Title Page
Contents
국문초록 7
Abstract 9
Abbreviation 11
I. Introduction 17
1. Compounds 17
2. Drug-drug interaction 19
3. Physiological-based pharmacokinetic modeling 20
II. Objectives 23
III. Material and Method 24
1. Materials 24
2. Assessment of determining in vitro CYP450 enzyme inhibition potentials. 24
A. Nicardipine CYP450 inhibition assay 24
B. Data anaylsis 26
3. Screening of liver microsomes and recombinant CYP450 metabolic stability 27
A. Saxagliptin Metabolic Stability 27
B. Nateglinide Metabolic Stability 27
C. Data analysis 28
4. Non-clinical drug-drug interaction studies 29
A. Animal experiments 29
B. Study design and Sample collection 29
C. Sample preparation 30
D. Pharmacokinetic analysis of nicardipine, Saxagliptin and Nateginide. 31
5. Compound analysis using HPLC and LC-MS/MS 32
A. HPLC analysis 32
B. LC-MS/MS analysis 33
6. Physicochemical based pharmacokinetic (PBPK) modeling 34
A. Software and Data collection 34
B. The relative gene expression of enzyme and transporter 35
C. PBPK model construction and simulation 35
D. Model evaluation 36
IV. Results 38
1. Determination of in vitro CYP enzyme inhibition potential in liver microsome 38
2. in vitro recombinant CYP450 and liver microsomal metabolic stability 40
3. in vivo pharmacokinetic studies 43
4. in silico PBPK modeling and simulation 50
A. PBPK model structure 50
B. Model Validation 51
C. Prediction of drug-drug interaction for Rat and Human 58
V. Discussion 63
VI. Conclusion 68
VII. Reference 69
Appendices 78
Appendix 1-1. Nicardipine parameters for PBPK modeling 78
Appendix 1-2. Nicardipine parameter for PBPK modeling 79
Appendix 2-1. Saxagliptin parameter for PBPK modeling 80
Appendix 2-2. Saxagliptin parameter for PBPK modeling 81
Appendix 3-1. Nateglinide parameter for PBPK modeling 82
Appendix 3-2. Nateglinide parameter for PBPK modeling 83
Appendix 4. MRD values of plasma concentration prediction and GMFE values of AUCinf and Cmax prediction for nicardipine, saxagliptin and nateglinide[이미지참조] 84
Table 1. Mean (± Standard Deviation) pharmacokinetic parameters of nicardipine after oral administration 15mg/kg to rat 47
Table 2. Mean (± Standard Deviation) pharmacokinetic parameters of saxagliptin after oral single administration 5mg/kg and co-administration with nicardipine 15mg/kg to rat 48
Table 3. Mean (± standard deviation) pharmacokinetic parameters of nateglinide after oral single administration 20mg/kg and co-administration with nicardipine 15mg/kg to rat 49
Table 4. Comparison of observed and predicted pharmacokinetic parameters of nicardipine 55
Table 5. Comparison of observed and predicted pharmacokinetic parameters of saxagliptin 56
Table 6. Comparison of observed and predicted pharmacokinetic parameters of nateglinide 57
Table 7. Comparison of pharmacokinetic parameters for drug-drug interaction between saxagliptin and nicardipine 61
Table 8. Comparison of pharmacokinetic parameters for drug-drug interaction between nateglinide and nicardipine 62
Figure 1. Implemented drug-drug interaction process for saxagliptin, nateglinide and nicardipine. CYP : cytochrome P450 21
Figure 2. Drug-drug interaction structural overview of the PBPK modeling 22
Figure 3. Enzyme inhibition of CYP3A2 (A), CYP3A4 (B), CYP2C11 (C), and CYP2C9 (D) by nicardipine. All values were expressed as mean ±... 39
Figure 4. The metabolic stability of saxagliptin to derive metabolic half-life in rat liver microsome (A), human liver microsome (B), recombinant... 41
Figure 5. The metabolic stability of nateglinide to derive metabolic half-life in rat liver microsome (A), human liver microsome (B),... 42
Figure 6. The profile of mean plasma concentration - time curve of nicardipine in rats (n=7) following the oral administration 15mg/kg.... 44
Figure 7. The profile of mean plasma concentration - time curve of saxagliptin in rats following the oral administration (5mg/kg) and... 45
Figure 8. The profile of mean plasma concentration - time curve of nateglinide in rats following the oral administration (20mg/kg) and... 46
Figure 9. The semi-logarithmic scale plots and linear scale plot of plasma concentration - time profiles of nicardipine. Observed values... 52
Figure 10. The semi-logarithmic scale plots and of plasma concentration - time profiles of saxagliptin. Observed values from... 53
Figure 11. The semi-logarithmic scale plots and linear scale plot of plasma concentration - time profiles of nateglinide. Observed values... 54
Figure 12. The linear scale plot of plasma concentration - time profiles of saxaglipiin and nicardipine. The observed values in Figure 12-A were... 59
Figure 13. The linear scale plot of plasma concentration - time profiles of nateglinide and nicardipine. The observed values in Figure... 60
Figure 14. The profile of observed and predicted plasma concentration of nicardipine, saxagliptin and nateglinide for PBPK model. Each color... 66
Figure 15. Comparison to observed and predicted parameters (AUCinf and Cmax) of nicardipine, saxagliptin and nateglinide for PBPK model. Each color symbol represents compounds. Line of identity are marked the solid line....[이미지참조] 67