Summary of probe substrate metabolic reactions used in in vitro drug metabolism studies
P450 | Preferred Substrate Reaction | Km | Recommended Organic Solvent2-a |
---|---|---|---|
μM | <1%, v/v | ||
1A2 | PhenancetinO-deethylation | 10–50 | DMSO, methanol |
2A6 | Coumarin 7-hydroxylation | 0.5–2 | DMSO, methanol |
2B6 | S-mephenytoinN-demethylation | 1900 | N.R. |
2C9 | Tolbutamine 4′-hydroxylation | 100–200 | Acetonitrile |
S-Warfarin 7′-hydroxylation | 1–5 | N.R. | |
2C19 | S-mephenytoin 4′-hydroxylation | 31–340 | Acetonitrile, methanol |
2D6 | Bufuralol 1′-hydroxylation | 4–10 | Acetonitrile |
DextromethorphanO-demethylation | 2–10 | DMSO, acetonitrile, methanol | |
2E1 | Chlorzoxazone 6-hydroxylation | 40 | Acetonitrile |
3A4/5 | Testosterone 6β-hydroxylation | 50–100 | Acetonitrile, methanol |
Midazolam 1′-hydroxylation | 3–5 | N.R. |
N.R., not reported.
↵2-a Effect less than 20% is deemed acceptable. Recommendation is based on at least two consistent experimental results available in the studies by Draper et al., (1997), Chauret et al., (1998), Hickman et al., (1998), Busby et al., (1999), orTang et al., (2000), with the exception of bufuralol 1′-hydroxylation in incubation with acetonitrile, which is only reported by Busby et al. (1999) in cDNA-expressed enzymes.