BioCentury
ARTICLE | Distillery Techniques

Disease models; drug properties

April 17, 2018 9:13 PM UTC

3-D human intestinal organoids could be used to test the PK properties of drug compounds. The organoids are generated by culturing human induced pluripotent stem (iPS) cells in a medium containing fibroblast growth factor 4 (FGF4) and a glycogen synthase kinase 3 (GSK3) inhibitor; seeding the cells in commercially available low-adherence scaffolds; suspending them in culture containing Matrigel and epidermal growth factor (EGF) to generate uniform spheroids; and differentiating them into intestinal tissue using inhibitors of DNA methyltransferase and γ secretase. The resulting intestinal organoids contain multiple intestinal cell types and express high levels of mRNA for cytochrome P450 3A4 (CYP3A4), the tight junction protein occludin (OCLN), and the ATP-binding cassette sub-family B member 1 (ABCB1; MDR1; PGP; CD243) efflux pump. In the intestinal organoids, the generic antibiotic rifampicin and a biologically active form of vitamin D3 -- both of which induce expression of intestinal CYP3A4 -- increased CYP3A4 mRNA levels and CYP3A4 metabolic activity compared with vehicle. Incubation of the organoids with the CYP3A4 substrate midazolam increased levels of midazolam metabolites compared with vehicle, and co-incubation with midazolam and the CYP3A4 inhibitor ketoconazole decreased levels of midazolam metabolites by about 38%. Next steps could include using the method to test the PK properties of other drug compounds.

The generic midazolam, a benzodiazepine modulator of the GABA receptor, is marketed for anesthesia, seizures and insomnia...