药代动力学在新药研发中的作用
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Final concentrations of test article = 0.05-0.30 mg/mL
in situ rat intestinal perfusion (single pass)
Perfusion Procedures:
• rat is put on a heating pad to maintain body temperature • jejunum is exposed via a middle line incision • sutures: 1st is made at 5 cm distal to the ligament of Treitz
Animal: Male Sprague-Dawley rats (250 - 350 g), fasted overnight.
Rat is anesthetized by urethane 1.5g/kg, im. before perfusion starts.
Perfusate: Phosphate buffer, pH = 6.5 10 mM glucose Phenol red (negative control) Acetaminophen (positive control)
Caco-2 Transport Pathways 人大肠癌细胞吸收模型
False Positive 假阳性 = 低
False Negative 假阴性 = 高
in situ rat intestinal perfusion (single pass) 离体大鼠十二指肠灌流模型(单循环)
METHOD
药物代谢及其动力学在新药研发中的应用
胡卓汉 博士 瑞德肝脏疾病研究(上海)有限公司
复旦大学药学院
2004年12月30日 中国.北京
药物研发的三大任务
• 药效
Efficacy/ Pharmacodynamics • 安全
Safety / Toxicology • 药物代谢动力学
Drug Metabolism/Pharmcokinetics
Drugs
caffeine ibuprofen desipramine acetaminophen propranolol hydrazine Ketoconazole terbutaline atenolol acetbutalol nadolol losartan mannitol inulin
AUC ( g/m l/m in) 4.823
Bioavailability % 2.523
R2
0.9292
M e tabolite
6.53 0.799 21.810 11.434 0.9964
M e tabolite BCH3840
60
ห้องสมุดไป่ตู้
120
180
Time (min)
Plasma concentrations of BCH-3840 and its metabolite (BCH-6440) in mice dosed 50 mg/kg orally
Glucose
• P-Glycoprotein mediated (P-糖蛋白调节)
底物
Vinblastine
抑制物 Verapamil
Glucose (蔗糖) vs Inulin (木香素) 主动吸收 vs 细胞间渗透
Flux
250 200 150 100 50
0 0
20 40 60 Time (min)
NDA 临床实验
非临床实验准则
Good Laboratory Practice (GLP)
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
二五原则
• 5 毫克 •5天
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
药物代谢动力学的任务
血浆浓度
100 10 1 0 .1
0 .0 1 0
(最大无毒性浓度)
80 100
Propranolol vs Mannitol 被动吸收 vs 细胞间渗透
Papp (nm/sec)
450
Mannitol
400
Propranolol
350
300
250
200
150
100
50
0
day1 1999-8- 1999-9- 1999-9- 1999-9- 1999- 1999- 1999- 1999- 1999- 1999- 2000-1- 2000-1- 2000-1-
80
BE study 2 specification study
60
40
20 Y = 4.2 + 1.00 X, R2 = .987
0
0
20
40
60
80
100
% Distributed
15
Caco-2 Transport Pathways 人大肠癌细胞模型
Transport Pathways 药物吸收机制
extrapolation
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits R&D
Optimized Lead
Go or no go decision
研究和发现
临床阶段
•长期毒性实验的动物选择
metabolism profiling in animals and humans
被动
细胞间 主动
P糖蛋白
Probes for Transport Pathways 肠道吸收标准对照药物
• Transcellular (被动吸收)
Propranolol, Testosterone
• Paracellular (细胞间渗透)
Mannitol, Inulin
• Carrier mediated (主动吸收)
Oral Absorption in Humans (%)
100 100 95 95 90 90 76 73 50 40 35 33 16
5
3-day Caco-2
Kp
227 201 261 218 265 155 120 56 30 29 22 42 40 12
Chong, Dando & Morrison; Pharm. Res. 1997
8/23/99 30
13
21
28 10-5 11-1 11-8 12-6 12-13 12-20 10
17
24
Test Day
由P-蛋白所调节的药物吸收 -使用P-糖蛋白抑制剂 Verapamil
Papp (nm/sec)
120
Vinblastine
100
Vinblastine/verapa
80
mil
in situ离体
permeability
in vivo体内
bioavailability
Poor oral bioavailability
Concentration [ng/ml]
1000 800 600 400 200 0 0
Par e nt
Tm ax (m in)
6.14
Cm ax ( g/m l) 0.083
60
40
20
0
day1 1999-8- 1999-9- 1999-9- 1999-9- 1999-
8/23/99 30
13
21
28 10-5
199911-1
199911-8
199912-6
199912-13
1999- 2000-1- 2000-1- 2000-1-
12-20 10
17
24
Test Day
• 对其它药物的影响? drug-drug interaction
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
临床前阶段
• 生物利用度
Calculations:
Permeability (Peff, cm/min) = (Q/2RLp) x (1- C’out / C’in )
C’out / C’in = (Cout / Cin) x [ phenol red ] in / [ phenol red ] out
In situ rat intestinal permeability (single pass)
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
临床实验准则
Good Clinical Practice (GCP)
NEW DRUG
2nd is made at about 20 cm distal to 1st one • the inlet of cannula - a syringe infusion pump • the outlet of cannula - a fraction collector • the perfusion segment is precleaned by passing 10 ml of blank perfusate buffer • perfusion time and rate = 0.1 ml/min for 120 min • outlet perfusion samples are collected every 10 min • plasma samples are collected at 30, 60, 90 and 120 min after perfusion
动物推测人 (species extrapolation)
Situation Analysis
后期研发阶段
早期研发阶段
排出太快/药效时间太短
口服吸收差/血浆浓度太低
分布
排泻
代谢问题
吸收问题
蛋白质相互作用
分布体积
肾脏排泄
肝脏代谢
肠道消化
溶解度
肠道吸收 膜通透性
in vitro体外
metabolism
pH=7.4
14
in vitro absorption/distribution model
In Vitro/ In Vivo Correlation Pooled Data from Four Biostudies
% Absorped
100
formulation finding study
BE study 1
In-house validation
Human Oral Bioavailability (%)
100
假阴性
80
60
40
20 0
0.000
假阳性
0.001 0.002 0.003 0.004
Permeability (cm/min)
bioavailability • 血浆浓度的线性和非线性
dose escalation & proportionality • 多次给药和体内积蓄
multiple doses & accumulation • 吸收和排泄模式
mass balance • 体内分布
distribution • 从动物代谢推算人体代谢
药物吸收模型
计算机 脂溶度 脂层转移 细胞层转移 十二指肠灌流
Permeability Evaluation – in vitro
absorption/distribution model 脂层转移模型
有机相 Organic phase
水相 Aqueous phase
pH=6.5
水相 Aqueous phase
临床前实验药物代谢动力学的生物模型
体外和离体模型 (in vitro / in situ models) •吸收模型 absorption/permeability •代谢模型 metabolism •体外推测和体内 (in vitro / in vivo correlation)
动物模型 (in vivo animal models)
(最小药效时间)
0 .5
1
1 .5
时间2.
2 .5
(最小有效浓度)
3
3 .5
4
药效
最佳 血浆浓度
毒理
药代
研究和发现阶段
• 能否被吸收? permeability
• 是否被代谢? metabolic stability
• 代谢产物? metabolite identification
• 代谢途径? pathway identification
in situ rat intestinal perfusion (single pass)
Perfusion Procedures:
• rat is put on a heating pad to maintain body temperature • jejunum is exposed via a middle line incision • sutures: 1st is made at 5 cm distal to the ligament of Treitz
Animal: Male Sprague-Dawley rats (250 - 350 g), fasted overnight.
Rat is anesthetized by urethane 1.5g/kg, im. before perfusion starts.
Perfusate: Phosphate buffer, pH = 6.5 10 mM glucose Phenol red (negative control) Acetaminophen (positive control)
Caco-2 Transport Pathways 人大肠癌细胞吸收模型
False Positive 假阳性 = 低
False Negative 假阴性 = 高
in situ rat intestinal perfusion (single pass) 离体大鼠十二指肠灌流模型(单循环)
METHOD
药物代谢及其动力学在新药研发中的应用
胡卓汉 博士 瑞德肝脏疾病研究(上海)有限公司
复旦大学药学院
2004年12月30日 中国.北京
药物研发的三大任务
• 药效
Efficacy/ Pharmacodynamics • 安全
Safety / Toxicology • 药物代谢动力学
Drug Metabolism/Pharmcokinetics
Drugs
caffeine ibuprofen desipramine acetaminophen propranolol hydrazine Ketoconazole terbutaline atenolol acetbutalol nadolol losartan mannitol inulin
AUC ( g/m l/m in) 4.823
Bioavailability % 2.523
R2
0.9292
M e tabolite
6.53 0.799 21.810 11.434 0.9964
M e tabolite BCH3840
60
ห้องสมุดไป่ตู้
120
180
Time (min)
Plasma concentrations of BCH-3840 and its metabolite (BCH-6440) in mice dosed 50 mg/kg orally
Glucose
• P-Glycoprotein mediated (P-糖蛋白调节)
底物
Vinblastine
抑制物 Verapamil
Glucose (蔗糖) vs Inulin (木香素) 主动吸收 vs 细胞间渗透
Flux
250 200 150 100 50
0 0
20 40 60 Time (min)
NDA 临床实验
非临床实验准则
Good Laboratory Practice (GLP)
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
二五原则
• 5 毫克 •5天
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
药物代谢动力学的任务
血浆浓度
100 10 1 0 .1
0 .0 1 0
(最大无毒性浓度)
80 100
Propranolol vs Mannitol 被动吸收 vs 细胞间渗透
Papp (nm/sec)
450
Mannitol
400
Propranolol
350
300
250
200
150
100
50
0
day1 1999-8- 1999-9- 1999-9- 1999-9- 1999- 1999- 1999- 1999- 1999- 1999- 2000-1- 2000-1- 2000-1-
80
BE study 2 specification study
60
40
20 Y = 4.2 + 1.00 X, R2 = .987
0
0
20
40
60
80
100
% Distributed
15
Caco-2 Transport Pathways 人大肠癌细胞模型
Transport Pathways 药物吸收机制
extrapolation
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits R&D
Optimized Lead
Go or no go decision
研究和发现
临床阶段
•长期毒性实验的动物选择
metabolism profiling in animals and humans
被动
细胞间 主动
P糖蛋白
Probes for Transport Pathways 肠道吸收标准对照药物
• Transcellular (被动吸收)
Propranolol, Testosterone
• Paracellular (细胞间渗透)
Mannitol, Inulin
• Carrier mediated (主动吸收)
Oral Absorption in Humans (%)
100 100 95 95 90 90 76 73 50 40 35 33 16
5
3-day Caco-2
Kp
227 201 261 218 265 155 120 56 30 29 22 42 40 12
Chong, Dando & Morrison; Pharm. Res. 1997
8/23/99 30
13
21
28 10-5 11-1 11-8 12-6 12-13 12-20 10
17
24
Test Day
由P-蛋白所调节的药物吸收 -使用P-糖蛋白抑制剂 Verapamil
Papp (nm/sec)
120
Vinblastine
100
Vinblastine/verapa
80
mil
in situ离体
permeability
in vivo体内
bioavailability
Poor oral bioavailability
Concentration [ng/ml]
1000 800 600 400 200 0 0
Par e nt
Tm ax (m in)
6.14
Cm ax ( g/m l) 0.083
60
40
20
0
day1 1999-8- 1999-9- 1999-9- 1999-9- 1999-
8/23/99 30
13
21
28 10-5
199911-1
199911-8
199912-6
199912-13
1999- 2000-1- 2000-1- 2000-1-
12-20 10
17
24
Test Day
• 对其它药物的影响? drug-drug interaction
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
临床前阶段
• 生物利用度
Calculations:
Permeability (Peff, cm/min) = (Q/2RLp) x (1- C’out / C’in )
C’out / C’in = (Cout / Cin) x [ phenol red ] in / [ phenol red ] out
In situ rat intestinal permeability (single pass)
NEW DRUG
NDA 临床实验
Compound for Development (CD)
IND 临床前实验
Efficacy Hits
R&D
Optimized Lead
Go or no go decision
研究和发现
临床实验准则
Good Clinical Practice (GCP)
NEW DRUG
2nd is made at about 20 cm distal to 1st one • the inlet of cannula - a syringe infusion pump • the outlet of cannula - a fraction collector • the perfusion segment is precleaned by passing 10 ml of blank perfusate buffer • perfusion time and rate = 0.1 ml/min for 120 min • outlet perfusion samples are collected every 10 min • plasma samples are collected at 30, 60, 90 and 120 min after perfusion
动物推测人 (species extrapolation)
Situation Analysis
后期研发阶段
早期研发阶段
排出太快/药效时间太短
口服吸收差/血浆浓度太低
分布
排泻
代谢问题
吸收问题
蛋白质相互作用
分布体积
肾脏排泄
肝脏代谢
肠道消化
溶解度
肠道吸收 膜通透性
in vitro体外
metabolism
pH=7.4
14
in vitro absorption/distribution model
In Vitro/ In Vivo Correlation Pooled Data from Four Biostudies
% Absorped
100
formulation finding study
BE study 1
In-house validation
Human Oral Bioavailability (%)
100
假阴性
80
60
40
20 0
0.000
假阳性
0.001 0.002 0.003 0.004
Permeability (cm/min)
bioavailability • 血浆浓度的线性和非线性
dose escalation & proportionality • 多次给药和体内积蓄
multiple doses & accumulation • 吸收和排泄模式
mass balance • 体内分布
distribution • 从动物代谢推算人体代谢
药物吸收模型
计算机 脂溶度 脂层转移 细胞层转移 十二指肠灌流
Permeability Evaluation – in vitro
absorption/distribution model 脂层转移模型
有机相 Organic phase
水相 Aqueous phase
pH=6.5
水相 Aqueous phase
临床前实验药物代谢动力学的生物模型
体外和离体模型 (in vitro / in situ models) •吸收模型 absorption/permeability •代谢模型 metabolism •体外推测和体内 (in vitro / in vivo correlation)
动物模型 (in vivo animal models)
(最小药效时间)
0 .5
1
1 .5
时间2.
2 .5
(最小有效浓度)
3
3 .5
4
药效
最佳 血浆浓度
毒理
药代
研究和发现阶段
• 能否被吸收? permeability
• 是否被代谢? metabolic stability
• 代谢产物? metabolite identification
• 代谢途径? pathway identification