1. 연구분야 보유기술 의약후보물질 최적화를 위한 독성평가 화학물질의 체내 대사체 구조 규명 (1상 및 2상 대사체)
화학물질의 체내 대사경로 연구
장내 미생물에 의한 화학물질 대사 연구
약물 상호작용 연구
보유기술
간독성 및 면역독성 평가기술 (in vitro 및 in vivo)
대사체 구조 결정 (LC-MS)
약물 상호작용 평가 기술
간세포 분리 및 일차배양 기술
CYP 효소활성 측정기술

2. 연구분야 소개
Metabolism
The elimination of xenobiotics depends on their conversion to water-soluble metabolites by a process known as biotransformation, which is catalyzed by enzymes in the liver and other tissues.

3. Drug metabolite with modified activity Inactive drug metabolite
Phase Ⅰ and Phase Ⅱ Reactions
ABSORPTION
METABOLISM
ELIMINATION
Phase Ⅰ
Phase Ⅱ
Conjugate
Drug metabolite with modified activity
Urine
Feces
Inactive drug metabolite
  Lipophilic
Hydrophilic

4. UV Chromatograms of Rutaecarpine and its Metabolites
without NGS
Rutaecarpine
with NGS
Rutaecarpine
Metabolites

5. MS2 Spectra of Parent: Rutaecarpine
m/z 169 EE+
Rutaecarpine [M+H]+ 288
m/z 120 EE+
m/z 142 EE+
m/z 145 EE+
m/z 185 OE+
145.0
142.0
185.0
[M+H]+
288
120.0
168.9

6. Proposed Metabolic Pathway of Rutaecarpine in Human Liver Microsomes
β-Glucuronidation and Sulfation
β-Glucuronidation and Sulfation M1
Only in vitro
CYP3A4 M2 M5
CYP3A4
CYP3A4/1A2/2C9
CYP3A4/1A2/2C9
CYP1A2/2C9/3A4
Rutaecarpine M3 M4
β-Glucuronidation and Sulfation

7. Acute effects of 1-BP on serum activity of ALT and histopathology of liver tissue
Vehicle
200 mg/kg
500 mg/kg
1000 mg/kg
6 hr
24 hr
48 hr
12hr
X 100 magnification

8. T-dependent antibody response to SRBC
SRBCs (5×108)
After 30 min
4 days later
1-BP treated
BALB/c mice, po
Sensitization, ip
Single
splenocyte
Spleen
37oC
3 h later
0 h
Splenocytes,
SRBC, Complement
(guinea pig) and 0.5%
agar containing
0.05% DEAE-dextran
Stereomaster plaque viewer, ×20
AFCs
Splenocytes counting
(Coulter counter)
Cover glass
(24×40 mm)
Results:
AFCs/106 spleen cells
or AFCs/spleen (×103)
Petri dish (100×15 mm)

9. CID Spectrum of Glutathione+
+H+ a b c
[M+H-129]+
100
150
200
250
300
350
400
m/z 10 20 30 40 50 60 70 80 90
Relative Abundance
179.0
162.0
233.0
308.0
290.1
Glutathione [M+H]+ 308 a b c
[M+H-146]+
[M+H-75]+
[M+H]+

10. Fragmentation Mechanism and Proposed Structures of GSH+
+H+ b c c
Loss of neutral glycine
[M+H]
[M+H-75] b a
Loss of amino glutamate
Loss of glutamate +
+H+
[M+H-146]
[M+H-129]

11. Metabolism of 1-BP Excretion in urine GST CYP GST CYP GST CYP GST GST
N-acetyl-S-propyl cysteine
S-propyl glutathione, m/z 350
Dehydrohalogenation (Enzyme-mediated)
1,2-Propane-diol (Ether extract) NADPH- dependence
Propene (Volatile metabolite) NADPH- dependence
CYP
GST
1-Bromopropane
CYP
GST
α-hydroxylation
N-acetyl-S-(2-hydroxypropyl)cysteine
S-(2’-Hydroxyl-1’-propyl) glutathione
CYP
3-Bromo-2-propanol
ALDH
Propionic acid (Ether extract) NADPH- dependence
Propionaldehyde
C3 oxidation
GST
Excretion in urine
3-Bromo-1-propanol
N-acetyl-S-(3-hydroxypropyl)cysteine
GST
3-Bromopropionic acid
Barnsley et al., 1966, Jones and Walsh, Tachizawa et al., 1982
N-acetyl-S-(2-carboxyethyl) cysteine

12. Effects of 1-BP on content of GSH and formation of S-propyl GSH in liver and spleen: Dose-response

13. ? Overall ranking of toxicity:
stabilizer the wool
chemical syntheses
cleaning agent
Overall ranking of toxicity:
1,2-DBP, 1,3-DBP > 1-BP >> 2-BP
extraction solvents
Increased the serum ALT activity and tissue damage
Liver
spray form
chemical intermediates
Exposure
Depletion of GSH
Increased oxidative stress
Suppressed the antibody response to SRBCs
Suppressed the splenic intracellular IL-2 production
Spleen ?
Dependent mechanism
Glutathione
S-propyl GSH (1-BP)
S-Isopropyl GSH (2-BP)
S-2-hydroxypropyl(oxopropyl) GSH
& their mercapturic acid (1,2-DBP)
S-2-79/81bromopropenyl GSH (2,3-DBPE)
S-3-79/81bromopropyl GSH (1,3-DBP)