Table 2.
Assays used for determining IDILI potential.
| Mechanisms | Proteins | Relevant assays |
|---|---|---|
| Phase I DMEs | CYP450 | Coumarin metabolism Fluorescein (CYP2C9) Human liver microsomes (HLMs) High pressure liquid chromatography (HPLC)/Mass spectrometry (MS) |
|
| ||
| Phase II DMEs | UGT | Coumarin metabolism |
| GST (GSTT1 and GSTM1) | CDNB, NBC, DCNB, EPNP, DCM, PBO GSTT1/GSTM1 single and double knockouts |
|
| UGT/GST/NAT/SULT | Covalent binding level (CBL) HPLC/MS |
|
|
| ||
| Antioxidant enzymes | GPx1/SOD2 | Monochlorobimane (mBCl) Oxygen consumption rate (OCR) Fluorescein Tetramethyl rhodamine (TMRM) Cytochrome c release SOD2+/- mice |
|
| ||
| Transporters | BSEP | Vesicular transport assay (VTA) ATPase assay |
| MRP2 | Fluorescein | |
| MDR1/MRP2 | Flow cytometry | |
| OCT | OCT1 and OCT2 knockouts | |
| BSEP/BCRP/MDR/MRP/OATP/OCT/NTCP | HPLC/MS Drug uptake assays |
|
|
| ||
| Inflammation | TNFR1/IFN-γ/TLR9/IL-1/IL-4/IL-6/IL-10/IL-13 | LPS-induced inflammation Lymphocyte transformation test (LTT) Cytokine production Cytokine knockouts |
| CD69 | Flow cytometry | |
|
| ||
| HLA | MHC I/MHC II | Flow cytometry |
Cytochrome P450 (CYP450), uridine 5′-diphosphate (UDP) glycosyltransferase (UGT), glutathione S-transferase (GST), N-acetyl transferase (NAT), sulfotransferase (SULT), glutathione peroxidase (GPx), superoxide dismutase (SOD), 1-chloro-2,4-dinitrobenzene (CDNB), p-nitrobenzyl chloride (NBC), 1,2-chloro-4-nitrobenzene (DCNB), 2-epoxy-3-(p-nitrophenoxy)propane (EPNP), dichloromethane (DCM), trans-4-phenyl-3-buten-2-one (PBO), lipopolysaccharide (LPS), tumor necrosis factor R1 (TNFR1), interferon gamma (IFN-γ), toll-like receptor (TLR), interleukin (IL), cluster of differentiation (CD), major histocompatibility complex class I (MHC I), and class II (MHC II).