CP-25在大鼠体内药代动力学及组织分布的研究
发布时间：2019-06-21 19:33
【摘要】：白芍总苷(Total glucoside of paeony,TGP)是白芍根部中提取的有效成分。TGP作为抗炎免疫调节药于1998年被中国食品药品监督管理局批准上市,用于治疗类风湿性关节炎(rheumatoid arthritis,RA)有较好的治疗效果,在长期的临床使用中没有明显的不良反应。但是,起效慢和生物利用度低等原因限制了TGP在临床的应用。芍药苷(Paeoniflorin,Pae),是一种水溶性的单帖苷类,是TGP的主要有效成分,含量超过总苷的40%。课题组已研究证明Pae有较好的抗炎效果和调节免疫。然而,Pae生物利用度(3%-4%)较低,口服吸收较差。因此,课题组前期对Pae的6位羟基进行酯化修饰,得到新的化合物苯磺酰芍药苷(benzenesulfonyl paeoniflorin,代号:CP-25)。目前课题组研究证实了CP-25拥有相比于Pae更好的抗炎活性,CP-25的吸收机制主要是被动吸收,在大鼠小肠中CP-25的吸收要明显高于Pae,同时CP-25在大鼠体内的口服生物利用度要高于Pae(10.6%)。性别、食物和疾病等因素对药物的药代动力学性质有明显的影响。因此,本研究主要考察性别、食物和佐剂型关节炎(adjuvant arthritis,AA)对CP-25在大鼠体内药代动力学影响及CP-25组织分布的研究。目的:1.考察性别、食物和AA等因素对CP-25及其血浆代谢物Pae(M1)在大鼠体内药代动力学的影响。2.考察CP-25和Pae在大鼠体内组织分布的差异,对比CP-25在雌雄大鼠的组织分布中的差异。方法:1.建立了CP-25和Pae在生物样品(血液和组织)中检测的超高效液相色谱串联质谱(ultra-performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)方法。2.单次灌胃CP-25(50 mg·kg-1),对比大鼠在不同的状态下(雌/雄,喂食/禁食,健康/疾病,单次/连续多次给药),CP-25和M1药代动力学的差异。3.组织分布研究中,在正常大鼠连续多次灌胃给药(50 mg·kg-1)CP-25和Pae后,取其心、肝、脾、肺、肾、小肠、胃、脂肪、肌肉、脑、颌下腺和滑膜组织,测其药物浓度。结果:1.CP-25、Pae和内标(internal standard,IS)的出峰时间分别为1.84,0.46和1.49min,出峰时间适宜,无明显的大鼠血浆内源性物质的干扰。CP-25和Pae的标准曲线分别为y=0.0056x-0.0122(r2=0.9990)和y=0.0049x-0.0148(r2=0.9990),浓度范围为2~800 ng·m L-1,线性关系良好;CP-25和Pae定量下限为2 ng·m L-1,其浓度适用于后续的药物检测;CP-25和Pae的日内和日间精密度和准确度均在15%以内;CP-25和Pae的回收率的范围分别为88.5%~102.9%和99.5%~112.4%,IS(200 ng·m L-1)的回收率为103.0±3.3%。CP-25和Pae的基质效应的范围分别为100.9%~113.7%和102.3%~113.8%,IS(200 ng·m L-1)的基质效应为99.7±4.4%;CP-25和Pae在不同条件下均较稳定。2.性别对CP-25药代动力学的影响:CP-25单次灌胃给药后(50 mg·kg-1),其表观分布容积(V)在雌雄大鼠中有显著性差异(P0.05)。达峰时间(Tmax)在2-3h之间,分布半衰期(t1/2α)和消除半衰期(t1/2β)分别是1-2h和7-8h。M1药代动力学参数在雌雄大鼠中没有显著性差异。3.食物对CP-25药代动力学的影响:单次灌胃给药CP-25(50 mg·kg-1),禁食组和喂食组的药代动力学参数对比如下:清除率(CL)(CP-25:19.51±2.32 L·h·-1kg-1versus 16.55±0.58 L·h·-1kg-1;M1:23.31±2.58 L·h·-1kg-1 versus 16.40±2.92L·h·-1kg-1),禁食组CL明显低于喂食组(P0.05)。药时曲线下面积(AUC(0-∞))(CP-25:2892.48±89.53μg·L-1·h versus 3024.34±107.15μg·L-1·h;M1:2166.64±247.61μg·L-1·h versus 3137.30±584.25μg·L-1·h),禁食组AUC(0-∞)明显低于喂食组(P0.05)。此外,M1的AUC(0-t)(2062.02±206.48μg·L-1·h versus2871.64±409.82μg·L-1·h)和达峰浓度(Cmax)(233.36±32.67μg·L-1versus322.00±45.22μg·L-1),喂食组明显高于禁食组(P0.05)。4.AA对CP-25药代动力学的影响:在AA状态下,CP-25的AUC明显低于在正常大鼠(AUC(0-t),2461.90±168.25μg·L-1*h versus 2727.59±215.01μg·L-1·h;AUC(0-∞),2685.91±60.97 ug·L-1·h versus 2892.48±89.53 ug·L-1·h)(P0.05)。M1在AA状态下的药代动力学参数V明显高于正常大鼠(127.18±21.24 L·kg-1 versus96.55±9.76 L·kg-1,P0.05)。5.多次给药对CP-25药代动力学的影响:AUC和达峰浓度(Cmax)相比于单次给药有显著性差异(AUC(0-t),2461.90±168.25μg·L-1·h versus 2856.62±263.54μg·L-1·h;AUC(0-∞),2685.91±60.97μg·L-1·h versus 2909.34±302.84μg·L-1·h;Cmax,354.14±23.62μg·L-1 versus 392.82±10.69μg·L-1)(P0.05)。此外,CP-25的稳态药时曲线下面积(AUCss),稳态血药浓度(Cav)和波动系数(undulate factors,DF)分别为2512.02±114.83μg·L-1·h,209.34±9.57μg·L-1和1.88±0.07。6.CP-25和Pae在大鼠体内的组织分布:CP-25在大鼠组织中有较广泛的分布,存在雌雄差异。雄性大鼠的肝、滑膜、肌肉、小肠和脾脏中有较高的药物浓度,靶组织滑膜中在3h发现较高的药物浓度。在雌性大鼠中,在肝脏、小肠、肌肉和脑中CP-25浓度较高,其它组织均有一定的分布。同时,对比Pae,CP-25在组织中的药物浓度均高于Pae,其中脑中的CP-25浓度要明显高于Pae。结论:1.CP-25和M1在雌雄大鼠体内表观分布容积有显著性差异,其它药代动力学参数没有显著的药代动力学差异,2.食物的摄取会增加CP-25在大鼠体内的吸收,减慢其清除率。3.AA会降低CP-25在大鼠体内的吸收。4.CP-25和Pae在大鼠体内组织分布存在着广泛的差异。在雌雄大鼠体内,CP-25主要是肝、肺、脑、滑膜和小肠的差异。
[Abstract]:The total amount of Radix Paeoniae Alba (TGP) is the effective component extracted from the root of Radix Paeoniae Alba. TGP, as an anti-inflammatory and immunomodulating drug, was approved by the China Food and Drug Administration in 1998 for the treatment of rheumatoid arthritis (RA), and has no obvious adverse reaction in the long-term clinical use. However, the causes of slow onset and low bioavailability limit the application of TGP in clinical applications. Paeonifloin, Pae, is a water-soluble monotype, which is the main active ingredient of TGP, with a content of more than 40% of the total content. The research group has studied that Pae has a good anti-inflammatory effect and the regulation of immunity. However, Pae bioavailability (3%-4%) is low and oral absorption is poor. In the early stage of the research group, the 6-hydroxy group of Pae was subjected to the esterification modification to obtain the new compound, benzenesulfonyl paeonifloin, code: CP-25. At present, the research group has confirmed that CP-25 has better anti-inflammatory activity than Pae, and the absorption mechanism of CP-25 is mainly passive absorption, and the absorption of CP-25 in the small intestine of rats is significantly higher than Pae, while the oral bioavailability of CP-25 in rats is higher than Pae (10.6%). Factors such as gender, food and disease have a clear effect on the pharmacokinetic properties of the drug. In this study, the effects of gender, food and adjuvant arthritis (AA) on the pharmacokinetics of CP-25 in rats and the distribution of CP-25 were studied. Objective:1. The effect of gender, food and AA on the pharmacokinetics of CP-25 and its plasma metabolite Pae (M1) in rats was investigated. The differences of the distribution of CP-25 and Pae in the tissues of the rats were examined, and the difference of CP-25 in the tissue distribution of male and female rats was compared. Method:1. An ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/ MS) method for the detection of CP-25 and Pae in biological samples (blood and tissue) was established. The difference in the pharmacokinetics of CP-25 and M1 was compared with a single Gavage of CP-25 (50 mg 路 kg-1) compared to the rats in different conditions (female/ male, feeding/ fasting, health/ disease, single/ continuous multiple-dose). In the study of tissue distribution, after the normal rats were given intragastric administration (50 mg 路 kg-1) CP-25 and Pae, the heart, the liver, the spleen, the lung, the kidney, the small intestine, the stomach, the fat, the muscle, the brain, the submandibular gland and the synovial tissue were taken to measure the drug concentration. Results:1. The peak-out time of CP-25, Pae and internal standard (IS) was 1.84, 0.46 and 1.49 min, respectively. The standard curves of CP-25 and Pae are y = 0.0056x-0.0122 (r2 = 0.9990) and y = 0.0049x-0.0148 (r2 = 0.9990), the concentration range is 2-800ng 路 mL-1, the linear relationship is good, the lower limit of CP-25 and Pae is 2 ng 路 m L-1, the concentration of CP-25 and Pae is within 15%; The recoveries of CP-25 and Pae were 88.5%-102.9% and 99.5%-112.4%, respectively. The recovery of IS (200 ng 路 m-L-1) was 103.0-3.3%. The matrix effects of CP-25 and Pae were 100.9%-113.7% and 102.3%-113.8%, respectively. The matrix effect of IS (200 ng 路 m-L-1) was 99.7-4.4%, and CP-25 and Pae were more stable under different conditions. The effect of gender on the pharmacokinetics of CP-25: The apparent distribution volume (V) of CP-25 after a single oral administration (50 mg 路 kg-1) was significantly different in male and female rats (P0.05). The peak-to-peak time (Tmax) was between 2 and 3 h, the distribution half-life (t1/2) and the elimination half-life (t1/2) were 1-2 h and 7-8 h, respectively. The pharmacokinetic parameters of M1 were not significantly different in male and female rats. The effect of food on the pharmacokinetics of CP-25: The pharmacokinetic parameters of CP-25 (50 mg 路 kg-1), fasting group and feeding group were as follows: clearance (CL) (CP-25: 19.51-2.32 L 路 h 路 -1 kg-1 reverse 16.55-0.58 L 路 h 路 -1 kg-1; M1: 23.31 mg 2.58 L 路 h 路 -1 kg-1 versus 16.40 (2.92 L 路 h 路 -1 kg -1), and the fasting group CL was significantly lower than that of the feeding group (P0.05). The area under the curve (AUC (0-1)) (CP-25: 2892.48, 89.53. mu.g/ L-1 路 h, M1: 2166.64-247.61. mu.g 路 L-1 路 h versus 3137.30-584.25. mu.g 路 L-1 路 h) and the AUC (0-1) of the fasting group were significantly lower than that of the feeding group (P0.05). In addition, the AUC (0-t) of M1 (2062.02-206.48. mu.g 路 L-1 路 h versus2871.64-409.82. mu.g 路 L-1 路 h) and the peak-to-peak concentration (Cmax) (233.36-32.67. mu.g 路 L-1 vers33.2.00-45.22. mu.g 路 L-1) were significantly higher in the feeding group than in the fasted group (P0.05). The AUC of CP-25 was significantly lower than in normal rats (AUC (0-t), 2461.90, and 25.mu. g 路 L-1 * h versus 2727.59-215.01. mu.g 路 L-1 路 h; AUC (0-1), 2685.91 g 路 L-1 路 h versus 2892.48 (89.53 ug 路 L-1 路 h) (P0.05). The pharmacokinetic parameters of M1 in AA state were significantly higher than that of normal rats (127.18-21.24 L 路 kg-1 versus6.55 and 9.76 L 路 kg-1, P0.05). The effect of multiple administrations on the pharmacokinetics of CP-25: the AUC and the peak concentration (Cmax) were significantly different from single-dose (AUC (0-t), 2461.90, and 25.mu. g 路 L-1 路 h versus 2856.62-263.54. mu.g 路 L-1 路 h; AUC (0-1), 2685.91-60.97. mu.g 路 L-1 路 h versus 2909.34-302.84. mu.g 路 L-1 路 h; Cmax, 354.14 to 23.62. m u.g 路 L-1 versus 392.82 (10.69. mu.g 路 L-1) (P0.05). In addition, the area under the steady-state curve of CP-25 (AUCss), steady-state plasma concentration (Cav) and fluctuation factors (DF) were 2512.02-114.83. m u.g 路 L-1 路 h, 209.34-9.57. mu.g 路 L-1 and 1.88-0.07.6.6. CP-25 and Pae were distributed in the rat. There were male and female differences. The liver, synovium, muscle, small intestine and spleen of male rats had higher drug concentration, and higher drug concentration was found in the synovium of the target tissue at 3 h. In female rats, the concentration of CP-25 in the liver, the small intestine, the muscle and the brain is high, and other tissues have a certain distribution. At the same time, the drug concentration in the tissues of Pae and CP-25 was higher than Pae, and the concentration of CP-25 in the brain was significantly higher than Pae. Conclusion:1. There is a significant difference in the apparent distribution volume of CP-25 and M1 in male and female rats, and there are no significant pharmacokinetic differences in other pharmacokinetic parameters. The uptake of the food increases the absorption of CP-25 in the rat, and decreases its clearance.3. AA decreases the absorption of CP-25 in the rat.4. There is a wide range of differences in the tissue distribution of CP-25 and Pae in the rat. In male and female rats, CP-25 is mainly the difference of liver, lung, brain, synovium and small intestine.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
本文编号：2504298
[Abstract]:The total amount of Radix Paeoniae Alba (TGP) is the effective component extracted from the root of Radix Paeoniae Alba. TGP, as an anti-inflammatory and immunomodulating drug, was approved by the China Food and Drug Administration in 1998 for the treatment of rheumatoid arthritis (RA), and has no obvious adverse reaction in the long-term clinical use. However, the causes of slow onset and low bioavailability limit the application of TGP in clinical applications. Paeonifloin, Pae, is a water-soluble monotype, which is the main active ingredient of TGP, with a content of more than 40% of the total content. The research group has studied that Pae has a good anti-inflammatory effect and the regulation of immunity. However, Pae bioavailability (3%-4%) is low and oral absorption is poor. In the early stage of the research group, the 6-hydroxy group of Pae was subjected to the esterification modification to obtain the new compound, benzenesulfonyl paeonifloin, code: CP-25. At present, the research group has confirmed that CP-25 has better anti-inflammatory activity than Pae, and the absorption mechanism of CP-25 is mainly passive absorption, and the absorption of CP-25 in the small intestine of rats is significantly higher than Pae, while the oral bioavailability of CP-25 in rats is higher than Pae (10.6%). Factors such as gender, food and disease have a clear effect on the pharmacokinetic properties of the drug. In this study, the effects of gender, food and adjuvant arthritis (AA) on the pharmacokinetics of CP-25 in rats and the distribution of CP-25 were studied. Objective:1. The effect of gender, food and AA on the pharmacokinetics of CP-25 and its plasma metabolite Pae (M1) in rats was investigated. The differences of the distribution of CP-25 and Pae in the tissues of the rats were examined, and the difference of CP-25 in the tissue distribution of male and female rats was compared. Method:1. An ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/ MS) method for the detection of CP-25 and Pae in biological samples (blood and tissue) was established. The difference in the pharmacokinetics of CP-25 and M1 was compared with a single Gavage of CP-25 (50 mg 路 kg-1) compared to the rats in different conditions (female/ male, feeding/ fasting, health/ disease, single/ continuous multiple-dose). In the study of tissue distribution, after the normal rats were given intragastric administration (50 mg 路 kg-1) CP-25 and Pae, the heart, the liver, the spleen, the lung, the kidney, the small intestine, the stomach, the fat, the muscle, the brain, the submandibular gland and the synovial tissue were taken to measure the drug concentration. Results:1. The peak-out time of CP-25, Pae and internal standard (IS) was 1.84, 0.46 and 1.49 min, respectively. The standard curves of CP-25 and Pae are y = 0.0056x-0.0122 (r2 = 0.9990) and y = 0.0049x-0.0148 (r2 = 0.9990), the concentration range is 2-800ng 路 mL-1, the linear relationship is good, the lower limit of CP-25 and Pae is 2 ng 路 m L-1, the concentration of CP-25 and Pae is within 15%; The recoveries of CP-25 and Pae were 88.5%-102.9% and 99.5%-112.4%, respectively. The recovery of IS (200 ng 路 m-L-1) was 103.0-3.3%. The matrix effects of CP-25 and Pae were 100.9%-113.7% and 102.3%-113.8%, respectively. The matrix effect of IS (200 ng 路 m-L-1) was 99.7-4.4%, and CP-25 and Pae were more stable under different conditions. The effect of gender on the pharmacokinetics of CP-25: The apparent distribution volume (V) of CP-25 after a single oral administration (50 mg 路 kg-1) was significantly different in male and female rats (P0.05). The peak-to-peak time (Tmax) was between 2 and 3 h, the distribution half-life (t1/2) and the elimination half-life (t1/2) were 1-2 h and 7-8 h, respectively. The pharmacokinetic parameters of M1 were not significantly different in male and female rats. The effect of food on the pharmacokinetics of CP-25: The pharmacokinetic parameters of CP-25 (50 mg 路 kg-1), fasting group and feeding group were as follows: clearance (CL) (CP-25: 19.51-2.32 L 路 h 路 -1 kg-1 reverse 16.55-0.58 L 路 h 路 -1 kg-1; M1: 23.31 mg 2.58 L 路 h 路 -1 kg-1 versus 16.40 (2.92 L 路 h 路 -1 kg -1), and the fasting group CL was significantly lower than that of the feeding group (P0.05). The area under the curve (AUC (0-1)) (CP-25: 2892.48, 89.53. mu.g/ L-1 路 h, M1: 2166.64-247.61. mu.g 路 L-1 路 h versus 3137.30-584.25. mu.g 路 L-1 路 h) and the AUC (0-1) of the fasting group were significantly lower than that of the feeding group (P0.05). In addition, the AUC (0-t) of M1 (2062.02-206.48. mu.g 路 L-1 路 h versus2871.64-409.82. mu.g 路 L-1 路 h) and the peak-to-peak concentration (Cmax) (233.36-32.67. mu.g 路 L-1 vers33.2.00-45.22. mu.g 路 L-1) were significantly higher in the feeding group than in the fasted group (P0.05). The AUC of CP-25 was significantly lower than in normal rats (AUC (0-t), 2461.90, and 25.mu. g 路 L-1 * h versus 2727.59-215.01. mu.g 路 L-1 路 h; AUC (0-1), 2685.91 g 路 L-1 路 h versus 2892.48 (89.53 ug 路 L-1 路 h) (P0.05). The pharmacokinetic parameters of M1 in AA state were significantly higher than that of normal rats (127.18-21.24 L 路 kg-1 versus6.55 and 9.76 L 路 kg-1, P0.05). The effect of multiple administrations on the pharmacokinetics of CP-25: the AUC and the peak concentration (Cmax) were significantly different from single-dose (AUC (0-t), 2461.90, and 25.mu. g 路 L-1 路 h versus 2856.62-263.54. mu.g 路 L-1 路 h; AUC (0-1), 2685.91-60.97. mu.g 路 L-1 路 h versus 2909.34-302.84. mu.g 路 L-1 路 h; Cmax, 354.14 to 23.62. m u.g 路 L-1 versus 392.82 (10.69. mu.g 路 L-1) (P0.05). In addition, the area under the steady-state curve of CP-25 (AUCss), steady-state plasma concentration (Cav) and fluctuation factors (DF) were 2512.02-114.83. m u.g 路 L-1 路 h, 209.34-9.57. mu.g 路 L-1 and 1.88-0.07.6.6. CP-25 and Pae were distributed in the rat. There were male and female differences. The liver, synovium, muscle, small intestine and spleen of male rats had higher drug concentration, and higher drug concentration was found in the synovium of the target tissue at 3 h. In female rats, the concentration of CP-25 in the liver, the small intestine, the muscle and the brain is high, and other tissues have a certain distribution. At the same time, the drug concentration in the tissues of Pae and CP-25 was higher than Pae, and the concentration of CP-25 in the brain was significantly higher than Pae. Conclusion:1. There is a significant difference in the apparent distribution volume of CP-25 and M1 in male and female rats, and there are no significant pharmacokinetic differences in other pharmacokinetic parameters. The uptake of the food increases the absorption of CP-25 in the rat, and decreases its clearance.3. AA decreases the absorption of CP-25 in the rat.4. There is a wide range of differences in the tissue distribution of CP-25 and Pae in the rat. In male and female rats, CP-25 is mainly the difference of liver, lung, brain, synovium and small intestine.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
【参考文献】
相关期刊论文 前6条
1 QIU Shi;SUN Hui;ZHANG Ai-Hua;XU Hong-Ying;YAN Guang-Li;HAN Ying;WANG Xi-Jun;;Natural alkaloids: basic aspects, biological roles, and future perspectives[J];Chinese Journal of Natural Medicines;2014年06期
2 韩丽;张玲非;张肖玲;王鑫国;牛丽颖;;白芍总苷在免疫性肝损伤小鼠药代动力学研究[J];中药药理与临床;2012年06期
3 李培培;解国雄;宋珊珊;黄蓓;吴育晶;汪庆童;常艳;张运芳;周爱武;刘丽华;张玲玲;魏伟;;大鼠佐剂性关节炎模型表现特征及评价指标[J];中国免疫学杂志;2012年05期
4 钟大放;李高;刘昌孝;;生物样品定量分析方法指导原则(草案)[J];药物评价研究;2011年06期
5 张洪峰;肖卫国;侯平;;白芍总苷治疗系统性红斑狼疮的临床研究[J];中国中西医结合杂志;2011年04期
6 张洪峰;侯平;肖卫国;;白芍总苷治疗非系统受累干燥综合征的临床观察[J];中国中西医结合杂志;2007年07期
，本文编号：2504298
本文链接：https://www.wllwen.com/shoufeilunwen/mpalunwen/2504298.html
最近更新
教材专著
