优秀学术报告PPT示例(深底色)(杉山雄一)

S
S
HO
O
S
S
N H O
N COOH
N H O
N COOH
temocapril
temocaprilat
Chemical structures of temocapril and temocaprilat
100 Plasma concentration (ng/ml) 80 60 40
temocaprilat
out
in NH 2 COOH
Oat 1 (551 a.a. M.W. 60kDa)
12 TMDs Tissue distribution: kidney (basolateral membrane) Substrates: p-aminohippurate, dicarboxylates, PGE2, urate, NSAIDs, -lactam antibiotic
Variability
Excretion
(Liver Kidney)
Protein Gene
Metabolism
(Phase II) Analysis of Relationship between Each Process
Species Difference
Genetic Polymorphism
MRP3 OATPs Oatp2/ OATP-A Mrp1
Absorption
Oatp2/OATP-A Oatp2/OATP-A Oat3
Blood 【Kidney】
OAT1 OAT3 OCT2
Blood
Bcrp
P-gp/ MDR1
Mrp1
NTCP
Oatp1,2,4/ OATP-C,B,8
OCT1
OAT2 MRP3
CLcr<40 40<CLcr<82 82<CLcr
50 40 30 20 10 0
enalaprilat
20
0 0 12 24 36 Time (hr) 48
0
12
24 Time (hr)
36
48
Plasma concentration of ACE inhibitors in patients with various renal functions
？ Байду номын сангаасbsorpt ion
(OATP-B ?)
Small Intestine
Coordination of
OATP2/OATP-C and MRP2 / cMOAT
（（（（9 分
Liver specific expression of OATP2 in Northern blotting
E 217β G、 E1S TCA、 pravastatin
Glucuronide Conjugate
Bile acids
Others
LTC4 DNP-SG glutathione bimane glutathione disulfide (GSSG) monochloro-monoglutathionyl melphalan 17-estradiol 17-( -D-glucuronide) -naphthyl glucuronide bilirubin glucuronide E3040 glucuronide liquiritigenine (flavonoid) glucuronide glycyllhizin （ flavonoid-glucuronide） grepafloxacin (new quinolone) glucuronide SN-38 (camptothecin analogue)s glucuronide glucuronosyl etoposide cholate-3-O-glucuronide lithocholate-3-O-glucuronide tauro/glycolithocholate 3-sulfate taurochenodeoxycholate 3-sulfate 6a -Glucuronosylhyodeoxycholate 3a -sulfatolithocholyltaurine LTE4 LTD4 LTE4NAc dibromosulfophthalein cefodizime (-lactam antibiotic) grepafloxacin (new quinolone antibiotic) methotrexate CPT-11 acid form (camptothecin analogue) SN-38 acid form (camptothecin analogue) pravastatin (HMA CoA reductase Inhibitor) temocaprilat (ACE inhibitor) BQ-123 (cyclic peptide; endothelin antagonist)
Met abolism
ATP ADP
Hepat ic upt ake
Biliary excret ion cMOAT ((MRP2)
(OATP2) oat p
Binding t o HMG-CoA reduct ase
Liver Enterohepatic circulation
pravastatin
NH 2 COOH out in
Mrp1 (1500 a.a. M.W. 180~190 kDa)
17 TMDs Tissue distribution: lung, muscle, testis, brain Substrates:glutathione conjugates, glucuronides, vincristine (+GSH)
40 Uptake (µl/mg protein)
Normal
(A) Uptake (µl/mg protein)
40
(B)
EHBR EHBR
30
30 +ATP 20
20 +ATP 10
10 -ATP 0 0 10 20 Time (min) 30
-ATP
0 0 10 20 Time (min) 30
Approach for Elucidation of Detoxification Mechanism
Procedure for Genomic Drug Discovery
Pharmacokinetics CYP enzymes Transporters
Genomic Analysis
Identification of the disease associated gene
Hepatic Drug Metabolic Enzymes/Transporters ： Effect on hepatic clearance and bioavailability
Yuichi Sugiyama
04.12.30 Dept of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences The University of Tokyo
Biopharmaceutics
Identification of lead compounds
Lead Optimization
Combinatrial Chemistry（CC） Screening of the physicochemical property High Throughput Screening（HTS） In silico screening (Virtual screening) Utilization of Toxicogenomics Analysis of the genetic polymorphism （SNPs etc.） Functional Analysis of the mutated proteins
SPGP/ BSEP
MRP2
Xenobiotics detoxification system in Human Liv
Analysis of Each Process
Metabolism
(Phase I) Whole body Organ Cell Elucidation昉 rediction Detoxification Mechanism
Time profiles of [14C] temocaprilat into CMVs
ACE Inhibitors
temocaprilat
OATP2 MRP2
captopril enalaprilat etc.
Urine
Bile
Discrimination based on the difference in pharmacokinetic profile
Brain endothelial cells
P-gp/ MRP2,4 MDR1
OCTN1,2
PepT2
OAT4
【Blood brain barrier】
P-gp/ MRP2 MDR1 BCRP BSEP
Urinary excretion
Importance of vectorial transport
【Liver】
Biliary Excretion
Major drug transporters expressed in the liver,kidney, small intestine and brain
Importance of vectorial transport in the liver
Identification of the target gene
Bioinformatics Functional genetic analysis Utilization of transgenetic animals Analysis of the genetic polymorphism （SNPs etc.） Structural Biology
Bile acids Na +
OCT1 NTCP
OATP-2/ OATP-C LST-1
OATP-8
OATPA, D, E?
OATP-B
OAT2 MRP3
ATP
ADP
Phase I 代謝 -OH Phase II 代謝 -OX
ATP ADP ATP ADP
ATP
MRP6
ATP ADP ADP
MDR1
ACE inhibitor： Temocaprilat
HMG CoA-reductase inhibitor Pravastatin、pitavastatin, rosuvast Angiotensin receptor antagonist Valsartan, Termisartan
H3C
O
O
ATP
ATP
MDR1
human hepatocytes hepatocyte human
Li ve
r
Substrates for rat MRP2 and human MRP
Glutathione Conjugate
(most of them: OATP2 substrate)
Rat MRP2 hMRP ○ ○ ○ ○ ○ ○ N.D. N.D. N.D. N.D. N.D. N.D. N.D. ○ N.D. N.D. N.D. N.D. ○ ○ ○ ○ ○ N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. ○ ○ ○ ○ N.D. ○ ○ ○ ○ ○ ○ ○ ○ N.D. ○ ○ ○ ○ N.D. N.D. ○ ○ ○ ○ ○ ○ ○ ○ ○ ○ ○ ○
ACE Inhibitors
temocaprilat
OATP2 MRP2
captopril enalaprilat etc.
Urine
Bile
Discrimination based on the difference in pharmacokinetic profile
(A)
(B) Cumulative biliary excretion amount (% of dose)
NH 2 out
in ABC2 ABC1
COOH
【Small Intestine】
【Blood-CSF Barrier】
CSF
P-gp/ MDR1 MRP2
BCRP
OATPs
PEPT1 ASBT
PepT2
Oatp3/ OATP-A
Oat3/ OAT3
Chproid plexus epithelium
Clinical Trial
Application Conventional Drug Development : 12 - 15 years Genomic Drug Development : 5 - 8 years
Oatp1 (670 a.a., M.W. 80kDa)
12 TMDs Tissue distribution: liver(sinusoidal membrane), kidney(brush border membrane), brain Substrates: organic anions:BSP, estradiol glucuronide, estrone sulfate, taurocholate, pravastatin, e tc organic cations:N-(4,4-azo-n-pentyl)-2 1-deoxyajamalinium, rocuronium
Plasma concentration (µg/ml)
10 EHBR
100
80
60 40
SD rat
1 SD rat
EHBR 20 0 0 2 4 Time (hr) 6
.1
0
1
2
3
4
5
6
Time (hr)
Plasma concentration (A) and cumulative biliary excretion (B) after i.v. administration of temocapril (1.0 mg/kg) in rats
blood
NTCP
OATP2/ OATP-C
OATP-B OAT2 OCT1 OATP OATPOATP8 8 B ATP
BSEP MRP2 bile
Jorg Konig et Am J Gastrointest Physiol Gastrointest Liver 2000 Am Jal, Physiol Liver Physiol Physiol ,278,G156-64,2000 Jan;278(1):G156-64