BCRP Inhibition assay (Caco-2 cells)
Aim: To determine the inhibition potential of test compound for BCRP (Breast Cancer Resistance Protein Multidrug Transporter), an efflux transporter, using the Caco-2 cell model.
Background information: Caco-2 cells are a human colon adenocarcinoma cell line which express efflux transporters, such as P-gp (P-glycoprotein) and BCRP. Efflux transporters are involved in drug absorption and disposition. Inhibition of transporter-mediated efflux may affect the efficacy and safety of potential concomitant drugs in a clinical setting.
Inhibition transport assays are conducted both in the absorption (apical-to-basolateral) and efflux (basolateral-to-apical) directions. Probe substrate is added to either the apical or basolateral sides of the monolayer in the absence and presence of test compound. Probe substrate concentrations are quantified by UPLC-MS/MS using a calibration curve.
Compound requirement: 5 mg
Turnaround time: 4 weeks
- Test compound concentrations: 3 (variable on request and compound solubility)
- 24-well plate
- Number of replicates: 2
- DMSO concentration: 1%
- Incubation time: 120 min
- Probe substrate: nitrofurantoin
- Positive control: Ko143
- Monolayer integrity marker: lucifer yellow
- Analytical method: UPLC-MS/MS with calibration curves
- Absorption and efflux permeability of the probe substrate (+/- test compound)
- Absorption and efflux recovery of the probe substrate (+/- test compound)
- Efflux ratio of the probe substrate (+/- test compound)
C1, C2, C3: test compound concentrations
NIT: Nitrofurantoin (probe substrate)
Ko: Ko143 (inhibitor)