Table 1.
Comparison of a Single-Point Luciferase Screen in Rabbit Reticulocyte Lysate Compared with the Cell-Based Luciferase Assay
| Compound | RRL (% inhibition) | HCT116 (% inhibition) | PC3-MM2 (% inhibition) | A549 (% inhibition) |
|---|---|---|---|---|
| DMSO | 0 | 0 | 0 | 0 |
| 17-AAG | 81±9 | 51±8 | 46±6 | 46±8 |
| 36 | 43±3 | 34±45 | −1±25 | 6±4 |
| 111 | 55±5 | 36±24 | 9±13 | −2±9 |
| 130 | 44±5 | 4±6 | −2±21 | −11±16 |
| 131 | 38±6 | −2±8 | −5±15 | −10±16 |
| 133 | 39±16 | −2±16 | −6±12 | −9±16 |
| 174 | 72±1 | 51±20 | 40±25 | 44±15 |
| 252 | 83±14 | 45±16 | 30±27 | −4±6 |
| 298 | 55±0 | −13±69 | −16±19 | −7±9 |
| 390 | 64±0 | 6±2 | 6±12 | −19±18 |
| 401 | 78±2 | 90±7 | 52±14 | 51±29 |
| 403 | 72±1 | 61±13 | 34±15 | 35±23 |
| 404 | 56±1 | 14±3 | 26±15 | −1±25 |
| 405 | 56±1 | 16±7 | 31±18 | 11±9 |
| 410 | 41±31 | 7±11 | 26±5 | 11±9 |
| 412 | 41±2 | 56±8 | 27±13 | 63±39 |
| 463 | 36±27 | 59±50 | 12±28 | 20±1 |
| 553 | 32±25 | 65±41 | 32±36 | 17±10 |
| 572 | 73±16 | 56±47 | 26±15 | 22±4 |
RRL and the cancer cell-based luciferase assay in HCT116, PC3-MM2, and A549 cells were used to test novobiocin analogues and identify potential Hsp90 inhibitors at a final concentration of 25 μM. The data displayed here represent the mean±standard deviation from two independent experiments (n=2).
Hsp90, heat-shock protein 90; RRL, rabbit reticulocyte lysate.