ABSTRACT
Purpose
Drug elimination in the liver consists of uptake, metabolism, biliary excretion, and sinusoidal efflux from the hepatocytes to the blood. We aimed to establish an accurate prediction method for liver clearance in rats, considering these four elimination processes. In vitro assays were combined to achieve improved predictions.
Methods
In vitro clearances for uptake, metabolism, biliary excretion and sinusoidal efflux were determined for 13 selected compounds with various physicochemical and pharmacokinetic properties. Suspended hepatocytes, liver microsomes and sandwich-cultured hepatocytes were evaluated as in vitro models. Based on the individual processes, in vivo hepatic clearance was calculated. Subsequently, the predicted clearances were compared with the corresponding in vivo values from literature.
Results
Using this in vitro-in vivo extrapolation method good linear correlation was observed between predicted and reported clearances. Linear regression analysis revealed much improved prediction for the novel method (r2 = 0.928) as compared to parameter analysis using hepatocyte uptake only (r2 = 0.600), microsomal metabolism only (r2 = 0.687) or overall hepatobiliary excretion in sandwich-cultured hepatocytes (r2 = 0.321).
Conclusions
In this new attempt to predict hepatic elimination under consideration of multiple clearance processes, in vivo hepatic clearances of 13 compounds in rats were well predicted using an IVIVE analysis method based on in vitro assays.
Abbreviations
- A:
-
total intracellular amount of radioactivity in sandwich-cultured hepatocytes
- ABC:
-
ATP binding cassette
- ATP:
-
adenosintriphosphate
- BCRP:
-
breast cancer resistance protein
- BDDCS:
-
biopharmaceutics drug disposition classification system
- BSEP:
-
bile salt export pump
- C:
-
total intracellular concentration of radioactivity in sandwich-cultured hepatocytes
- CLapp,met:
-
apparent metabolic clearance
- CLh:
-
hepatic plasma clearance
- CLh,int,in vitro :
-
hepatic intrinsic clearance predicted from in vitro assays
- CLh,org,in vitro :
-
hepatic organ clearance predicted from in vitro assays
- CLh,org,in vivo :
-
hepatic organ clearance based on blood concentrations
- CLmet:
-
metabolic clearance
- CLren:
-
renal clearance
- CLsandwich:
-
transcellular hepatobiliary clearance
- CLtot:
-
total organ plasma clearance
- DDI:
-
drug-drug interaction
- Er:
-
urinary excretion ratio
- fu(hep):
-
unbound fraction in hepatocytes
- fu(mic):
-
unbound fraction in liver microsomes
- fu,b:
-
unbound fraction in blood
- fu,p:
-
unbound fraction in plasma
- IVIVE:
-
in vitro-in vivo extrapolation
- kgbw:
-
kilogram body weight
- KHB:
-
Krebs-Henseleit buffer
- Km,bile:
-
affinity constant for the biliary excretion
- Km,efflux:
-
affinity constant for the active sinusoidal efflux
- Km,met:
-
metabolic affinity constant
- Km,uptake:
-
affinity constant for the active uptake
- LC-MS:
-
liquid chromatography coupled with mass spectrometry
- logD:
-
distribution coefficient
- LOQ:
-
limit of quantification
- LSC:
-
liquid scintillation counting
- MDR/Mdr:
-
multi-drug resistance protein
- mRNA:
-
messenger ribonucleic acid
- MRP/Mrp:
-
multi-drug resistance associated protein
- N.A.:
-
not applicable
- N.D.:
-
not determined
- OATP/Oatp:
-
organic anion transporting polypeptide
- PBPK:
-
physiologically based pharmacokinetics
- PSapp,bile:
-
apparent biliary clearance
- PSbile:
-
biliary clearance
- PSefflux,active:
-
apparent sinusoidal active efflux clearance
- PSefflux,passive:
-
sinusoidal passive efflux clearance
- PSefflux,total:
-
apparent sinusoidal total efflux clearance
- PSuptake,active:
-
apparent active uptake clearance
- PSuptake,passive:
-
non-specific passive diffusion
- PSuptake,total:
-
total apparent uptake clearance
- Qh:
-
rat hepatic blood flow rate
- R:
-
compound amount in bile pocket
- radio-HPLC:
-
high performance liquid chromatography coupled with on-line radio detection
- Rb:
-
blood-to-plasma concentration ratio
- S:
-
nominal incubation concentration in buffer system
- t:
-
incubation time
- Vapp,met:
-
apparent metabolic velocity
- Vc:
-
rat hepatocyte volume
- Vmax,bile:
-
maximum velocity for the biliary excretion
- Vmax,efflux:
-
maximum velocity for the active sinusoidal efflux
- Vmax,met:
-
maximum metabolic velocity
- Vmax,uptake:
-
maximum velocity for the active sinusoidal uptake
- Vuptake,total:
-
total apparent uptake velocity
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ACKNOWLEDGMENTS & DISCLOSURES
The authors wish to acknowledge the many Novartis Drug Metabolism and Pharmacokinetic Department Scientists of Basel Switzerland who have supported generation of data used in these analyses. Special thanks go to Drs. Heike Gutmann, Joel Krauser and Birk Poller for their critical evaluation of this work.
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Umehara, Ki., Camenisch, G. Novel In Vitro-In Vivo Extrapolation (IVIVE) Method to Predict Hepatic Organ Clearance in Rat. Pharm Res 29, 603–617 (2012). https://doi.org/10.1007/s11095-011-0607-2
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DOI: https://doi.org/10.1007/s11095-011-0607-2