溶栓候选药物FGFC1药物代谢动力学特性的研究

发布时间：2018-01-08 19:08

本文关键词：溶栓候选药物FGFC1药物代谢动力学特性的研究　出处：《上海海洋大学》2016年硕士论文　论文类型：学位论文

【摘要】：随着气候变暖、资源匮乏和人口增多等变化,心脑血管疾病(cardiovascular and cerebrovascular diseases,CCVD)、癌症、病毒性感染等难治性疾病严重威胁到人类的健康和生存,尤其是心脑血管疾病药物已经成为世界各国药物科学家研究的热点,发现新型海洋药物有可能彻底治疗难治性疾病逐渐成为社会共识。论文基于海洋真菌长孢葡萄穗霉菌FG216(Stachybotrys longispora FG216)代谢产物溶栓候选药物FGFC1的结构和纤溶活性的确认,研究了比格犬体内的药物代谢动力学特性和组织分布规律,探索了FGFC1在CACO-2细胞模型中的吸收转运特性及其对肝药酶活性的影响,在小分子溶栓海洋新药的开发上具有应用价值,在新型溶栓理论的建立上具有科学意义。第一章综述了国内外溶栓药物的研究现状,同时介绍了以同位素标记法和代谢组学为代表的体内药物代谢动力学以及CACO-2细胞模型和肝微粒体酶为代表的体外药物代谢动力学。第二章是溶栓候选药物FGFC1的结构和纤溶活性的确认。溶栓候选药物FGFC1结构通过1H NMR,13C NMR等光谱被确定为2,5-二((2-(2,4,-二甲基-2,4-二烯)壬基)-(2-甲基-3-羟基)-吡喃并(5-羟基)-异吲哚酮基)-戊酸。通过Glu-纤维蛋白溶酶原和Lys-纤维蛋白溶酶原测量体外活性,结果表明0.1-0.4 mmol/L FGFC1能提高纤维蛋白溶酶活性2.05-11.44倍。FITC-纤维蛋白原降解实验表明FGFC1通过激活纤维蛋白溶酶原的活性来发挥作用。剂量为10 mg/kg的FGFC1能降解大鼠急性肺血栓。第三章是研究FGFC1在比格犬体内的药物代谢动力学和组织分布。比格犬静脉注射7.5 mg·kg-1、5.0 mg·kg-1和2.5mg·kg-1三种剂量的FGFC1,在不同的时间点取血,用HPLC检测血浆和组织器官中FGFC1的浓度并通过PKslover软件计算其药物代谢动力学参数,统计组织分布规律。药物代谢动力学实验结果表明,FGFC1在比格犬体内消除半衰期(t1/2β)分别为49.035±2.171,48.422±2.113和48.811±2.372 min;达峰浓度Cmax分别为56.48±6.23、48.63±5.53和13.64±2.76μg?mL-1;机体总消除率(CL)分别为0.0062±0.0004、0.0071±0.0008和0.0092±0.0006(L·min-1·kg-1);平均保留时间(MRT)分别为28.17±1.16、26.23±0.35和28.66±0.84min。心、肝、脾、肺、肾、脑、肌肉、睾丸、肠、胆汁、粪便和尿液中均发现FGFC1原型药,其中肝脏和胆汁中的含量最高,其中肝脏和胆汁中含量较高,肺、肾脏、脾脏、胃粪便、尿液中含量中等,其余含量一般。表明FGFC1在全身各处都能检测出FGFC1,说明FGFC1具有良好的组织分布,对达到全身各个病灶第四章是研究FGFC1吸收和转运特性。通过LC/MS/MS技术检测FGFC1在CACO-2细胞模型的外排比、总回收率和表观渗透系数。结果显示AP→BL方向和BL→AP方向的Papp值均小于2.5×10-6 cm·s-1,从AP→BL侧的Papp AP-BL值和BL→AP侧的Papp BL-AP值没有统计学意义上的差异,且其外排比值均接近1.5,说明FGFC1在CACO-2细胞模型中以被动扩散为主。口服给药FGFC1可能吸收不完全,使用静脉给药被认为能增强其生物利用度。第五章是研究FGFC1对大鼠肝微粒体酶的影响。在大鼠肝微粒体孵育体系加入探针底物非那西丁、咪达唑仑及氯唑沙宗来反映FGFC1对大鼠肝微粒体酶细胞色素CYP450(Cytochrome P450 proteins,CYP450)的影响,同时,还观察了在连续给药下大鼠的肝脏组织形态学和大鼠与大白兔的心电图参数。实验结果显示,FGFC1在41.6 mg·kg-1·d-1的高剂量下能显著诱导CYP3A4,还能带来肝窦扩张和细胞肿大等形态学上的变化。大白鼠和大白兔的处理剂量为7.5 mg·kg-1、5.0mg·kg-1和2.5mg·kg-1的FGFC1对其P波、PR间期、QRS波群、ST-T间期以及心率等心电图参数没有显著影响。FGFC1在连续高剂量给药下可能是一种潜在的CYP3A4诱导剂,同时FGFC1的长期给药可能对肝脏产生一定的影响,这需要以后继续进行研究。溶栓候选药物FGFC1的结构经NMR和MS等被确认为吲哚类化合物的2,5-二((2-(2,4,-二甲基-2,4-二烯)壬基)-(2-甲基-3-羟基)-吡喃并(5-羟基)-异吲哚酮基)-戊酸,纤溶活性被体内外实验所证实。FGFC1比格犬体内的半衰期为48-49min,属于二室模型,广泛分布于个组织和器官,主要分布于肝脏和胆汁。FGFC1从外排比、总回收率和表观渗透系数等体现出以被动扩散为主,高剂量FGFC1能显著诱导肝药酶CYP3A4并且还能带来肝窦扩张和细胞肿大等形态学上的变化。但不会得心脏功能产生影响。
[Abstract]:With climate warming, resources shortage and the increasing population changes, cardiovascular and cerebrovascular diseases (cardiovascular and, cerebrovascular diseases, CCVD), cancer, viral infections and other refractory disease is a serious threat to human health and survival, especially of drugs for cardiovascular and cerebrovascular diseases has become the focus of pharmaceutical scientists around the world, find new marine drugs may complete the treatment of refractory disease has gradually become a social consensus. Based on the marine fungus spore fungus Stachybotrys FG216 long (Stachybotrys longispora FG216) fiber structure and metabolites of thrombolytic drug candidates FGFC1 dissolution activity confirmed of beagle dogs pharmacokinetic characteristics and tissue distribution, to explore the FGFC1 absorption and transportation characteristics in CACO-2 cell model and its effect on the activity of liver enzyme, in small molecule drug development on marine thrombolysis should have With the value of scientific significance in the establishment of new thrombolytic theory. The first chapter reviews the domestic and foreign research status of thrombolytic drugs, and introduces the in vitro pharmacokinetics by isotope labeling method and metabolomics pharmacokinetics as the representative and the CACO-2 cell model and liver microsomal enzyme as the representative of the second chapter is confirmed. The fiber structure and thrombolytic drug candidate FGFC1 fibrinolytic activity. Thrombolytic drug candidate FGFC1 13C NMR structure by 1H NMR spectra was determined to be two 2,5- ((2- (2,4, two - methyl nonyl diene -2,4-) - (2-) -3- hydroxy methyl) - pyrano (5- hydroxy) - isoindolone base) - pentanoic acid by Glu- plasminogen and Lys- plasminogen in vitro activity measurements, the results show that 0.1-0.4 mmol/L FGFC1 can increase the soluble enzyme activity 2.05-11.44 times.FITC- fibrinogen degradation experiment showed that FGFC1 induced by fibrin Live plasminogen activity to play a role. The dose of 10 mg/kg FGFC1 of acute pulmonary embolism in rats. The third chapter is the degradation of FGFC1 in beagle dogs in vivo pharmacokinetics and tissue distribution of beagle dogs. Intravenous injection of 7.5 mg - kg-1,5.0 Mg - kg-1 and 2.5mg - kg-1 three dose of FGFC1. The blood samples were taken at different time points, with the concentration of FGFC1 in plasma and tissue organ detection in HPLC and through the PKslover software to calculate the pharmacokinetic parameters, statistical tissue distribution. The experimental results show that the pharmacokinetics of FGFC1 in beagle dogs, the elimination half-life (t1/2 beta) were 49.035 + 2.171,48.422 + 2.113 and 48.811 + 2.372 min; peak concentration Cmax = 56.48 + 6.23,48.63 + 5.53 and 13.64 + 2.76 G? ML-1; the total elimination rate (CL) were 0.0062 + 0.0004,0.0071 + 0.0008 and 0.0092 + 0.0006 (L min-1 kg-1); the average. Retention time (MRT) were 28.17 + 1.16,26.23 + 0.35 and 28.66 + 0.84min. in heart, liver, spleen, lung, kidney, brain, testis, muscle, intestine, bile, FGFC1 drug prototype were found in the feces and urine, the content of liver and bile in the highest, higher content of liver and bile in the lung the spleen, stomach, kidney, feces, urine content of medium, the general content. The results indicated that FGFC1 can detect FGFC1 in the body, indicating that FGFC1 has good tissue distribution, to achieve the body of each lesion fourth chapter is the research on FGFC1 uptake and transport characteristics. Through the detection of FGFC1 LC/MS/MS in CACO-2 cell model of parallelism the total recovery rate, and the apparent permeability coefficient. The results showed that AP, BL, AP direction and BL direction of the Papp values were less than 2.5 * 10-6 cm s-1, from AP to BL Papp AP-BL and BL side, AP side of the Papp BL-AP value was no significant difference in meaning, and the external value of parallelism Close to 1.5, indicating that FGFC1 is dominated by passive diffusion in CACO-2 cell model. The oral administration of FGFC1 could not completely absorbed, the use of intravenous administration is thought to enhance its bioavailability. The fifth chapter is the research on the influence of FGFC1 on rat liver microsomal enzymes. Education system added to probe substrate phenacetin in rat liver microsomes then, midazolam and chlorazol Sha Zong to reflect on the pigment FGFC1 in rat liver microsome cell CYP450 (Cytochrome P450 proteins, CYP450) effect, at the same time, was also observed in the continuous ECG parameters of liver tissue morphology medicine rats and rats and white rabbits. Experimental results show that high dose of FGFC1 in 41.6 Mg - kg-1 - D-1 could significantly induce CYP3A4, but also bring the changes of hepatic sinus expansion and enlargement of cell morphology. Treatment dose of rats and rabbits was 7.5 mg - kg-1,5.0mg - kg-1 and 2.5mg - kg-1 to FGFC1 The P wave, QRS wave, PR interval, ST-T interval and heart rate did not significantly affect the.FGFC1 ECG parameters in continuous high dose may be a potential inducer of CYP3A4, FGFC1 and the long-term administration may have a certain impact on the liver, the need to continue after thrombolysis in research. The structure of candidate drug FGFC1 by NMR and MS were identified as indole compounds (2,5- two (2- (2,4, two - -2,4- diene nonyl methyl) - (2-) -3- hydroxy methyl (5-) - pyran and hydroxyl) - isoindole ketone) - pentanoic acid, fibrinolytic activity was confirmed by in vitro and in vivo..FGFC1 Beagle half-life is 48-49min, which belongs to the two compartment model, widely distributed in tissues and organs, mainly distributed in liver and bile.FGFC1 from the outer parallelism, the total recovery rate and the apparent permeability coefficient shows by passive diffusion. High dose FGFC1 could significantly induce liver enzymes and CYP3A4 It also brings morphological changes, such as the dilatation of the hepatic sinusoids and the enlargement of the cells, but does not have the effect of the heart function.

【学位授予单位】：上海海洋大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R96

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