藥動(dòng)學(xué)研究[J];中國(guó)藥房;2015年01期

6 曹軍;何陽(yáng);劉洪強(qiáng);王賽博;趙保成;鄭曉輝;汪茂文;程英升;;吉西他濱及順鉑經(jīng)動(dòng)脈、靜脈注射后血漿、組織藥物濃度的變化[J];中國(guó)醫(yī)藥導(dǎo)報(bào);2014年32期

7 黃鑫;湯維維;江振洲;賀明;朱瑤俊;張陸勇;張尊建;;LC-MS/MS法測(cè)定Beagle犬血漿中紫杉醇及其藥代動(dòng)力學(xué)研究[J];藥物分析雜志;2013年11期

8 曾紅梅;陳萬(wàn)青;;中國(guó)癌癥流行病學(xué)與防治研究現(xiàn)狀[J];化學(xué)進(jìn)展;2013年09期

9 蔣玲燕;陳賽貞;田自有;徐利君;林佳苗;;格列齊特對(duì)氯沙坦鉀血漿蛋白結(jié)合率的影響[J];中國(guó)現(xiàn)代應(yīng)用藥學(xué);2013年09期

10 楊付英;張文萍;陳和莉;付巖;王欣瑜;魏世杰;楊小英;張宇馨;黨宏萬(wàn);;鹽酸伊立替康納米粒的藥動(dòng)學(xué)和組織分布[J];藥學(xué)學(xué)報(bào);2013年06期

相關(guān)博士學(xué)位論文 前2條

1 魯蓉;超聲彈性成像及綜合評(píng)分、液基薄層細(xì)胞學(xué)對(duì)宮頸病變?cè)\斷價(jià)值[D];中南大學(xué);2014年

2 劉鑫;5-氟尿嘧啶磁性脂質(zhì)體納米粒在肝癌動(dòng)物模型中的靶向性及藥動(dòng)學(xué)研究[D];中南大學(xué);2006年

相關(guān)碩士學(xué)位論文 前2條

1 孫帥;宮頸癌調(diào)強(qiáng)放療研究[D];中國(guó)協(xié)和醫(yī)科大學(xué);2007年

2 林能明;惡性腫瘤病人血漿中吉西他濱及其代謝產(chǎn)物的血藥濃度測(cè)定和藥代動(dòng)力學(xué)研究[D];浙江大學(xué);2004年

，

本文編號(hào)：1815918