A Novel In vitro Experimental System for the Evaluation of Enteric Drug Metabolism: Cofactor-Supplemented Permeabilized Cryopreserved Human Enterocytes (MetMax? Cryopreserved Human Enterocytes)

摘要

Background: We report here an evaluation of a novel experimental system- cofactorsupplementedpermeabilized cryopreserved human enterocytes (MetMax? cryopreserved human enterocytes(MMHE), patent pending) for applications in the evaluation of enteric drug metabolism. Amajor advantage of MMHE over Conventional Cryopreserved Human Enterocytes (CCHE) is the simplificationof the use procedures including storage at -80°C instead of in liquid nitrogen, and use of thecells immediately after thawing without a need for centrifugation and microscopic evaluation of celldensity and viability and cell density adjustment.Methods: In this study, we compared MMHE and CCHE in key phase 1 oxidation and phase 2 conjugationDrug Metabolism Enzyme (DME) activities that we recently reported for cryopreserved humanenterocytes: CYP2C9 (diclofenac 4’- hydroxylation), CYP2C19 (s-mephenytoin hydroxylation),CYP3A4 (midazolam 1’-hydroxylation), CYP2J2 (astemizole O-demethylation), uridine 5'-diphosphoglucuronosyltransferase(UGT; 7-hydroxycoumarin glucuronidation), sulfotransferase (SULT; 7-hydroxycoumarin sulfation), N-acetyl transferase-1 (NAT-1; p-benzoic acid N-acetylation), and carboxyesterase-2 (CES-2; hydrolysis of irinotecan to SN38). Both CCHE and MMHE were active in allthe DME pathways evaluated, with specific activities of MMHE ranged from 142% (CYP2C9) to1713% (UGT) of that for CCHE. β-hydroxylation and testosterone 6.Result and Conclusion: Our results suggest that the MMHE system represents a convenient and robustin vitro experimental system for the evaluation of enteric drug metabolism.