Method Profile
Combine a donor-derived intestinal epithelial barrier with transporter, metabolic and mathematical absorption analyses.
Measure oral-drug permeability and intestinal metabolism and assess conditional extrapolation to human Fa or FaFg.
Duodenal crypt-derived human organoids from surgical adjacent-normal tissue; seven donors in culture optimization and four in drug-permeability comparisons.
Transwell monolayers; four-day expansion and six-day differentiation with lipid-rich albumin and final-two-day 100 nM 1α,25-dihydroxyvitamin D3 (VD3); FaSSIF-compatible permeability assays.
Barrier and metabolic culture
Organoid-derived epithelium on laminin-fragment/fibronectin coating; retained EGF during differentiation; AlbuMAX supports formation across donors.
Functional probes
Lucifer yellow for barrier integrity; digoxin/zosuquidar and sulfasalazine/Ko143 for efflux; midazolam with or without CYP inhibition or rifampicin induction; separate UGT and CES2 probes.
Use LC–MS/MS permeability data and explicit compartment assumptions. Distinguish fitted permeability–Fa correlation from FaFg estimates that combine inhibitor data and a CYP3A expression-based activity scaling assumption.
Stepwise culture optimization, donor/repeat comparisons and retrospective comparison with marketed-drug human absorption data.
Seven donor cultures in optimization; four donors for 19-drug profiles; donor-1 data for 19-drug Fa and nine-drug FaFg estimates. Barrier CV comparison used n = 24 biological replicates.
Alternative culture conditions, primary enterocyte expression, probe inhibitors/induction and reported human Fa or FaFg.
Fa correlation R² = 0.88 for 19 drugs after regression fitting. CYP-inhibitor-based FaFg calculation placed three of nine drugs within 1.5-fold; expression-scaled calculation placed seven of nine within 1.5-fold. Rifabutin was overestimated by every approach. Barrier-assay CV was 8.6% in the n = 24 comparison.
A Chugai-led study optimized organoid-derived intestinal monolayers across seven donors. Permeability correlated with human absorption for 19 marketed drugs, while estimates for metabolized drugs depended on an explicit expression-based CYP3A scaling assumption.
