Prediction of hepatic clearance using cryopreserved human hepatocytes: a comparison of serum and serum-free incubations.
Cryopreserved human hepatocytes have been used to predict hepatic in-vivo clearance. Physiologically-based direct scaling methods generally underestimate human in-vivo hepatic clearance. Cryopreserved human hepatocytes were incubated in 100% serum and in serum-free medium to predict the in-vivo hepatic clearance of six compounds (phenazone (antipyrine), bosentan, mibefradil, midazolam, naloxone and oxazepam). Monte Carlo simulations were performed in an attempt to incorporate the variability and uncertainty in the measured parameters to the prediction of hepatic clearance. The intrinsic clearance (CL(int)) and the associated variability of the six compounds decreased in the presence of serum and the values were reproducible across donors. The predicted CL(hep, in-vivo) obtained with hepatocytes from donors incubated in serum was more accurate than the prediction obtained in the absence of serum. For example, the CL(hep, in-vivo) of mibefradil in donor GNG was 4.27 mL min(-1) kg(-1) in the presence of serum and 0.46 mL min(-1) kg(-1) in the absence of serum (4.88 mL min(-1) kg(-1) observed in-vivo). Using the results obtained in this study together with an extended data set (26 compounds), the clearance of 77% of the compounds was predicted within a 2-fold error in the absence of serum. In the presence of serum, 85% of the compounds were successfully predicted within a 2-fold error. In conclusion, cryopreserved human hepatocyte suspensions represented a convenient and predictive model to assess human drug clearance.
Blanchard
Nadège
N
F. Hoffmann-La Roche AG, Pharmaceuticals Division, CH-4070 Basel, Switzerland.
Hewitt
Nicola J
NJ
Silber
Paul
P
Jones
Hannah
H
Coassolo
Philippe
P
Lavé
Thierry
T
D000983:Antipyrine / Q000378:metabolism; D000077300:Bosentan; D018929:Cell Culture Techniques; D015925:Cryopreservation; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008657:Metabolic Clearance Rate; D008954:Models, Biological; D009010:Monte Carlo Method; D010076:Oxazepam / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011485:Protein Binding; D015203:Reproducibility of Results; D044967:Serum / Q000378:metabolism*; D013449:Sulfonamides / Q000378:metabolism
D004364:Pharmaceutical Preparations; D013449:Sulfonamides; D010076:Oxazepam; D000077300:Bosentan; D000983:Antipyrine