Evaluation of drug-drug interactions at the level of hepatic excretion
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Transcript of Evaluation of drug-drug interactions at the level of hepatic excretion
Evaluation of drug-drug interactions at the level of hepatic excretion
Stanislav Mičuda
Department of Pharmacology, Charles University in PragueFaculty of Medicine in Hradec Králové
55. Česko-Slovenské Farmakologické dny30.8.-1.9.2005, Hradec Králové
55. Farmakologické dnyHepatic
drug processing
MDR1
MDR3
NTCP
MRP2 MRP2
NTCP
BCRP
OCT1
MRP3
Phase IPhase II
BILE SALTS BULKY ORGANIC
COMPOUNDSORGANICANIONS
SMALL ORGANICCATIONS
OATs
BSEP
OATP2
Inhibition – MDR1
Shon JH; Clin Pharmacol Ther 2005;78(2):191-201 Kovarik JM; Clin Pharmacol Ther 1999;66(4):391-400
Itraconazole
Digoxin 0.125 mg +valspodar 400 mg 7-11 day
Greiner B; J Clin Invest 1999;104(2):147-53.
Induction – MDR1
In vivo
Single and Repeat Dose PharmacokineticsDrug Interaction StudiesMass Balance Excretion Studies Imaging StudiesIn Situ Tissue Perfusions
Preclinical -Clinical Studies
Mai I; Clin Pharmacol Ther 2004;76(4):330-40.Sasongko L; Clin Pharmacol Ther 2005;77(6):503-14.
99mTc-Sestamibi Scan following XR-9576
– a synthetic corticosteroid, (t1/2 = 36-54 h), eliminated mainly in the unchanged form in urine – induction of mrp2 protein expression?
SPC: 86698 DEXAMETHAZON LÉČIVA TBL 20X0.5MG-BLISTR LEX CZ
Dexamethasone
The aim of study
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influence of dexamethasone on expression and activity of mrp2
Methotrexate (MTX) as a selective substrate for mrp2
Probenecid as an inhibitor of mrp2
In vivo pharmacokinetics
Wistar male rats (Konárovice) - 3 groups (N = 6) Dex (25 mg/kg daily 4 days, p.o.),Control (Oliv. oil 4 days, p.o.), Probenecid 70 mol/kg 3.33 mol/min.kg-1
simultaneously with MTX
Clearance study left jugular v. (MTX 22 mol/kg 164 nmol/kg.h-1 4 ml/min) right a. carotis (blood sampling - since 55‘ - 10 min) bladder (urine 0-90 min 10 min) bile duct (bile 0-90 min 10 min)
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MTX concentrations HPLC method with fluorescent detection *
Farmacokinetic analysis
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In vivo pharmacokinetics
CLtotal = Rinf / Css
CLbil = Cbil . Vbil / Css
CLren = CU.VU / Css
BE = Cbil . Vbil
UE = CU.VU
CLRratio = CLren / GFR
CLRratioU = CLren / (fU .GFR)
CLRS = (CLren / fU) - GFR
* Chladek J; J Chromatogr B Biomed Sci Appl 2000;744(2):307-13.
Pharmacokinetics of MTX in steady-state
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60 70 80 90 100
0
10
20
30
40Probenecid
Control
Dexamethasone
Time (min)
Me
tho
tre
xa
te (
uM
)
Pharmacokinetics of MTX in steady-state
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Control DEX 25 mg po Probenecid DIF
(fold) values DIF
(fold) Urine flow rate (l/min) 28.9 4.7 71.4 8.2 C 2.5 27.7 8.5 1.0
UE rate (nmol/ml.kg-1) 92.7 10.6
85.9 5.8 0.9 57.0 13.5 A 0.6
Bile flow rate (l/min) 25.2 1.6 28.0 2.7 1.1 32.7 2.8 A 1.3
BE rate (nmol/ml.kg-1) 98.5 8.2 124.0 9.9 A 1.3 62.0 5.2 B 0.6
Plasma (M) 18.4 1.3 20.2 2.2 1.1 35.4 1.2 C 1.9
CLR (ml/min.kg-1) 5.0 0.4 4.6 0.7 0.9 1.6 0.4 C 0.3
CLBile (ml/min.kg-1) 5.6 0.7 6.4 0.6 1.1 1.8 0.2 C 0.3
CLTOT (ml/min.kg-1) 9.2 0.6 8.7 1.0 0.9 4.7 0.2 C 0.5
CLCR (ml/min.kg-1) 4.8 0.6 4.9 0.4 0.9 3.6 0.4 0.7
fU 0.5 0.02 0.7 0.1 + 1.3 0.6 0.03 B 1.3
CLR/fu 10.0 0.2 6.7 0.2 B 0.7 2.4 0.2 C 0.2
CLR/GFR 1.1 0.9 1.1 0.4 0.9 0.5 0.5 B 0.4
CLRS 5.3 1.0 2.2 0.6 A 0.4 -1.2 0.8 C -0.2
CLR/(fU x GFR) 2.2 0.3 1.6 0.2 + 0.7 0.7 0.2 B 0.4
Values are means SEM A P < 0.05; B P < 0.01; C P < 0.001
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Dexametazon 25 mg/kgOlive oilRat liver - mrp2 – Dex per os
Control Dexamethasone Parameter for image analysis
Centr Port Centr Port
Proportion of visual field area occupied by mrp2 staining
0.13 ± 0.05 2.11 ± 0.772 2.92 ± 0.88 C 3.91 ± 1.14 A
Integral optical densities of mrp2 in visual field
28 ± 11 467 ± 1742 664 ± 202 C 903 ± 266 A
Values are means ± SEM of 6 rats. Centr - pericentral areas; Port - periportal areas; 1P<0.05, 2P<0.001 periportal vs pericentral areas; AP<0.05, CP<0.001 comparison of data from respective locations between control and dexamethasone (25 mg/kg daily, p.o. for 4 days) pretreated animals.
BE 1.3-foldCLbile 1.1-foldHistol 3.1-foldWestern 2.4-fold
Pharmacokinetics of Rho-123
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Influence of DEXon P-gp activity
Control DEX 25 mg po
DIF (fold)
Urine flow rate (l/min) 26.0 4.0 96.0 12.0 C 3.6
UE rate (nmol/ml.kg-1) 0.10 0.04 0.28 0.06 B 2.7
Bile flow rate (l/min) 27.0 2.0 28.0 4.0 1.0
BE rate (nmol/ml.kg-1) 0.26 0.04 0.57 0.14 A 2.2
Plasma (M) 0.12 0.02 0.08 0.003 B 0.6
CLR (ml/min.kg-1) 0.71 0.24 3.70 0.68 B 5.2
CLBile (ml/min.kg-1) 2.22 0.34 7.88 2.29 A 3.5
CLTOT (ml/min.kg-1) 21.68 1.00 29.26 1.18 B 1.4
CLCR (ml/min.kg-1) 1.19 1.01 1.34 1.20 1.1
CLR/GFR 0.59 0.08 2.90 0.10 B 4.9
Values are means SEM A P < 0.05; B P < 0.01; C P < 0.001 Ctrl DEX
BE 2.2-foldCLbile 3.5-foldHistol 2.3-foldWestern 2.8-foldReal RT-PCR 3.1-fold
Conclusion
Drug-drug interactions may be of great clinical importance - A significant number is transport protein-mediated- in vivo studies – a complex view of pharmacokinetics
and mechanisms using model substrates
- in vivo studies – verification of outcome from in vitro models
allowing predictions- interspecies extrapolation
Glucocorticoids and MRP2 vs. MTX- induction of proteins without changes in pharmacokinetics of MTX
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ACKNOWLEDGEMENTS
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Grants GAUK 89/2002/CMŠMT EC-B25.001
Co-workersLeoš FuksaJolana CermanováEva Brčáková Jitka Hájková František ŠtaudPetr Pávek Lucie MundlováJan OsterreicherJaroslav MokrýJaroslav ChládekJiřina Martínková
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