2-B-P-045 〇楠原 洋之、甲斐﨑 郁子、中島 佳子、戴 剣平 Hiroyuki Kusuhara, Ikuko Kaisaki, Yoshiko Nakajima, Kenpei Tai 東京大・院薬 Grad. Sch. of Pharm. Sci., Univ. of Tokyo [Purpose] The development of in vitro models that can quantitatively evaluate the bioavailability of drugs is an important issue in drug development. In this study, human iPS cell-derived intestinal epithelial cells (F-hiSIEC) were cultured on a chip manufactured by Emulate, a microperfusion device and evaluate their usefulness as a small intestine model for ADME study.
[Method] Emulate chips were coated with Matrigel and F-hiSIEC (2.4×105 cells/chip) were seeded. Cell culture was performed by stretching at 2% 0.15 Hz with flow rate of 30 μL/hr. Probe drugs were added to assess function of CYP3A4, P-gp and BCRP. The drug solution was collected from the outlet 6, 24, 30, and 36 hours after the start of perfusion.
[Results and Discussion] Formation of 1-OH form, which is a metabolite of midazolam produced by CYP3A4, was inhibited by ketoconazole, while it did not affect the midazolam cell permeability. Fg of midazolam in F-hiSIEC was estimated to be 0.97. Directional transport of quinidine, a P-gp substrate, was observed in the secretion direction (BtoA) over the absorption direction (AtoB) (ER 1.7~2.6). Similarly, sulfasalazine, a BCRP substrate, also showed in the secretory direction (ER 7). The permeability of antipyrine was not directional and affected by the inhibitors.
【Conclusion】 We succeeded in culturing F-hiSIEC on the Embed chip while maintaining the barrier function, CYP3A4, P-gp and BCRP functions. CYP3A4 activity is not sufficient to explain the metabolic capacity of in vivo, and correction using scaling factor is necessary.