Table 4. Cell and Counterscreening Assay Data for Selected Compounds.
| compound | HSET ADP-Glo IC50 (μM)a | HSET ADP-Glo (ATP 500 μM) IC50 (μM)b | Eg5 ADP-Glo IC50 (μM)c,e | % multipolarity above baseline @ 15 μM (4NCA cells)f | % multipolarity above baseline @ 15 μM (4N cells)g |
|---|---|---|---|---|---|
| 1 | 0.171 (±0.10) | 0.825d | 35.6 (36.4, 34.9) | ||
| 2 | 2.73 (±0.71) | >200 | 0% | 1% | |
| 13 | 0.063 (±0.017) | >200 | 9% | 1% | |
| 18 | 0.027 (±0.007) | 0.086c | >200 | 11% | 0% |
| 26 | 0.093 (±0.041) | 0.332c | >200 | 13% | 1% |
| 32 | 0.012 (±0.006) | 0.051c | >200 | 21% | 2% |
| 35 | 0.019 (±0.007) | 0.066c | >200 | 11% | 1% |
| 36 | 0.011 (±0.004) | 0.090d | >200 | 15% | 1% |
Inhibition of recombinant full-length HSET with preformed microtubules and 3 μM ATP measured in ADP-Glo format, mean (±SD) for n ≥ 3.
Inhibition of recombinant full-length HSET with preformed microtubules and a high ATP concentration of 500 μM measured in ADP-Glo format.
Single determination.
Mean of two results.
Inhibition of commercially available GST-tagged Eg5 kinesin with preformed microtubules and 4.8 μM ATP measured in ADP-Glo format.
Multipolar spindle assay (4NCA cell line). The percentage of multipolar mitoses was calculated by dividing the number of multipolar mitoses by the total number of all visible mitoses in one well of a 96-well plate (n > 100 for each replicate, 2 replicates per concentration point).
Multipolar spindle assay (4N cell line). The percentage of multipolar mitoses was calculated by dividing the number of multipolar mitoses by the total number of all visible mitoses in one well of a 96-well plate (n > 100 for each replicate, 2 replicates per concentration point).