Table 2.
Biochemical and biological activities of compound 2 and its derivatives
| Compound number |
KD* and Ki* (μmol/L) |
pAKT inhibition† (IC50, μmol/L) |
Apoptosis‡ at 20 μmol/L (%) |
Cell survival§ (IC50, μmol/L) |
|---|---|---|---|---|
| 2 |
KD = 0.45 ± 0.1 Ki > 50.0 |
20/25 | 24.3 ± 3.2/25.7 ± 2.6 | NI/NI |
| 24 |
KD = 19.6 ± 4.9 Ki = 21.8 ± 1.8 |
10/15 | 28.7 ± 0.3/20.0 ± 1.5 | 127/90 |
| 25 |
KD = NB Ki > 50 |
>50/>50 | 6.8 ± 0.9/10.3 ± 2.1 | NI/NI |
| 26 |
KD = NB Ki > 50 |
>50/>50 | 11.4 ± 0.5/18.7 ± 3.1 | NI/NI |
| 27 |
KD = 40.8 ± 2.5 Ki = 2.4 ± 0.6 |
6.3 ± 0.9/10 | 40.0 ± 2.9/31.3 ± 1.6 | 65/30 |
NOTE: All biological tests were made in Panc-1 (numbers on the left) and MiaPaCa-2 (number on the right) pancreatic cell lines.
Abbreviations: NI, not inhibitory; NB, not binding.
*
KD and Ki (μmol/L) were determined using purified AKT PH domain and SPR spectroscopy (Biacore 2000). The Ki for PtdIns(3,4,5)trisphosphate was 0.26 μmol/L.
†
Inhibition of AKT was measured by Western blots using specific antibodies against phospho-Ser473-AKT.
‡
Percentage of apoptosis was obtained by a morphologic assay at 20 μmol/L.
§
Cell survival was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.