Studies On The Pharmacokinetics Of XiaoShuan TongLuo Effective Components Group And The Related Compounds

Herbology is traditionally one of the more important modalities utilized in traditional Chinese Medicine,however,their unclear pharmacological mechanisms and the difficulties in quality control restrict their wider clinical use.Clarifying the effective compounds in Chinese multiherb remedy,which can be achieved by pharmacokinetic and metabolism study,has an important role in realizing the modernization,scientific research,and internationalization of traditional Chinese Medicine.XiaoShuanTongLuo,composed of eleven herbs(Milkvetch Root,Szechwan Lovage Rhizome,Danshen Root,Hawthorn Fruit,Sanchi,Turmeric Root Tuber,Common Aucklandia Root,Pagodatree Flower,Oriental Waterplantain Rhizome,Cassia Twig, Borneol) is a famous Chinese traditional prescription with better curative effect on cardiovascular diseases,acroparalysis and other symptoms arising from apoplexy.To understand the oral absorption and the metabolism of the active constituents of XiaoShuanTongLuo Effective Components Group,a specific and sensitive analytical method was established,to simultaneously determine rutin,hyperoside,quercetin and kaempferol,which were applied to analyze the pharmacokinetic study of XiaoShuanTongLuo Effective Components Group.In addition,absorption,tissue distribution,metabolism and excretion of SAA were studied.Whereas,pharmacokinetics and metabolism of pinocembrin were also studied in this dissertation.Partâ… XiaoShuanTongLuo Effective Components Group was studied in this dissertation.A sensitive method was developed,which was further applied to assess the pharmacokinetics of rutin,hyperoside,quercetin,kaempferol in rats after oral administration of XiaoShuanTongLuo Effective Components Group.After single oral doses of 300 mg/kg to rats,rutin,hyperoside,quercetin and kaempferol all appeared in rat plasma at 5min after administration.The peak plasma concentration occurred at 0.5,0.5,4,4 h with the C_max12.16±1.87,13.84±4.29,17.34±3.42,5.58±1.12 ng/ml,respectively.The AUC_0→24h were 72.59±14.23,99.01±16.62,161.60±32.83,60.31±12.88 ng·h/ml. Partâ…¡Pharmacokinetics and metabolism of SAA were studied,including absorption,tissue distribution,metabolism and excretion in rats.A sensitive analytical method was established for SAA in rat plasma,which was further applied to assess the pharmacokinetics and tissue distribution of SAA in rats receiving a single oral dose of SAA.Using the multi-stage MS(MSⁿ) method to analyze SAA and its metabolite of urine and bile in rats,the characteristic fragment ions were obtained.LC/MSⁿ method was eatablished to identify five metabolites.The effect of SAA on P450 enzymes in rat liver microsomes was evaluated for the first time.These studies provided scientific data for the new drug development and clinic use.1.Absorption pharmacokinetics in ratsTo investigate the absorption pharmacokinetics of SAA in rats,a sensitive analytical method was firstly established and validated for SAA in rat plasma,which was further applied to assess the pharmacokinetics of SAA in rats receiving a single oral dose of SAA.Linear calibration curves were obtained over the range of 1.4-1000 ng/ml for SAA in rat plasma.The lower limit of quantification was 1.4 ng/ml.Plasma concentration data were analyzed after i.v.or i.g.adminstration of SAA.2.Tissue distribution in ratsA LC/MS/MS method was developed to determine SAA in rat tissues.The results showed that SAA was distributed widely after oral administration to rats and could be found in many tissues.3.Metabolism of SAA in ratsMetabolites in rat urine and bile were investigated.A total of five metabolites were found in urine and bile by HPLC with ion trap mass spectrometric detection.Using the multi-stage MS(MSⁿ) method analysis of SAA and its metabolite,the characteristic fragment ions were obtained.A LC/MSⁿ method was eatablished to identify five metabolites.There were three main possible metabolic pathways of SAA in rat.In the first pathway SAA was absorbed into the blood and conjugated with methyl group.In the second pathway SAA was absorbed into the blood and conjugated with glucuronic acid group.In the third pathway SAA was absorbed into the blood and conjugated with both glucuronic acid group and methyl group.4.Excretion of SAA in ratsSAA was excreted approximately 0.102%into the bile in rats after 12 h post-dose.In feces,0.0022%was recovered after 48 h post-dose.In urine,0.014%was recovered after 48 h post-dose in rats. 5.The effect of SAA on P450 enzymes in rat liver microsomesUsing the CYP-specific substrate,the effect of SAA on CYP450s(CYP1A2, CYP2C9,CYP2C19,CYP2D6,CYP2E1 and CYP3A4) was evaluated both in vivo and in vitro studies.The data showed SAA neither induced nor inhibited the activities of CYP450s in rat liver microsomes.It was also found that SAA had no significant effect on rat liver microsomal protein and CYP450 content.Partâ…¢Pharmacokinetics and metabolism of pinocembrin were studied in this part.A sensitive analytical method was established for pinocembrin in Beagle dog plasma, which was further applied to assess the pharmacokinetics of pinocembrin in Beagle dog receiving a single oral dose of pinocembrin.After single oral doses of 100 mg/kg to Beagle dog,pinocembrin appeared in Beagle dog plasma at 5min after administration. The peak plasma concentration occurred at 120 min with the C_max 4697.40 ng/ml.Using the multi-stage MS(MSⁿ) method to analyze the metabolite of urine and plasma in Beagle dogs,the characteristic fragment ions were obtained.LC/MSⁿ method was established to identify two metabolites.