Title:A Novel In vitro Experimental System for the Evaluation of Enteric Drug Metabolism: Cofactor-Supplemented Permeabilized Cryopreserved Human Enterocytes (MetMax™ Cryopreserved Human Enterocytes)
VOLUME: 12 ISSUE: 2
Author(s):Albert P. Li*, Kirsten Amaral and Ming-Chih D. Ho
Affiliation:In vitro ADMET Laboratories Inc., 9221 Rumsey Road, Suite 8, Malden, MA 02148, USA and Boston Hepatocyte Technology Center, In vitro ADMET Laboratories, 389 Main St, Ste 301, Malden, MA 02148, In vitro ADMET Laboratories Inc., 9221 Rumsey Road, Suite 8, Malden, MA 02148, USA and Boston Hepatocyte Technology Center, In vitro ADMET Laboratories, 389 Main St, Ste 301, Malden, MA 02148, In vitro ADMET Laboratories Inc., 9221 Rumsey Road, Suite 8, Malden, MA 02148, USA and Boston Hepatocyte Technology Center, In vitro ADMET Laboratories, 389 Main St, Ste 301, Malden, MA 02148
Keywords:Permeabilized, cryopreserved human enterocytes, MMHE, CCHE, Drug metabolism enzyme (DME), enteric drug
metabolism.
Abstract:Background: We report here an evaluation of a novel experimental system- cofactorsupplemented
permeabilized cryopreserved human enterocytes (MetMax™ cryopreserved human enterocytes
(MMHE), patent pending) for applications in the evaluation of enteric drug metabolism. A
major advantage of MMHE over Conventional Cryopreserved Human Enterocytes (CCHE) is the simplification
of the use procedures including storage at -80°C instead of in liquid nitrogen, and use of the
cells immediately after thawing without a need for centrifugation and microscopic evaluation of cell
density and viability and cell density adjustment.
Methods: In this study, we compared MMHE and CCHE in key phase 1 oxidation and phase 2 conjugation
Drug Metabolism Enzyme (DME) activities that we recently reported for cryopreserved human
enterocytes: 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 all
the DME pathways evaluated, with specific activities of MMHE ranged from 142% (CYP2C9) to
1713% (UGT) of that for CCHE. β-hydroxylation and testosterone 6.
Result and Conclusion: Our results suggest that the MMHE system represents a convenient and robust
in vitro experimental system for the evaluation of enteric drug metabolism.
