手性药物代谢的立体选择性 - njdmpk.com · 药物代谢酶 1.2万名“海豹 ......
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临床使用药物中手性药物比例
Single
Enantiomer / One
Chiral Center
Racemates
Non-Chiral
Single
Enantiomer /
Multiple Chiral
Centers
35%
20%35%
10%
全球500种畅销药物中手性药物占59%
59%
手性药物 比例高 疗效好
一、背 景
震惊世界的反应停事件
N
N OO
O
O
H
N
CO2H CO2H
O
O
(-)-S-thalidomide(-)-S-phthalimidoglutamic acid
(RS)-反应停S-反应停 S-苯二酰亚氨基谷氨酸
毒性对映体代谢物毒性对映体杂质
药物代谢酶
1.2万名“海豹形”婴儿
� 药物手性杂质和代谢物
� 即使仅微量存在,也可能产
生各种毒性
� 直接影响到新药研发成功率
及药品的安全性和有效性
� 关系到用药者的健康与生命
安危
药 物 对映体相对活性
特步他林 - > + (3000:1)
吲哚洛尔 - > + (200:1)
普萘洛尔 S > R (100:1)
维拉帕米 - > + (18:1)
华法令 S > R (8:1)
氯胺酮 S > R (4:1)
美沙酮 - > + (3:1)
手性药物的“错误”对映体,不但无效甚至有严重的毒副作用
药名 S- R-
四咪唑 驱虫免疫增强 呕吐
青霉胺 抗关节炎 潜在致癌性
乙胺丁醇 抗结核 失明
酞胺哌啶酮 镇静 致畸
各国手性药物研究技术法规状况
� 美国FDA: Development of New Stereoisomeric Drugs,
1992
� 欧洲EMEA: Investigation of chiral active substances,
1994
� 加拿大Canada Minister of Health, Guidance For
Industry: Stereochemical Issues in Chiral Drug
Development, 2000
� 中国,手性药物质量分析指导原则,2012年
8 July 11, 2016
肝脏微粒体酶 (氧化,结合)
肝外微粒体酶 (氧化,结合)
肝脏非微粒体酶 (乙酰化,硫酸化, 醇醛脱氢酶,GST, 水解,氧化/还原) 生物体的手性环境
Absorption
Distribution
Metabolism
Excretion
手性药物代谢
手性高效液相色谱法
手性气相色谱法
手性色谱学 手性毛细管电泳法
手性超临界色谱法
手性薄层色谱法
间接法: (Chiral Derivatization Reagent, CDR)
色谱法
直接法: (Chiral Mobile Phase Additives, CMPA)、
(Chiral Stationary Phase, CSP)
对映体选择性测定方法
Yu L, Wang S, Jiang H, Zhou H, Zeng S. Simultaneous determination of fluoxetine and norfluoxetine enantiomers using isotope discrimination mass
spectroscopy solution method and its application in the CYP2C9-mediated stereoselective interactions. J Chromatogr A. 2012 May 4;1236:97-104.
HO
HO
CO2H
NH2
N
N N
NN
CO2H
O CO2H
Me
NH2
H2N
S
N
NPh
NH2
O
CO2H
MeO
H H H HO
OH
NBu
OHH
t
L-dopa methotrexate
cephalexin terbutaline
Drugs With Stereoselective Absorption
Cl
N
N
O COOH
Cl
N
N
O COOH
1. ABSORPTION
cetirizine
西替利嗪对映体双向转运试验
A B
Transport rate of cetirizine enantiomers across Caco-2 cell monlayers in the
absorptive (A) and secretary (B) direction. The insets represented the
Eadie-Hofstee transformation of the data for the secretary direction
0
200
400
600
800
0 20 40 60 80C(µmol/L)
Tra
nsp
ort
Rat
e(×
10
-9m
olc
m-2
s-1)
R-cetirizine
S-cetirizine
0
400
800
8 10 12 14V/C
V
0
100
200
300
400
500
0 20 40 60 80C(µmol/L)
Tra
nsp
ort
Rat
e(×
10
-9m
olc
m-2
s-1)
R-cetirizine
S-cetirizine
2. DISTRIBUTION
Drug Free Fraction Ratio (+/-)
+ -
disopyramide 0.27 0.39 0.7
ibuprofen 0.006 0.0039 1.5
mexilitine 0.283 0.198 1.4
methadone 0.092 0.124 0.7
propoxyphene 0.018 0.018 1.0
propranolol 0.203 0.176 1.2
tocainide 0.17 0.14 1.2
verapamil 0.064 0.11 0.6
warfarin 0.012 0.009 1.3
O
OH
O
Ph
Et
O
OH
O
Ph
Me
O
Cl N
NO
Cl
OH
H
NN
Ph
Ph
O
O
SPh
O
Cl N
NO
Ph
H
H
OCOCH2CH2CO2H
Cl N
N
Ph
OH
MeO
MeO
N
N
Me
MeOOMe
Et
Cl N
MeO
Ph
H
N
CO2H
NH2
H
NCO2H
Cl
H
Me O
Ph CO2H
Me
PhCO2H
Me
phenprocoumon (-)-S-warfarinS-lorazepam
sulfinpyrazone S-oxazepam succinate des-methyl diazepam
S-tofisopam 4,5-dihyddrodiazepam L-tryptophan
(+)-S-carprofen ketoprofen fenoprofen
Chiral Drugs
Stereoselectively
Binding with
HSA
0
10
20
30
40
50
1 2
Bound f
ract
ion(%
)
The binding of S-ketoprofen (1) and R-ketoprofen (2)
(1.25 µmol·mL-1) to HSA DOM III (5.2 µmol·mL-1)
(n = 3)
S-ketoprofen
R-ketoprofen
*
N
N
O O
H O
H
H
( 1)N
NO O
PhPhH
H
(2) N
NOO
O H
H
H
(-)-S - d iphenylhydanto in m etabolism (+)-R -
S + · ·O
Et
M e
FID
S + · ·· ·
E t
M e
S + · ·· ·
E t
M e
a
(+)-R -4-to ly l e thyl su lph ide m etabolism (-)-S -
b
N
O O
M e
H
Et
N
O O
M e
O H
H
Et
N
O O
M e
H
E t
N
O O
M e
H
E t
S -
R -
c m epheneto in m etabolism
most of species
CYP
(1:4 R:S)
S-enantiomer
R-enantiomer
dog
FAD
(20:1 R:S)
N
HMe
NH
HMe
NH2
HMe
NHNH
Pt
O
Me
NH2
Me
a (S)-(+)-deprenyl (S)-(+)-methamphetamine (S)-(+)-amphetamine
b (R)-(-)-prilocaine 0-toluidine
对映体生物转化增加毒性
Toxicity!
S
R
R'
S
R
R'
O[O]
C
C
H
HR
R'
C
OH
HR
R'[O]
A
C O
R'
R[H]
C
OH
HR'
R
D
C C
R H
R"R'
[H]C C
HR
H R"H R'E
N
R
R'
R"
[O]N
R
R'
R"
OB生物体内
产生手性中心的代谢反应
A.亚甲基氧化
B.叔胺氧化
C.硫化物氧化
D.酮基还原
E.双键还原
H3C
CH3 COOH
H
H3C
S-Ibuprofen
H3C
CH3 COOH
H
H3C
R-Ibuprofen
Inhibits platelet thromboxane production twice as effectively as does the racemate
Racemizes to the S-form and does not cause G.I. lesions
布洛芬 服用的S/R比例 排泄的S/R比例
(S)-(+) 95:5 95:5
(R)-(-) 6:94 80:20
外消旋体 50:50 70:30
手性转化增加毒性
4. EXCRETION
Drug Clearance Form (+) (-) Ratio
chloroquine CLR,u racemate 824 519 1.6
Disopyramide CLR,u isomer 338 182 1.8
mexilitine CLR racemate 0.5 0.5 1.0
pindolol CLR,u racemate 453 534 1.2
terbutaline CLR isomer 2.7 1.5 1.8
tocainide CLR isomer 55 55 1.0
Drug Clearance Form (+) (-) Ratio
propranolol CLiv racemate 1.21 1.03 1.17
CLu,iv racemate ~6.0 ~5.9 1.02
CLo racemate 2.78 1.96 1.42
CLu,o racemate 13.7 11.1 1.23
verapamil CLiv isomer 0.80 1.40 1.75
CLu,iv isomer 5.96 5.85 1.02
CLo racemate 1.72 7.46 4.33
warfarin CLo racemate 0.234 0.333 1.42
CLu,o racemate ~21.9 ~37.0 1.69
� Peak 1: R-(+)-PPL glucuronide
� Peak 2: S-(-)- PPL glucuronide
� Peak 3: RS-PPL
Blank without
UDPGAUGT2B7 UGT1A9
Stereoselectivity of Propranolol side chain(-OH) glucurnidation
0
50
100
150
200
250
5 10 20 40 60 80
Time(min)
Propranolol glucuronide
produced(pmol)
R-PL-Glu(UGT1A9)
S-PL-Glu(UGT1A9)
R-PL-Glu(UGT2B7)
S-PL-Glu(UGT2B7)
O
OH
N CH3
CH3
HO
UDPGT O
O
N CH3
CH3
OUDP OH
OH
COOH
HO UDP
O
OH
OH
COOH
HO
H
Chirality. 2010;22:456-61.
The cumulative excretion percentage of S-(-)- and R-(+)-
propranolol glucuronide in Chinese Han subjects urines
after 20mg oral administration of RS-(±)-propranolol
tablet (x±s%, n=16)
0
5
10
15
20
0 10 20 30
Time(h)
The cumulative
excretion%
S-glucuronide
R-glucuronide
UGT1A9 play a key role for stereoselectivity of Propranolol glucurnidation
28/39
6对洛芬类NSAIDs对映体HN
ClHO
O
CH3H HN
Cl HO
O
HH3C
O
HO
O
CH3H
O
HO
O
HH3C
HO
O
CH3
H
F
HO
O
HCH3
F
H3C H
OH
O
H CH3
OH
O
OH
O
O
CH3H
OH
O
O
HH3C
S
O
OH
O
CH3H
S
O
OH
O
HH3C
CPF
FePF
FPF
IPF
KPF
ZPF
30/39
UGT2B7*1
UGT2B7*71S(C3Y)
UGT2B7*2(H268Y)
UGT2B7*5(D398N)
UGT1A9*1
UGT1A9*2(C3Y)
UGT1A9*3(M33T)
UGT1A9*5(D256N)
Wang H, Yuan L, Zeng S. Characterizing the effect of UGT 2B7 and UGT1A9 genetic polymorphisms on enantioselective glucuronidation of flurbiprofen.
Biochem Pharmacol. 2011;82:1757-63.
UGT1A9和2B7对氟吡洛芬的代谢
� UGT1A9活性顺序为*2>*1,*5,突变体立体选择性增加(S/R=0.6-
0.3) , UGT2B7活性顺序为*1>*71S>*2,立体选择性不变
(S/R=2.2-1.8),个体化用药依据
� 二者对FPF代谢的立体选择性相反
氟吡洛芬