A Novel Plated Hepatocyte Relay Assay (PHRA) for In Vitro Evaluation of Hepatic Metabolic Clearance of Slowly Metabolized Compounds

Title:A Novel Plated Hepatocyte Relay Assay (PHRA) for In Vitro Evaluation of Hepatic Metabolic Clearance of Slowly Metabolized Compounds

Volume: 10 Issue: 1

Author(s): Chi-Chi Peng, Utkarsh Doshi, Chandra Prakash and Albert P. Li

Affiliation:

Keywords: Cryopreserved hepatocytes, hepatic clearance, human hepatocytes, low clearance compounds, metabolic clearance, relay assay, slowly metabolized compounds.

Abstract: Objective: Development and validation of a novel assay, the Plated Hepatocyte Relay Assay (PHRA), for the determination of the metabolic fates of slowly metabolized compounds. Method: Cryopreserved human hepatocytes were cultured for 4 h followed by incubation with slowly metabolized compounds for 24 h (initial incubation). On the next day, the incubated media were collected and added to hepatocytes was similarly prepared on the day of incubation (48 h incubation; 1^st relay). The procedures were repeated on the next days (72 h (2^nd relay), 96 h (3rd relay), and 120 h (4^th relay) incubations). Results: A proof-of-concept study with two low clearance compounds, diazepam and tolbutamide, and a validation study with 15 ultra-low clearance compounds (CL_non-renal < 1 mL/min/kg) and low clearance compounds (CL_non-renal 1- 5.1 mL/min/kg) were performed. Linear time-dependent disappearance of the parent compounds was observed for all compounds. Application of published free fraction values in combination with a correction factor with in vitro hepatic clearance results obtained with the PHRA accurately predicted in vivo hepatic clearance. Conclusion: PHRA represents a useful experimental system for the evaluation of the metabolic fates of low clearance compounds in drug development..  

Cite this article as:

Peng Chi-Chi, Doshi Utkarsh, Prakash Chandra and Li P. Albert, A Novel Plated Hepatocyte Relay Assay (PHRA) for In Vitro Evaluation of Hepatic Metabolic Clearance of Slowly Metabolized Compounds , Drug Metabolism Letters 2016; 10 (1) . https://dx.doi.org/10.2174/1872312809666150818111500