Application of Intestinal Epithelial Cells Differentiated from Human Induced Pluripotent Stem Cells for Studies of Prodrug Hydrolysis and Drug Absorption in the Small Intestine

Abstract

Cell models to investigate intestinal absorption functions, such as those of transporters and metabolic enzymes, are essential for oral drug discovery and development. The purpose of this study was to generate intestinal epithelial cells from human induced pluripotent stem cells (hiPSC-IECs) and then clarify whether the functions of hydrolase and transporters in them reflect oral drug absorption in the small intestine. The hiPSC-IECs showed the transport activities of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), and peptide transporter 1 (PEPT1), revealed by using their probe substrates ([H]digoxin, sulfasalazine, and [C]glycylsarcosine), and the metabolic activities of CYP3A4, CES2, and CES1, which were clarified using their probe substrates (midazolam, irinotecan, and temocapril). The intrinsic clearance by hydrolysis of six ester prodrugs into the active form in hiPSC-IECs was correlated with the plasma exposure ( , AUC, and bioavailability) of the active form after oral administration of these prodrugs to rats. Also, the permeability coefficients of 14 drugs, containing two substrates of P-gp (doxorubicin and [H]digoxin), one substrate of BCRP (sulfasalazine), and 11 nonsubstrates of transporters (ganciclovir, [C]mannitol, famotidine, sulpiride, atenolol, furosemide, ranitidine, hydrochlorothiazide, acetaminophen, propranolol, and antipyrine) in hiPSC-IECs were correlated with their values of the fraction of intestinal absorption () in human clinical studies. These findings suggest that hiPSC-IECs would be a useful cell model to investigate the hydrolysis of ester prodrugs and to predict drug absorption in the small intestine.

Generation of Budding-Like Intestinal Organoids from Human Induced Pluripotent Stem Cells

“…25,27 Particularly, the expression levels of drug transporters and drug-metabolizing enzymes were low due to these immature characteristics; thus, they could not be used to study the pharmacokinetics of drugs. 39 Therefore, in this study, we attempted to generate intestinal organoids with characteristics similar to those of mature intestines that would be more useful for in vitro drug screening. We generated buddinglike organoids from human iPS cells through re-seeding on iMatrix 511-coated dishes and subsequent culturing with a medium containing the growth factor FGF2 and small molecular compounds A-83-01, CHIR99021, and Y-27632.…”

Direct On-Chip Differentiation of Intestinal Tubules from Induced Pluripotent Stem Cells

“…In-vitro models that closely mimic the human intestinal epithelium and its physiology are important tools for drug development processes [ 1 , 2 , 3 ]. As oral drug delivery is the preferred route of administration [ 4 ], such models have enormous potential to improve and speed up studies of absorption [ 5 ], disease modelling [ 6 ], drug metabolism and interactions as well as associated toxicity [ 7 , 8 ]. Therefore, patient-specific absorption models that take the gut complexity, passive and active transport and patient variability into consideration could be highly valuable in the evaluation of candidate drugs [ 9 ].…”

“…As opposed to human embryonic and adult stem cells, these cells are not associated with ethical concerns, they are easier to acquire and their generation does not involve the use of embryonic material [ 11 ]. Moreover, the ability of induced pluripotent stem cells to differentiate into the major cell types of the intestinal epithelium [ 5 , 12 , 13 ] makes them a useful tool to investigate the intestinal epithelium’s role in health and disease in-vitro. However, one of the major limitations of the organoid culture is the overall closed conformation, which makes the apical-luminal surface of the epithelium inaccessible [ 12 , 14 ] and, therefore, limits the applicability of these cultures for drug transport and absorption studies.…”

Inter‐individual and inter‐regional variations in enteric drug metabolizing enzyme activities: Results with cryopreserved human intestinal mucosal epithelia (CHIM) from the small intestines of 14 donors

“…In contrast, until recently, in vitro evaluation of human enteric drug metabolism has been hampered by the lack of experimental models with adequate drug metabolizing enzyme activities. Human in vitro enteric systems such as Caco‐2 and primary enterocyte cultures derived from crypt and stem cells are known to express drug metabolizing enzymes substantially lower than that in the human small intestine in vivo 17‐21 . To overcome this major obstacle in the evaluation of enteric drug metabolism, our laboratory evaluated an approach previously found successful with human in vitro hepatic models, namely isolation of metabolically competent cells from the organ of interest followed by immediate cryopreservation without culturing.…”