| Background and objective: most of Anti-HBV medicine widely used in clinic belong to Nucleotide compound. Such drugs'resistance and end-of-dose failure become more and more evident. GLSBr, synthesized chemically, is the non-nucleoside compound, which has showed a new mechanism of anti-HBV and a significant effect of anti-HBV in animal experiments both in vitro and vivo. However, At least 40% of the new compounds have been eliminated because their absorption, distribution, metabolism and excretion are not ideal. Therefore, studying the pharmacodynamic features and its absorptive mechanism is playing a decisive role in making a new anti-HBV medicine. To examine and analyze the pharmacokinetic features of GLSBr series and study the absorptive mechanisms of the advantageous compound are the purpose of this thesis, which will provide a theoretical and examining foundation for the research of fresh anti-HBV medicine and selecting the advantageous GLSBr compounds.Method:1. First of all, to explore and establish a RP-HPLC method of detecting biologic samples and to make a systematic inspection of it. 48 SD rats are selected and divided into 12 groups randomly. The gavage or intravenous doses were divided into two levels of GLSBr-1, GLSBr-2 and GLSBr-3 solution. Next adopt a series of blood samples from the vein before and after the administration, and then put them into tubes with anticoagulant heparin. Finally, to analyze and compare the pharmacokinetics characteristics and differences respectively by using software DAS2.0 to calculate the major parameters of GLSBr compounds.2. Select another 24 SD big rats, divide randomly them into 6 groups and give them respectively mainlines (GLSBr-1,GLSBr-2 and GLSBr-3) though their tails . In each group, the heads of 4 rats are cut after the drug given in 10 minutes and the same way done to the heads of another 4 rats but in 2 hours. Next, extract the blood samples, livers, kidneys and brain organs and then detecting concentration of them by RP-HPLC.3. Choose LC-MS-MS to make a qualitative analysis of the suspected GLSBr-3'metabolites tested by RP-HPLC. Then speculate and set up the LC-MS checking methods of GLSBr-3 archetype and metabolized substance to make a qualitative analysis of interior functional process. This tissue distribution feature is favor of educing effect of GLSBr-1,GLSBr-2 in livers.4. Build up and make a full evaluation of the features of Caco-2 monolayer cell models, which can be adopted to observe the absorptive process of highly effective GLSBr-3. Review the influence of time, bottom concentration, system temperature, cultivated medium PH value and P-gp depressor Ver on the ingestion of GLSBr-3 from Caco-2 cells. Determine the concentration of GLSBr-3 inside and outside Caco-2 cells via the RP-HPLC checking methodology. And use SPSS12.0, Excel 2003 software analysis and statistical processing of data from the trial.Results:1. The established RP-HPLC/LC-MS detection method of the plasma samples and tissue samples has high sensitivity and specificity, and methods of inspection fits to the requirements of biological samples.2. GLSBr-1 and GLSBr-2's physiological disposition fit to the two-compartment model. When the GLSBr-1 is 80 mg / kg and GLSBr-2 is 11mg/kg , they accord with the first-order kinetics . GLSBr-1 and GLSBr-2 of the t1/2Zin the rat were 4.56±1.67h and 0.95±1.21h; and GLSBr-1 in the body at the time is longer than GLSBr-2 (P <0.05); GLSBr-3 in the RP-HPLC conditions can not be detected .3. Intravenous injection of different doses of the GLSBr-1 and GLSBr-2 ,the highest measured concentration is 1.25±0.23 and 0.61±0.27g·mL-1 in the liver; and in kidney concentration is next: 0.51±0.37 and 0.26±0.08 g·mL-1; in brain the GLSBr-1 's concentration is 0.34±0.10μg·mL-1, and GLSBr-2 can not be measured. Distribution of such an organization has enhanced the role-playing of GLSBr-1 and GLSBr-2 drugs in the liver.4. In rat plasma, GLSBr-3' chromatographic peak has not been detected by RP-HPLC, but a suspicious metabolism which is closely related to concentration stably appears. Using the LC-MS-MS analysis and testing, the metabolin of GLSBr-3 is from C21H22N4O3FSBr to C19H18N4O3FSBr by ester hydrolysis. LC-MS test results showed that although in mice plasma the metabolin concentrations is higher than the prototype drug concentration; in liver and kidney, the GLSBr-3 concentration is significantly higher than its metabolin. The highest measured concentration is in the liver, which is about 0.79±0.08μg·mL-1 . And its duration can be up to above 8 h, which shows it has an advantage of the liver distributing tendency.5. Caco-2 cell model experiments showed that the former GLSBr-3 increased with the time. Within the concentration of 6.25 50μg / mL , its intake showed a linear increase with the increase in the concentration , there is no saturated phenomenon. However , the temperature has a significant effect on the GLSBr-3 intake (P <0.05 ) with a PDR value of 1.335±0.141. It indicates that there are two ways of intake when Caco-2 cells is incepting GLSBr-3 : passive diffusion and active transport. And GLSBr-3 may be non-P-gp because the special features of P-gp inhibit verapamil from affecting the intake of GLSBr-3.Conclusion:The established RP-HPLC/LC-MS detection method is stable and can be used to study the test. GLSBr-1 and GLSBr-2 in rats fit pharmacokinetics of the two-model features. The distributing features of a series of GLSBr compounds in rats are: liver> kidney> brain, which can enhance the role of liver playing in the anti-HBV process. Although GLSBr-3 in the body quickly generated metabolin C19H18N4O3FSBr by ester hydrolysis, GLSBr-3' concentration in liver is significantly higher than its metabolin, and can stay longer. Based on the GLSBr-3' anti-HBV effect and distribution of liver trend, the GLSBr-3 may become the most potential compound. GLSBr-3 has the passive and active co-existence of the rotation transfer mode.
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