CP-25在大鼠體內(nèi)藥代動力學(xué)及組織分布的研究
發(fā)布時間：2019-06-21 19:33
【摘要】：白芍總苷(Total glucoside of paeony,TGP)是白芍根部中提取的有效成分。TGP作為抗炎免疫調(diào)節(jié)藥于1998年被中國食品藥品監(jiān)督管理局批準(zhǔn)上市,用于治療類風(fēng)濕性關(guān)節(jié)炎(rheumatoid arthritis,RA)有較好的治療效果,在長期的臨床使用中沒有明顯的不良反應(yīng)。但是,起效慢和生物利用度低等原因限制了TGP在臨床的應(yīng)用。芍藥苷(Paeoniflorin,Pae),是一種水溶性的單帖苷類,是TGP的主要有效成分,含量超過總苷的40%。課題組已研究證明Pae有較好的抗炎效果和調(diào)節(jié)免疫。然而,Pae生物利用度(3%-4%)較低,口服吸收較差。因此,課題組前期對Pae的6位羥基進(jìn)行酯化修飾,得到新的化合物苯磺酰芍藥苷(benzenesulfonyl paeoniflorin,代號:CP-25)。目前課題組研究證實(shí)了CP-25擁有相比于Pae更好的抗炎活性,CP-25的吸收機(jī)制主要是被動吸收,在大鼠小腸中CP-25的吸收要明顯高于Pae,同時CP-25在大鼠體內(nèi)的口服生物利用度要高于Pae(10.6%)。性別、食物和疾病等因素對藥物的藥代動力學(xué)性質(zhì)有明顯的影響。因此,本研究主要考察性別、食物和佐劑型關(guān)節(jié)炎(adjuvant arthritis,AA)對CP-25在大鼠體內(nèi)藥代動力學(xué)影響及CP-25組織分布的研究。目的:1.考察性別、食物和AA等因素對CP-25及其血漿代謝物Pae(M1)在大鼠體內(nèi)藥代動力學(xué)的影響。2.考察CP-25和Pae在大鼠體內(nèi)組織分布的差異,對比CP-25在雌雄大鼠的組織分布中的差異。方法:1.建立了CP-25和Pae在生物樣品(血液和組織)中檢測的超高效液相色譜串聯(lián)質(zhì)譜(ultra-performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)方法。2.單次灌胃CP-25(50 mg·kg-1),對比大鼠在不同的狀態(tài)下(雌/雄,喂食/禁食,健康/疾病,單次/連續(xù)多次給藥),CP-25和M1藥代動力學(xué)的差異。3.組織分布研究中,在正常大鼠連續(xù)多次灌胃給藥(50 mg·kg-1)CP-25和Pae后,取其心、肝、脾、肺、腎、小腸、胃、脂肪、肌肉、腦、頜下腺和滑膜組織,測其藥物濃度。結(jié)果:1.CP-25、Pae和內(nèi)標(biāo)(internal standard,IS)的出峰時間分別為1.84,0.46和1.49min,出峰時間適宜,無明顯的大鼠血漿內(nèi)源性物質(zhì)的干擾。CP-25和Pae的標(biāo)準(zhǔn)曲線分別為y=0.0056x-0.0122(r2=0.9990)和y=0.0049x-0.0148(r2=0.9990),濃度范圍為2~800 ng·m L-1,線性關(guān)系良好;CP-25和Pae定量下限為2 ng·m L-1,其濃度適用于后續(xù)的藥物檢測;CP-25和Pae的日內(nèi)和日間精密度和準(zhǔn)確度均在15%以內(nèi);CP-25和Pae的回收率的范圍分別為88.5%~102.9%和99.5%~112.4%,IS(200 ng·m L-1)的回收率為103.0±3.3%。CP-25和Pae的基質(zhì)效應(yīng)的范圍分別為100.9%~113.7%和102.3%~113.8%,IS(200 ng·m L-1)的基質(zhì)效應(yīng)為99.7±4.4%;CP-25和Pae在不同條件下均較穩(wěn)定。2.性別對CP-25藥代動力學(xué)的影響:CP-25單次灌胃給藥后(50 mg·kg-1),其表觀分布容積(V)在雌雄大鼠中有顯著性差異(P0.05)。達(dá)峰時間(Tmax)在2-3h之間,分布半衰期(t1/2α)和消除半衰期(t1/2β)分別是1-2h和7-8h。M1藥代動力學(xué)參數(shù)在雌雄大鼠中沒有顯著性差異。3.食物對CP-25藥代動力學(xué)的影響:單次灌胃給藥CP-25(50 mg·kg-1),禁食組和喂食組的藥代動力學(xué)參數(shù)對比如下: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),禁食組CL明顯低于喂食組(P0.05)。藥時曲線下面積(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),禁食組AUC(0-∞)明顯低于喂食組(P0.05)。此外,M1的AUC(0-t)(2062.02±206.48μg·L-1·h versus2871.64±409.82μg·L-1·h)和達(dá)峰濃度(Cmax)(233.36±32.67μg·L-1versus322.00±45.22μg·L-1),喂食組明顯高于禁食組(P0.05)。4.AA對CP-25藥代動力學(xué)的影響:在AA狀態(tài)下,CP-25的AUC明顯低于在正常大鼠(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狀態(tài)下的藥代動力學(xué)參數(shù)V明顯高于正常大鼠(127.18±21.24 L·kg-1 versus96.55±9.76 L·kg-1,P0.05)。5.多次給藥對CP-25藥代動力學(xué)的影響:AUC和達(dá)峰濃度(Cmax)相比于單次給藥有顯著性差異(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的穩(wěn)態(tài)藥時曲線下面積(AUCss),穩(wěn)態(tài)血藥濃度(Cav)和波動系數(shù)(undulate factors,DF)分別為2512.02±114.83μg·L-1·h,209.34±9.57μg·L-1和1.88±0.07。6.CP-25和Pae在大鼠體內(nèi)的組織分布:CP-25在大鼠組織中有較廣泛的分布,存在雌雄差異。雄性大鼠的肝、滑膜、肌肉、小腸和脾臟中有較高的藥物濃度,靶組織滑膜中在3h發(fā)現(xiàn)較高的藥物濃度。在雌性大鼠中,在肝臟、小腸、肌肉和腦中CP-25濃度較高,其它組織均有一定的分布。同時,對比Pae,CP-25在組織中的藥物濃度均高于Pae,其中腦中的CP-25濃度要明顯高于Pae。結(jié)論:1.CP-25和M1在雌雄大鼠體內(nèi)表觀分布容積有顯著性差異,其它藥代動力學(xué)參數(shù)沒有顯著的藥代動力學(xué)差異,2.食物的攝取會增加CP-25在大鼠體內(nèi)的吸收,減慢其清除率。3.AA會降低CP-25在大鼠體內(nèi)的吸收。4.CP-25和Pae在大鼠體內(nèi)組織分布存在著廣泛的差異。在雌雄大鼠體內(nèi),CP-25主要是肝、肺、腦、滑膜和小腸的差異。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5
本文編號：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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5
【參考文獻(xiàn)】
相關(guān)期刊論文 前6條
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 韓麗;張玲非;張肖玲;王鑫國;牛麗穎;;白芍總苷在免疫性肝損傷小鼠藥代動力學(xué)研究[J];中藥藥理與臨床;2012年06期
3 李培培;解國雄;宋珊珊;黃蓓;吳育晶;汪慶童;常艷;張運(yùn)芳;周愛武;劉麗華;張玲玲;魏偉;;大鼠佐劑性關(guān)節(jié)炎模型表現(xiàn)特征及評價指標(biāo)[J];中國免疫學(xué)雜志;2012年05期
4 鐘大放;李高;劉昌孝;;生物樣品定量分析方法指導(dǎo)原則(草案)[J];藥物評價研究;2011年06期
5 張洪峰;肖衛(wèi)國;侯平;;白芍總苷治療系統(tǒng)性紅斑狼瘡的臨床研究[J];中國中西醫(yī)結(jié)合雜志;2011年04期
6 張洪峰;侯平;肖衛(wèi)國;;白芍總苷治療非系統(tǒng)受累干燥綜合征的臨床觀察[J];中國中西醫(yī)結(jié)合雜志;2007年07期
，本文編號：2504298
本文鏈接：http://sikaile.net/shoufeilunwen/mpalunwen/2504298.html
最近更新
教材專著
