Title Page

Contents

List of Abbreviations 26

Abstract 29

Chapter 1. Introduction. 31

1.1. Permeability glycoprotein 31

1.2. Paclitaxel 32

1.3. Phytochemicals 34

1.3.1. Capsaicin 34

1.3.2. Resveratrol 35

1.3.3. Piperine 35

1.3.4. [6]-Gingerol 36

Chapter 2. Rationale and Aims of the Research 41

2.1. Rationale 41

2.2. Aims of the Research 42

Chapter 3. Materials and Methods 43

3.1. Materials 43

3.1.1. Reagents and Chemicals 43

3.1.2. Instruments 44

3.2. Analytical Methods for the Quantification of Paclitaxel in Plasma and Tissue Samples 45

3.2.1. HPLC/UV Detection Assay 45

3.3. Development Analytical Methods for the Quantification of Phytochemicals in Mice Plasma 49

3.3.1. LC/MS/MS Assay 49

3.4. Pharmacokinetic Studies of Phytochemicals in Mice 55

3.4.1. Pharmacokinetics of Capsaicin, resveratrol, Piperine and [6]-Gingerol after Intravenous Injection 55

3.4.2. Pharmacokinetics of Capsaicin, Resveratrol, Piperine and [6]-Gingerol after Intraperitoneal Injection. 56

3.4.3. Effects of Capsaicin and Resveratrol on the Plasma Pharmacokinetics and Tissue Distribution of Paclitaxel 56

3.4.4. Data Analysis 60

3.4.5. Statistical Analysis 60

Chapter 4. Results 61

4.1. Analytical Methods for the Quantification of Paclitaxel in Plasma and Tissue Samples 61

4.1.1. HPLC/UV Detection Assay 61

4.2. Analytical Methods for the Quantification of Phytochemicals in Mice Plasma 68

4.2.1. Mass Spectrometry and Chromatography 68

4.2.2. Validation 84

4.3. Pharmacokinetic Studies of Phytochemical in Mice 101

4.3.1. Pharmacokinetics of Capsaicin after Intravenous and Intraperitoneal Injection 101

4.3.2. Pharmacokinetics of Resveratrol after Intravenous and Intraperitoneal Injection 114

4.3.3. Pharmacokinetics of Piperine after Intravenous and Intraperitoneal Injection 127

4.3.4. Pharmacokinetics of [6]-Gingerol after Intravenous and Intraperitoneal Injection 140

4.4. Effects of Phytochemicals on the Plasma Pharmacokinetics and Tissue Distribution of Paclitaxel in Mice 153

4.4.1. Animal Pretreatment with Capsaicin and Resveratrol and Attainment of Steady State 153

4.4.2. Effects of Phytochemicals on the Plasma Pharmacokinetics of Paclitaxel in Mice 155

4.4.3. Effects of Phytochemicals on the Tissue Distribution of Paclitaxel in Mice 181

Chapter 5. Discussion 193

5.1. Analytical Methods for the Quantification of Paclitaxel in Plasma and Tissue Samples 193

5.1.1. HPLC/UV Detection Assay 193

5.2. Analytical Methods for the Quantification of Phytochemicals in Mice Plasma 194

5.2.1. LC/MS/MS Assay 194

5.3. Pharmacokinetic Studies of Phytochemicals in Mice 196

5.3.1. Pharmacokinetics of Capsaicin after Intravenous and Intraperitoneal Injection 196

5.3.2. Pharmacokinetics of Resveratrol after Intravenous and Intraperitoneal Injection 196

5.3.3. Pharmacokinetics of Piperine after Intravenous and Intraperitoneal Injection 197

5.3.4. Pharmacokinetics of [6]-Gingerol after Intravenous and Intraperitoneal Injection 198

5.4. Effects of Phytochemicals on the Plasma Pharmacokinetics and Tissue Distribution of Paclitaxel in Mice 199

5.4.1. Animal Pretreatment with Capsaicin and Resveratrol and Attainment of Steady State 199

5.4.2. Effects of Phytochemicals on the Plasma Pharmacokinetics of Paclitaxel in Mice 200

5.4.3. Effects of Phytochemicals on the Tissue distribution of Paclitaxel in Mice 202

Chapter 6. Conclusion 204

Chapter 7. References 206

국문요약 216

Table 4-1. Linearity of paclitaxel assay in mice plasma and various tissues 66

Table 4-2. Average intra- and inter-day accuracy and precision of paclitaxel assay in mice plasma and various tissues (n ＝ 5 each) 67

Table 4-3. Summary of calibration curves used in method validation for capsaicin 90

Table 4-4. Summary of calibration curves used in method validation for resveratrol 91

Table 4-5. Summary of calibration curves used in method validation for piperine 92

Table 4-6. Summary of calibration curves used in method validation for [6]-gingerol 93

Table 4-7. Average intra- and inter-day accuracy (mean ± S.D.) and precision of capsaicin in mice plasma (n ＝ 5 each) 94

Table 4-8. Average intra- and inter-day accuracy (mean ± S.D.) and precision of resveratrol in mice plasma (n ＝ 5 each) 95

Table 4-9. Average intra- and inter-day accuracy (mean ± S.D.) and precision of pipreine in mice plasma (n ＝ 5 each) 96

Table 4-10. Average intra- and inter-day accuracy (mean ± S.D.) and precision of [6]-gingerol in mice plasma (n ＝ 5 each) 97

Table 4-11. Stability of capsaicin 98

Table 4-12. Stability of [6]-gingerol 99

Table 4-13. Average process efficiency of [6]-gingerol (analyte) and nonivamide (internal standard, IS) (n ＝ 3 each, mean ± S.D.) 100

Table 4-14. Average plasma concentration (mean ± S.D.) of capsaicin (ng/mL) vs. time data observed after i.v. injection (doses 0.325, 0.65mg/kg) and i.p. injection of capsaicin in mice (dose 5mg/kg) (n ＝ 4 - 5 each) 106

Table 4-15. Plasma capsaicin concentration (ng/mL) vs. time data observed after i.v. injection of capsaicin in mice (dose 0.325mg/kg) 107

Table 4-16. Plasma capsaicin concentration (ng/mL) vs. time data observed after i.v. injection of capsaicin in mice (dose 0.65mg/kg) 108

Table 4-17. Plasma capsaicin concentration (ng/mL) vs. time data observed after i.p. injection of capsaicin in mice (dose 5mg/kg) 109

Table 4-18. Average pharmacokinetic parameters (mean ± S.D.) of capsaicin obtained after i.v. injection (doses 0.325, 0.65mg/kg) and i.p injection (dose 5mg/kg) of capsaicin in mice (n ＝ 4 - 5 each) 110

Table 4-19. Pharmacokinetic parameters of capsaicin obtained after i.v. injection of capsaicin in mice (dose 0.325mg/kg) 111

Table 4-20. Pharmacokinetic parameters of capsaicin obtained after i.v. injection of capsaicin in mice (dose 0.65mg/kg) 112

Table 4-21. Pharmacokinetic parameters of capsaicin obtained after i.p. injection of capsaicin in mice (dose 5mg/kg) 113

Table 4-22. Average plasma concentration (mean ± S.D.) of resveratrol (ng/mL) vs. time data observed after i.v. injection (doses 3, 6mg/kg) and i.p. injection of resveratrol in mice (dose 5mg/kg) (n ＝ 4 - 5 each) 119

Table 4-23. Plasma resveratrol concentration (ng/mL) vs. time data observed after i.v. injection of resveratrol in mice (dose 3mg/kg) 120

Table 4-24. Plasma resveratrol concentration (ng/mL) vs. time data observed after i.v. injection of resveratrol in mice (dose 6mg/kg) 121

Table 4-25. Plasma resveratrol concentration (ng/mL) vs. time data observed after i.p. injection of resveratrol in mice (dose 5mg/kg) 122

Table 4-26. Average pharmacokinetic parameters (mean ± S.D.) of resveratrol obtained after i.v. injection (doses 3, 6mg/kg) and i.p. injection (dose 5mg/kg) of resveratrol in mice (n ＝ 4 - 5 each) 123

Table 4-27. Pharmacokinetic parameters of resveratrol obtained after i.v. injection of resveratrol in mice (dose 3mg/kg) 124

Table 4-28. Pharmacokinetic parameters of resveratrol obtained after i.v. injection of resveratrol in mice (dose 6mg/kg) 125

Table 4-29. Pharmacokinetic parameters of resveratrol obtained after i.p. injection of resveratrol in mice (dose 5mg/kg) 126

Table 4-30. Average plasma concentration (mean ± S.D.) of piperine (ng/mL) vs. time data observed after i.v. injection (doses 2, 4mg/kg) and i.p. injection of piperine in mice (dose 5mg/kg) (n ＝ 5 each) 132

Table 4-31. Plasma piperine concentration (ng/mL) vs. time data observed after i.v. injection of piperine in mice (dose 2mg/kg) 133

Table 4-32. Plasma piperine concentration (ng/mL) vs. time data observed after i.v. injection of piperine in mice (dose 4mg/kg) 134

Table 4-33. Plasma piperine concentration (ng/mL) vs. time data observed after i.p. injection of piperine in mice (dose 5mg/kg) 135

Table 4-34. Average pharmacokinetic parameters (mean ± S.D.) of piperine obtained after i.v. injection (doses 2, 4mg/kg) and i.p. injection (dose 5mg/kg) of piperine in mice (n ＝ 4 - 5 each) 136

Table 4-35. Pharmacokinetic parameters of piperine obtained after i.v. injection of piperine in mice (dose 2mg/kg) 137

Table 4-36. Pharmacokinetic parameters of piperine obtained after i.v. injection of piperine in mice (dose 4mg/kg) 138

Table 4-37. Pharmacokinetic parameters of piperine obtained after i.p. injection of piperine in mice (dose 5mg/kg) 139

Table 4-38. Average plasma concentration (mean ± S.D.) of [6]-gingerol (ng/mL) vs. time data observed after i.v. injection (doses 3, 6mg/kg) and i.p. injection of [6]-gingerol in mice (dose 5mg/kg) (n ＝ 4 - 5 each) 145

Table 4-39. Plasma [6]-gingerol concentration (ng/mL) vs. time data observed after i.v. injection of [6]-gingerol in mice (dose 3mg/kg) 146

Table 4-40. Plasma [6]-gingerol concentration (ng/mL) vs. time data observed after i.v. injection of [6]-gingerol in mice (dose 6mg/kg) 147

Table 4-41. Plasma [6]-gingerol concentration (ng/mL) vs. time data observed after i.p. injection of [6]-gingerol in mice (dose 5mg/kg) 148

Table 4-42. Average pharmacokinetic parameters (mean ± S.D.) of [6]-gingerol obtained after i.v. injection (doses 3, 6mg/kg) and i.p. injection (dose 5mg/kg) of [6]-gingerol in mice (n ＝ 4 - 5 each) 149

Table 4-43. Pharmacokinetic parameters of [6]-gingerol obtained after i.v. injection of [6]-gingerol in mice (dose 3mg/kg) 150

Table 4-44. Pharmacokinetic parameters of [6]-gingerol obtained after i.v. injection of [6]-gingerol in mice (dose 6mg/kg) 151

Table 4-45. Pharmacokinetic parameters of [6]-gingerol obtained after i.p. injection of [6]-gingerol in mice (dose 5mg/kg) 152

Table 4-46. Average plasma concentration (mean ± S.D.) of paclitaxel (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with control vehicle, capsaicin and resveratrol using Alzet osmotic pump (n ＝ 4 - 5 each) 160

Table 4-47. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with control vehicle using Alzet osmotic pump (dose 5mg/kg) 161

Table 4-48. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with capsaicin using Alzet osmotic pump (dose 5mg/kg) 162

Table 4-49. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with resveratrol using Alzet osmotic pump (dose 5mg/kg) 163

Table 4-50. Average pharmacokinetic parameters (mean ± S.D.) of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle, capsaicin and resveratrol using Alzet osmotic pump (n ＝ 4 - 5 each) (dose 5mg/kg) 164

Table 4-51. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle (dose 5mg/kg) 165

Table 4-52. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with capsaicin (dose 5mg/kg) 166

Table 4-53. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with resveratrol (dose 5mg/kg) 167

Table 4-54. Average plasma concentration of paclitaxel (mean ± S.D.) of paclitaxed (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with control vehicle, piperine and [6]-gingerol (n ＝ 4 - 5 each) 173

Table 4-55. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with control vehicle (dose 5mg/kg) 174

Table 4-56. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with piperine (dose 5mg/kg) 175

Table 4-57. Plasma paclitaxel concentration (ng/mL) vs. time data observed after i.v. injection of paclitaxel in mice pretreated with [6]-gingerol (dose 5mg/kg) 176

Table 4-58. Average pharmacokinetic parameters (mean ± S.D.) of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle, piperine and [6]-gingerol (n ＝ 4 - 5 each) (dose 5mg/kg) 177

Table 4-59. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle (dose 5mg/kg) 178

Table 4-60. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with piperine (dose 5mg/kg) 179

Table 4-61. Pharmacokinetic parameters of paclitaxel obtained after i.v. injection of paclitaxel in mice pretreated with [6]-gingerol (dose 5mg/kg) 180

Table 4-62. Average paclitaxel concentrations (ng/g, mean ± S.D.)in tissues after i.v. injection of paclitaxel (dose 5mg/kg) in mice pretreated with control vehicle, capsaicin and resveratrol 184

Table 4-63. Fold increase of tissues paclitaxel concentrations in mice pretreated with capsaicin and resveratrol as compared with control vehicle 185

Table 4-64. Tissue-to-plasma partition coefficients (Kp) of paclitaxel in mice pretreated with control vehicle, capsaicin and resveratrol (mean ± S.D.)(이미지참조) 186

Table 4-65. Average paclitaxel concentrations (mean ± S.D.) in tissues after i.v. injection of paclitaxel (dose 5mg/kg) in mice pretreated with control vehicle, piperine and [6]-gingerol 190

Table 4-66. Fold increase of tissues paclitaxel concentrations in mice pretreated with piperine and [6]-gingerol compared with control vehicle 191

Table 4-67. Tissue-to-plasma partition coefficients (Kp) of paclitaxel in mice pretreated with control vehicle, piperine and [6]-gingerol (mean ± S.D.)(이미지참조) 192

Figure 1-1. Structure of paclitaxel. 33

Figure 1-2. Structure of capsaicin. 37

Figure 1-3. Structure of resveratrol. 38

Figure 1-4. Structure of piperine. 39

Figure 1-5. Structure of [6]-gingerol. 40

Figure 4-1. Chormatograms of paclitaxel (A) and the internal standard (IS) (B) obtained by extraction of a mouse plasma spike with paclitaxel (10000ng/mL) and the internal standard (2500ng/mL) (paclitaxel rt ＝ 7.8 min, internal standard rt ＝ 6.4 min).(이미지참조) 63

Figure 4-2. Chormatograms of paclitaxel (A) and the internal standard (IS) (B) obtained by extraction of a mouse plasma obtained 1 hr after oral administration of paclitaxel to mice at a dose of 40mg/kg. (paclitaxel rt ＝ 7.8 min, internal standard rt ＝ 6.4 min).(이미지참조) 64

Figure 4-3. A representative calibration curve for the determination of paclitaxel in plasma (y ＝ 0.001266x - 0.0041, r² ＝ 0.9979). 65

Figure 4-4. (A) Positive ion Q1 mass spectrum of capsaicin and (B) product ion mass of protonated capsaicin ([M+H]+, m/z 306.4).(이미지참조) 69

Figure 4-5. (A) Positive ion Q1 mass spectrum of phenacetin and (B) product ion mass of protonated phenacetin ([M+H]+, m/z 180.1).(이미지참조) 70

Figure 4-6. MRM chromatograms of capsaicin (left) and phenacetin (right) obtained by extraction of (A) blank mouse plasma, (B) plasma spiked with capsaicin (0.325ng/mL, LLOQ) and phenacetin (50ng/mL), (C) plasma sample obtained 30 min after intravenous injection of capsaicin to a mouse (dose 0.65mg/kg). 71

Figure 4-7. (A) Negative ion Q1 mass spectrum of resveratrol and (B) product ion mass of deprotonated resveratrol ([M-H]-, m/z 227.3).(이미지참조) 73

Figure 4-8. (A) Negative ion Q1 mass spectrum of 4,5,7 trihydroxyflavanone and (B) product ion mass of deprotonated 4,5,7 trihydroxyflavanone([M-H]-, m/z 271.3).(이미지참조) 74

Figure 4-9. MRM chromatograms of resveratrol (left) and 4,5,7 trihydroxyflavanone (right) obtained by extraction of (A) blank mouse plasma, (B) plasma spiked with resveratrol (500ng/mL) and 4,5,7 trihydroxyflavanone (500ng/mL), (C) plasma sample obtained 5 min after intravenous injection of resveratrol to a mouse (dose 3mg/kg). 75

Figure 4-10. (A) Positive ion Q1 mass spectrum of piperine and (B) product ion mass of protonated piperine ([M+H]+, m/z 286.3).(이미지참조) 77

Figure 4-11. (A) Positive ion Q1 mass spectrum of phenacetin and (B) product ion mass of protonated phenacetin ([M+H]+, m/z 180.1).(이미지참조) 78

Figure 4-12. MRM chromatograms of piperine (left) and phenacetin (right) obtained by extraction of (A) blank mouse plasma, (B) plasma spiked with piperine (100ng/mL) and phenacetin (50ng/mL), (C) plasma sample obtained 30 min after intravenous injection of piperine to a mouse (dose 4mg/kg). 79

Figure 4-13. (A) Positive ion Q1 mass spectrum of [6]-gingerol and (B) product ion mass of dehydrated [6]-gingerol ([M+H-H2O]+, m/z 277.2).(이미지참조) 81

Figure 4-14. (A) Positive ion Q1 mass spectrum of nonivamide and (B) product ion mass of protonated nonivamide ([M+H]+, m/z 294.2).(이미지참조) 82

Figure 4-15. MRM chromatograms of [6]-gingerol (left) and nonivamide (right) obtained by extraction of (A) blank mouse plasma, (B) plasma spiked with [6]-gingerol (500ng/mL) and nonivamide (500mg/mL), (C) plasma sample obtained 4 hr after intravenous injection of [6]-gingerol to a mouse (dose 3mg/kg). 83

Figure 4-16. A representative calibration curve for the determination of capsaicin in plasma (y ＝ 0.0034x - 0.000342, r ＝ 0.9997). 86

Figure 4-17. A representative calibration curve for the determination of resveratrol in plasma (y ＝ 0.000497x - 0.00021, r ＝ 0.9999). 87

Figure 4-18. A representative calibration curve for the determination of piperine in plasma (y ＝ 0.0195x + 0.00753, r ＝ 0.9997). 88

Figure 4-19. A representative calibration curve for the determination of [6]-gingerol in plasma (y ＝ 0.000112x + 0.0000941, r ＝ 0.9996). 89

Figure 4-20. Average plasma capsaicin concentration (mean ± S.D.) vs. time curves obtained after i.v. injection (doses 0.325, 0.65mg/kg) and i.p. injection (5mg/kg) of capsaicin in mice. 102

Figure 4-21. Plasma capsaicin concentration vs. time curves obtained after i.v. injection of capsaicin in mice (dose 0.325mg/kg). 103

Figure 4-22. Plasma capsaicin concentration vs. time curves obtained after i.v. injection of capsaicin in mice (dose 0.65mg/kg). 104

Figure 4-23. Plasma capsaicin concentration vs. time curves obtained after i.p. injection of capsaicin in mice (dose 5mg/kg). 105

Figure 4-24. Average plasma resveratrol concentration (mean ± S.D.) vs. time curves obtained after i.v. injection (doses 3, 6mg/kg) and i.p. injection (5mg/kg) of resveratrol in mice. 115

Figure 4-25. Plasma resveratrol concentration vs. time curves obtained after i.v. injection of resveratrol in mice (dose 3mg/kg). 116

Figure 4-26. Plasma resveratrol concentration vs. time curves obtained after i.v. injection of resveratrol in mice (dose 6mg/kg). 117

Figure 4-27. Plasma resveratrol concentration vs. time curves obtained after i.p. injection of resveratrol in mice (dose 5mg/kg). 118

Figure 4-28. Average plasma piperine concentration (mean ± S.D.) vs. time curves obtained after i.v. injection (doses 2, 4mg/kg) and i.p. injection (5mg/kg) of piperine in mice. 128

Figure 4-29. Plasma piperine concentration vs. time curves obtained after i.v. injection of piperine in mice (dose 2mg/kg). 129

Figure 4-30. Plasma piperine concentration vs. time curves obtained after i.v. injection of piperine in mice (dose 4mg/kg). 130

Figure 4-31. Plasma piperine concentration vs. time curves obtained after i.p. injection of piperine in mice (dose 5mg/kg). 131

Figure 4-32. Average plasma [6]-gingerol concentration (mean ± S.D.) vs. time curves obtained after i.v. injection (doses 3, 6mg/kg) and i.p. injection (5mg/kg) of [6]-gingerol in mice. 141

Figure 4-33. Plasma [6]-gingerol concentration vs. time curves obtained after i.v. injection of [6]-gingerol in mice (dose 3mg/kg). 142

Figure 4-34. Plasma [6]-gingerol concentration vs. time curves obtained after i.v. injection of [6]-gingerol in mice (dose 6mg/kg). 143

Figure 4-35. Plasma [6]-gingerol concentration vs. time curves obtained after i.p. injection of [6]-gingerol in mice (dose 5mg/kg). 144

Figure 4-36. Average plasma concentration-time profiles of capsaicin and resveratrol in mice during i.p. infusion to steady-state using Alzet osmotic pump. (mean ± S.D.). 154

Figure 4-37. Average plasma paclitaxel concentration (mean ± S.D.) vs. time profiles after i.v. injection in mice pretreated with control vehicle, capsaicin and resveratrol using Alzet osmotic pump (n ＝ 5 each, paclitaxel dose 5mg/kg). 156

Figure 4-38. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle (dose 5mg/kg). 157

Figure 4-39. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with caspsaicin (dose 5mg/kg). 158

Figure 4-40. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with resveratrol (dose 5mg/kg). 159

Figure 4-41. Average plasma paclitaxel concentration (mean ± S.D.) vs. time profiles after i.v. injection in mice pretreated with control vehicle, piperine and [6]-gingerol (n ＝5 each, paclitaxel dose 5mg/kg). 169

Figure 4-42. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with control vehicle (dose 5mg/kg). 170

Figure 4-43. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with piperine (dose 5mg/kg). 171

Figure 4-44. Plasma paclitaxel concentration vs. time curves obtained after i.v. injection of paclitaxel in mice pretreated with [6]-gingerol (dose 5mg/kg). 172

Figure 4-45. Paclitaxel concentrations (mean ± S.D.) in tissues after i.v. injection of paclitaxel (dose 5mg/kg) in mice pretreated with control vehicle (□, n ＝ 5), capsaicin (■, n ＝ 4 - 5), and resveratrol (▨, n ＝ 5). 183

Figure 4-46. Paclitaxel concentrations (mean ± S.D.) in tissues after i.v. injection of paclitaxel (dose 5mg/kg) in mice pretreated with control vehicle (□, n ＝ 5), piperine (■, n ＝ 5), and [6]-gingerol (▨, n ＝ 4 - 5). 189

본 연구에서는 천연물 유래 P-gp 억제제인 capsaicin, resveratrol, piperine 및 [6]-gingerol의 마우스에서의 체내동태를 규명하고 이 물질들이 P-gp 기질 약물인 paclitaxel의 혈중 약물동태와 조직 분포에 미치는 영향을 규명하고자 하였다. 생체시료 중 각 P-gp 억제제의 농도 정량은 LC/MS/MS 방법을 개발하여 사용하였고 paclitaxel의 정량은 HPLC/UV 방법을 적용하였다. 개발된 LC/MS/MS의 정량한계는 20 μL의 생체시료를 사용하였을 때 capsaicin, resveratrol, piperine 및 [6]-gingerol의 경우 각각 0.325, 10, 0.5, 및 10ng/mL 이었다. Paclitaxel의 정량한계는 20 μL의 생체시료를 사용하였을 때 50ng/mL 이었다. 각 P-gp 억제제의 마우스에서의 체내동태를 정맥 및 복강 투여한 후 채혈을 통하여 규명하였다. Capsaicin, resveratrol 및 [6]-gingerol을 복강투여 하였을 때 거의 온전히 흡수되어 절대 절대생체이용률은 각각 82.3 - 125.7%, 81.4 - 114.5%, 104.1 - 153.1%이었으나, piperine은 흡수율이 낮았다 (절대생체이용률 15.5 - 18.8%). P-gp 억제제가 pcalitaxel의 체내동태와 조직 분포에 미치는 영향을 확인하기 위하여 마우스를 capsaicin, resveratrol, piperine 또는 [6]-gingerol로 전처리하였다. Capsaicin과 resveratrol의 경우, 각 물질을 마우스에 Alzet osmotic pump를 이용하여 등속복강주입하고 정상상태 (steady-state)에 도달하였을 때 기질 약물인 paclitaxel을 단회 정맥주사하고 혈액, 위, 소장, 대장, 간, 고환, 심장, 신장, 비장, 폐 그리고 뇌 시료를 채취하였다. Piperine과 [6]-ginerol의 경우, 각 물질을 마우스에 단회 복강 주사 후 paclitaxel을 즉시 단회 정맥주사하고 시료를 채취하였다. Capsaicin과 piperine으로 전처리한 마우스 군에서 혈중 농도 곡선 하 면적 (AUC)과 전신클리어런스 (CLs)가 대조군에 비해 각각 유의성 있게 증가 및 감소하였다. 또한, capsaicin, piperine 및 [6]-gingerol로 전처리한 마우스 군에서 paclitaxel의 조직 분포가 대조군에 비해 유의성 있게 증가하였다. 요약하면, 본 연구에서는 마우스에서의 천연물 유래 P-gp 억제제인 capsaicin, resveratrol, piperine과 [6]-gingerol의 체내동태를 규명하였고 각 P-gp 억제제가 paclitaxel의 혈중 동태와 조직 분포에 미치는 영향을 규명하였다. capsaicin과 piperine이 paclitaxel의 혈중 체내동태에 유의성 있는 영향을 나타내었으며, capsaicin, piperine 및 [6]-gingerol이 paclitaxel의 조직 분포에 유의성있는 영향을 나타내었다.