結論與建議

一、 Genipin 之 pKa≒6.3。其原形分子於血清中極不安定。

二、大鼠靜脈注射 genipin 後，立即代謝成 sulfate 結合態代謝物，其原形分子只短暫存在於血漿中。

三、大鼠口服 genipin 後，主要以 genipin sulfate 存在於血循環中，顯示此代謝物於體內發揮藥理作用之重要性值得探究。口服 200 mg/kg genipin 的大鼠死亡率高達 77.8%。

四、大鼠口服梔子水煎劑後，主要以 genipin sulfate 循環於體內，滯留體內時間達六天之久，且有明顯的腸肝循環現象，而 genipin 與 genipin glucuronide 於血中無法偵測。

五、併服梔子水煎劑對 MTX 之 AUC、 MRT 顯著增加，CL/F 顯著下降，顯示梔子水煎劑抑制 MTX 之排除，並造成部份大鼠死亡。

六、併服梔子水煎劑顯著減少 PHT 給藥後之早期暴露及 CBZ 之全期暴露。

七、建議使用 MTX、PHT 及 CBZ 之患者盡量避免併服梔子水煎劑及其相關方劑，以免造成中西藥交互作用，以確保療效及用藥安全。

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Volumn of NaOH (mL)

Fig. 1 The titration curve of genipin.

Volumn of NaOH (mL)

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4

Fig. 2 The first derivative (△pH/△V) titration curve of genipin.

Volumn of NaOH (mL)

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4

Time (h)

0 20 40 60 80 100 120 140 160

Remaining percentage (%) of genipin ⁶⁰

70 80 90 100

Fig. 4 Stability of genipin in MeOH (n=3).

A B C

Remaining percentage (%) of genipin

0 20 40 60 80 100

Fig. 5 Stability of genipin in serum stored at -20℃ (n=3).

(A) processed immediately (B) stored at -20℃for 1 day (C) stored at -20℃for 3 days

***P＜0.001

***

A B C

Remaining percentage (%) of genipin

0 20 40 60 80 100

Fig. 6 Effect of ascorbic acid on genipin stability in serum (n=3).

(A) processed immediately

(B) stored with ascorbic acid at -20℃

^**P＜0.01, ^***P＜0.001

Time of sonication (min)

A 0 1 5 15

Remaining percentage (%) of genipin

0 20 40 60 80 100

Fig. 7 Effect of thaw and sonication on genipin stability in serum (n=3).

(A) processed immediately without freeze and thaw

***

Time (min)

15 30 60 120 240

Remaining percentage (%) of genipin

0 20 40 60 80 100 120

Fig. 8 Effect of incubation time with sulfatase at 37℃ on genipin emergence in serum (n=3).

Minutes

Fig. 9 Chromatograms of genipin (GN) and methyl paraben (MP, internal standard) in

(A) rat feces upon spiking with geniposide without incubation

(B) rat feces spiked with geniposide and incubated at 37℃ for 1 min (C) blank rat feces

MP GN

Time (min)

0 5 10 15 20

Peak area ratio of (genipin/internal standard) ^6.5

7.0 7.5 8.0 8.5

Fig. 10 Mean (

±

S.D.) ratio-time profile of genipin after incubation of geniposide with rat feces (n=3).

Minutes

Fig. 11 Chromatograms of genipin (GN) and methylparaben (MP, internal standard) in

(A) plasma sample (iv) before hydrolysis (B) plasma sample hydrolyzed with sulfatase (C) blank rat plasma

(D) standard solution in MeOH

GN MP

Time (min)

0 100 2 0 0 300 4 0 0

Concentration of genipin (nmol/mL)

Fig. 12 Individual plasma concentration-time profiles of genipin (^●) and genipin sulfate (^○) in nine rats after intravenous bolus of genipin (50 mg/kg).

Time (min)

0 100 200 300 400

Concentration of genipin (nmol/mL)

Concentration of genipin (nmol/mL) ^0.1

1 mg/kg) to nine rats (left) and the semi-log diagram (right).

Time (min)

0 500 1000 1500 2000 2500 3000

Concentration of genipin (nmol/mL)

0 500 1000 1500 2000 2500 3000

Concentration of genipin (nmol/mL)

0.1 1

10 genipin 100 mg/kg

genipin 200 mg/kg

Fig. 14 Mean (

±

S.E.) plasma concentration-time profiles of genipin sulfate after oral administration of genipin (100, 200 mg/kg) to four rats (left) and the semi-log diagram (right).

Minutes

Fig. 15 Chromatograms of geniposide (GP) and caffeic acid (CA, internal standard) in

(A) GF decoction

(B) GF decoction without IS (C) standard solution in MeOH

GP GP GP

Time (min)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

Fig. 16 Individual plasma concentration-time profiles of genipin sulfate in six rats after oral administrations of GF decoctions (10, 20 g/kg).

Time (min)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0 2 4 6 8

GF 10 g/kg GF 20 g/kg

Time (min)

0 2000 4000 6000 8000 10000

Concentration of genipin (nmol/mL)

0.1 1 10

GF 10 g/kg GF 20 g/kg

Fig. 17 Mean (

±

S.E.) plasma concentration-time profile of genipin sulfate after oral administrations of GF decoctions (10, 20 g/kg)

Time (min)

0 1000 2000 3000 4000

Concentration of Methotrexate (µM)

0.00

MTX with GF decoction (4 g/kg) MTX with GF decoction (2 g/kg) MTX with GF decoction (1 g/kg)

Time (min)

0 1000 2000 3000 4000

Concentration of Methotrexate (µM)

0.001 0.01 0.1 1

MTX alone

MTX with GF decoction (4 g/kg) MTX with GF decoction (2 g/kg) MTX with GF decoction (1 g/kg)

Fig. 18 Mean (

±

S.E.) serum concentration-time profile of MTX after oral MTX alone (^●), and coadministration with 4 (^○), 2 (▼) and 1 g/kg (▽) of GF decoction (upper) and the semi-log diagram (lower).

M i n u t e s

Fig. 19 Chromatograms of PHT and 5,7-dimethoxycoumarin (5,7-DMC, internal standard)

(A) serum sample

(B) internal standard in blank rat serum (C) standard solution in MeOH

PHT

5,7-DMC

5,7-DMC PHT

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

0 PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

0 PHT with GF decoction

Fig. 20 Individual serum concentration-time profiles of PHT in six rats after oral PHT alone (^●), and coadministration with 2 g/kg (^○) of GF decoction.

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

0 2 4 6 8 10 12 14 16

PHT alone

PHT with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of PHT (µg/mL)

1 10

PHT alone

PHT with GF decoction

Fig. 21 Mean (

±

S.E.) serum concentration-time profile of PHT after oral PHT alone (^●), and coadministratio n with 2 g/kg (^○) of GF

M i n u t e s

Fig. 22 Chromatograms of CBZ and 5,7-dimethoxycoumarin (5,7-DMC, internal standard)

(A) serum sample

(B) internal standard in blank rat serum (C) standard solutio n in MeOH

5,7-DMC

5,7-DMC CBZ

CBZ

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 CBZ with GF decoction

Fig. 23 Individual serum concentration-time profiles of CBZ in six rats after oral CBZ alone (^●), and coadministration with 4 g/kg (^○) of GF decoction.

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

0 10 20

30 CBZ alone

CBZ with GF decoction

Time (min)

0 1000 2000 3000 4000

Concentration of CBZ (µg/mL)

1 10

CBZ alone

CBZ with GF decoction

Fig. 24 Mean (

±

S.E.) serum concentration-time profile of CBZ after oral CBZ alone (^●), and coadministration with 4 g/kg (^○) of GF decoction (upper) and the semi-log diagram (lower).

Table 1 The pH change of genipin solution (in water) titrated with 0.1N NaOH.

VNaOH (mL) 0.00 0.10 0.20 0.30 0.40 0.50 0.60

pH 5.63 5.85 6.02 6.16 6.30 6.47 6.68

V_NaOH(mL) 0.65 0.70 0.75 0.80 0.85 0.90 0.95

pH 6.77 6.91 7.06 7.48 8.35 8.79 9.03

V_NaOH(mL) 1.00 1.10 1.20 1.30 1.40

pH 9.17 9.40 9.57 9.69 9.80

Table 2 Intra-day and inter-day analytical precision and accuracy of genipin in plasma (n=3).

Precision Accuracy

Intra-day Inter-day Intra-day Inter-day

Conc.

(µg/mL) Mean ± S.D. (C.V.%) Mean ± S.D. (C.V.%) Relative error (%) Relative error (%)

10.0 10.0 ± 0.5 (5.2) 10.1 ± 0.1 (1.3) -0.1 0.5 5.0 5.0 ± 0.0 (0.3) 4.9 ± 0.0 (0.6) 0.0 -2.2

2.5 2.6 ± 0.0 (0.2) 2.5 ± 0.0 (0.0) 2.2 1.1

1.3 1.2 ± 0.0 (7.2) 1.2 ± 0.1 (6.4) -4.1 -3.4

0.6 0.6 ± 0.0 (1.9) 0.6 ± 0.0 (1.6) 2.6 3.9

0.3 0.3 ± 0.1 (1.3) 0.3 ± 0.0 (1.0) 1.1 6.6

0.2 0.2 ± 0.0 (1.9) 0.2 ± 0.0 (0.9) -1.5 12.5

0.1 0.1 ± 0.0 (1.1) 0.1 ± 0.0 (0.9) -16.3 11.5

Table 3 Recovery (%) of genipin from plasma (n=3).

Conc.

(

µ

g/mL) 1 2 3 Mean ± S.D.

5.0 100.1 99.6 100.3 100.0 ± 0.4 1.3 91.8 92.1 103.9 95.9 ± 6.9 0.3 100.3 100.4 102.5 101.1 ± 1.3

Table 4 Plasma concentrations (nmol/mL) of genipin in nine rats after intravenous bolus of genipin (50 mg/kg).

Time (min) 1 2 3 4 5 6 7 8 9 mean ± S.E.

5 32.6 45.7 51.9 70.7 48.9 47.7 60.4 40.3 68.2 51.8 ± 4.2 15 9.7 12.7 13.6 13.6 8.5 11.5 12.9 14.8 18.3 12.8 ± 0.9 30 3.7 4.7 4.0 1.1 2.4 1.8 1.3 2.4 3.5 2.8 ± 0.4 60 1.1 0.7 0.3 0.1 0.5 0.2 0.2 0.1 0.2 0.4 ± 0.1

Table 5 Plasma concentrations (nmol/mL) of genipin sulfate in nine rats after intravenous bolus of genipin (50 mg/kg).

Time (min) 1 2 3 4 5 6 7 8 9 mean ± S.E.

Table 6 Individual pharmacokinetic parameters of genipin in plasma after intravenous bolus of genipin (50 mg/kg) to nine rats.

Parameters 1 2 3 4 5 6 7 8 9 Mean ± S.E.

Table 7 Individual pharmacokinetic parameters of genipin sulfate in plasma after intravenous bolus of genipin (50 mg/kg) to nine rats.

Parameters 1 2 3 4 5 6 7 8 9 Mean ± S.E.

Cmax (nmol/mL) 17.5 26.1 38.2 12.9 7.5 12.7 12.2 6.6 6.9 15.6 ± 3.5 AUC0-t

(nmol•min/mL) 375.1 347.2 584.3 274.1 212.7 346.3 315.9 228.9 400.2 342.7 ± 36.9 Cl (mL/min) 179.5 190.6 108.1 266.5 362.1 234.7 245.6 311.1 166.3 229.4 ± 26.0 V (mL) 12293.3 24923.0 8924.5 31506.9 66598.5 45809.8 10198.2 72121.8 20108.2 32498.2 ±7978.4 MRT0-t (min) 44.1 31.7 23.7 23.4 38.2 26.4 27.0 69.3 86.7 41.2 ± 7.5

Cmax：藥物在血中之最高濃度；血峰濃度

Table 8 Pharmacokinetic parameters of genipin parent from and genipin sulfate in

在文檔中梔子中Geniposide 於大鼠之代謝動力學及其與西藥之交互作用; Metabolic Pharmacokinetics of Geniposide in Gardenia Fruits and Interactions with Western Medicines in Rats (頁 102-147)

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