Accurately determine the intrinsic clearance for metabolically stable compounds for which a traditional suspension assay fails to quantify.
The low clearance hepatocyte stability assay is in our portfolio of in vitro ADME services. We deliver consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.
Optimization of clearance is one of the more significant challenges for a drug discovery project. Identification of the rate in preclinical species and optimization in human are major goals in most projects
1Grime KH, Barton P & McGinnity DF (2013) Mol Pharm 10; 1191-1206
Cells | Primary human hepatocytes |
---|---|
Test Compound Concentration | 1 µM (different concentrations available) |
Overlay Matrix | Geltrex® |
Incubation Time | 0, 1, 2, 4, 8, 22, 26, 30 h |
Replicates | n=2 |
Compounds Requirements | 20 μL of 10 mM solution |
Analysis Method | LC-MS/MS quantification |
Assay Controls | Disopyramide (low clearance) Metoprolol (moderate clearance) Sildenafil (high clearance) |
Data Delivery | Intrinsic clearance Half life |
Ion Class | Major Drug Metabolising Enzyme | Bonn et al., 2016 PHH CLint (µL/min/106 cells) | Bonn et al., 2016 Hurel CLint (µL/min/106 cells) | Evotec CLint (µL/min/106 cells) | |
---|---|---|---|---|---|
Bupropion | Base | CYP2B6, CYP1A2, CYP2A6, CYP3A4, CYP2E1 | Not reported | Not reported | 5.4 |
Carvedilol | Base | CYP2D6, CYP2C9 | 26.3 | 34.2 | 14.5 |
Diazepam | Neutral | CYP2C19, CYP3A4 | 0.8 | 1.3 | 0.7 |
Diclofenac | Acid | CYP2C9, UGT2B7 | Not reported | Not reported | 4.7 |
Disopyramide | Base | CYP3A4 | 0.2 | 0.4 | 0.1 |
Ethinylestradiol | Acid | UGT1A1, CYP3A4 | Not reported | Not reported | 3.3 |
Imipramine | Base | CYP1A2, CYP2C19, CYP2D6 | 8.6 | 1.7 | 8.5 |
Metoprolol | Base | CYP2D6, CYP3A4 | 2.2 | 0.8 | 0.9 |
Midazolam | Neutral | CYP3A4 | Not reported | Not reported | 5.1 |
Sildenafil | Base | CYP3A4, CYP2C9, CYP2C19 | 7.0 | 6.2 | 9.0 |
Tolbutamide | Acid | CYP2C9 | Not reported | Not reported | 0.8 |
Warfarin | Neutral | CYP2C9, CYP3A4 | BLQ | 0.7 | 0.3 |
1Grime KH et al., (2013) Application of in silico, in vitro and preclinical pharmacokinetic data for the effective and efficient prediction of human pharmacokinetics. Mol Pharm 10(4); 1191-1206
2 Bonn B et al. (2016) Determination of human hepatocyte intrinsic clearance for slowly metabolised compounds: Comparison of a primary hepatocyte/stromal cell co-culture with plated primary hepatocytes and HepaRG. Drug Metab Dispos 44; 527-533
Learn more about drug metabolism in Chapter 3 of our popular Everything you need to know about ADME guide.
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