Data Supplement
- Data Supplement -
Supplemental Methods
Supplemental Table 1 - Suppliers of materials for in vitro studies.
Supplemental Table 2 - LC methods for measurement of darolutamide and metabolites in plasma, urine and feces.
Supplemental Table 3 - Range of accuracy and precision data of LC-MS/MS methods for (S,R)-darolutamide, (S,S) darolutamide, and keto-darolutamide across all validation quality control replicates.
Supplemental Table 4 - LC-MS methods for quantitation of darolutamide and metabolites in vitro.
Supplemental Table 5 - NMR data of metabolites M-7a, M-15a, M-21 and M-22 isolated from pooled human urine
Supplemental Table 6 - Molecular ions and characteristic fragment ions of darolutamide and metabolites detected in biological samples.
Supplemental Table 7 - Depletion of 14C-darolutamide in human hepatocytes of two donors (HH-TZU, HH-TWT) in the absence and presence of the CYP3A4 inhibitor itraconazole for up to 60 minutes incubation time.
Supplemental Table 8 - Correlations between UGT isoform-selective activity and formation of O- and N-glucuronides in human liver microsomes.
Supplemental Table 9 - Contribution of UGT1A1 and UGT1A9 to M-7a and M-7b formation calculated based on experiments with chemical inhibitors and by relative activity factors (RAF).
Supplemental Figure 1 - HPLC pattern of 14C-keto-darolutamide stock solution before (A) and after (B) incubation for 24 hours at 37°C with human feces at a concentration of 2 μM.
Supplemental Figure 2 - Oxidation of 1 μM (S,R)-darolutamide (A) and 1μM (S,S)- darolutamide (B) to keto-darolutamide in human liver cytosol in the presence of cofactors (NAD, NADP).
Supplemental Figure 3 - Depletion of 14C-keto-darolutamide (1 μM) after 1-hour incubation with a complete panel of all available recombinant CYP enzymes or human liver microsomes.
Supplemental Figure 4 - Formation of glucuronides M-7a/b and M-15a/b catalyzed by various recombinant human UGT isoforms applying 1 μM darolutamide (A), (S,R)- darolutamide (B), and (S,S)-darolutamide (C).
Supplemental References