Table 1.
Workflow of high-throughput primary screening
| Step | Test compound concentration | Number of compounds | % of picked compounds | Inhibition cut-off (%) |
| Total library compound | - | 93,716 | - | - |
| Primary assay hits using FP | 20 µM | 3,010 | 3.21 | >75 |
| Secondary assay hits using AlphaLISA (AL) | 12.5 µM | 167 | 0.18 | >50 |
| Filtering using quenching (AL) | 12.5 µM | 97 | 0.1 | <30 |
| Lipinski’s rule | 87 | 0.09 | ≤1 violation | |
| Validation/dose–response Hits (FP and AL) | 0.31–40 µM | 26 | 0.028 | IC50 <50 µM |
| Dose–response using fresh compound (FP and AL) | 0.07–40 µM | 10 | 0.011 | IC50 <50 µM |