Title Page
Contents
GLOSSARY OF TERMS 11
I. Introduction 13
1. Corona Virus Disease 19 (COVID-19) 13
2. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 14
3. CNC1049 14
4. Tandem Mass Spectrometry (MS/MS) 15
1) MS/MS 15
2) Electrospray Ionization (ESI) 16
5. Purpose 16
II. Materials and Methods 18
1. Chemicals and Solvents 18
2. Equipment 18
1) Equipment used to collect biological samples 18
2) Equipment used for preparation of biological samples 19
3) Instrument 20
4) Animals 20
3. Analytical procedures of CNC1049 in biological samples 22
1) Stock solution 22
2) Plasma and tissue sample preparation 22
3) Urine and feces sample preparation 23
4) HPLC-MS/MS analysis condition 24
4. Analytical method validation 29
1) Specificity 29
2) Linearity 29
3) Precision and Accuracy 29
4) Stability 30
5) Matrix effect, recovery, and process efficiency 31
5. Pharmacokinetic study 32
1) In vitro study 32
2) In vivo study in rats 38
3) Pharmacokinetic parameters analysis 42
4) Statistical analysis 43
III. Results 44
1. Analytical method validation 44
1) Specificity 44
2) Linearity 44
3) Precision and Accuracy 47
4) Stability 47
5) Matrix effect, Recovery, and Process efficiency 47
2. Pharmacokinetics study 51
1) In vitro study 51
2) In vivo study in rats 59
IV. Discussion and Conclusion 78
V. References 84
Abstract 87
Table 1. Analytical condition of HPLC (Agilent 1100 series) for CNC1049 and... 27
Table 2. Analytical condition of MS/MS (API-000 QTrap) for CNC1049 and... 28
Table 3. Information of rat liver microsome 33
Table 4. Information of enzyme, substrate and inhibitor for CYP isozyme 35
Table 5. Intra- and inter-day precision and accuracy of CNC1049 in rat plasma 48
Table 6. Stability of CNC1049 in rat plasma under various conditions 49
Table 7. Matrix effect, recovery, and process efficiency of CNC1049 in rat plasma. 50
Table 8. Microsomal stability of CNC1049 and reference 52
Table 9. CYP inhibition by CNC1049 54
Table 10. Plasma protein binding in mice, rat, dog, monkey and human 58
Table 11. Plasma concentration of CNC1049 after a single intravenous injection in fasted SD rats 61
Table 12. Pharmacokinetic parameters of CNC1049 after a single intravenous injection in fasted SD rats 62
Table 13. Plasma concentration of 1 mg/kg CNC1049 after a single intravenous injection, oral administration and intra-... 65
Table 14. Pharmacokinetic parameters of 1 mg/kg CNC1049 after a single intravenous injection, oral administration and... 66
Table 15. Urinary and fecal excretion of CNC1049 after 3 mg/kg intravenous... 68
Table 16. Tissues concentration after oral administration of 300 mg/kg... 75
Table 17. Maximum concentration (Cmax) and time of Cmax (Tmax) of CNC1049...[이미지참조] 76
Table 18. Tissue to plasma partition coefficient (Kp) after oral administration of...[이미지참조] 77
Figure 1. Chemical structures of (a) CNC1049 and (b) Ibuprofen (IS) 21
Figure 2. Production mass spectra of (a) CNC1049 and (b) Ibuprofen (IS) 25
Figure 3. Principle of plasma protein binding assay 37
Figure 4. HPLC-MS/MS chromatograms of CNC1049 and IS in rat plasma.... 45
Figure 5. Calibration curve of CNC1049 in rat plasma 46
Figure 6. Sigmoidal curve of CYP2C9 activity suppressed by CNC1049. Data... 56
Figure 7. Plasma concentration-time curves of CNC1049 after single intrave-... 60
Figure 8. Plasma concentration-time curves of 1 mg/kg CNC1049 after intrave-... 64
Figure 9. Urinary excreted amount-time curve of CNC1049 after 3 mg/kg intra-... 69
Figure 10. Fecal excreted amount-time curve of CNC1049 after 3 mg/kg intra-... 70
Figure 11. Tissue concentration compared with plasma concentration of... 72
Figure 12. Tissue to plasma partition coefficient (Kp) of CNC1049 after oral administration in fasted SD rats[이미지참조] 74