Potency data for probe BAY1125976 and control BAY-940
In vitro potency
| Target name | Target information | Species | Assay description | BAY1125976 | BAY-940 | Publication of assay conditions |
|---|---|---|---|---|---|---|
| AKT1 | Invitrogen; full-length | Homo sapiens | TR-FRET (AKT1, 10 uM ATP): TR-FRET-based kinase activity assay with the biotinylated peptide biotin-Ahx-KKLNRTLSFAEPG-NH2 as substrate. Assay with 10 µM ATP | IC50 = 5.2 nM | PMID: 27699769 | |
| AKT1 | Invitrogen; full-length | Homo sapiens | TR-FRET (AKT1, 2 mM ATP): TR-FRET-based kinase activity assay with the biotinylated peptide biotin-Ahx-KKLNRTLSFAEPG-NH2 as substrate. Assay with 2 mM ATP | IC50 = 44 nM | PMID: 27699769 | |
| AKT2 | Invitrogen; full-length | Homo sapiens | TR-FRET (AKT2, 10 uM ATP): TR-FRET-based kinase activity assay with the biotinylated peptide biotin-Ahx-KKLNRTLSFAEPG-NH2 as substrate. Assay with 10 µM ATP | IC50 = 18 nM | PMID: 27699769 | |
| AKT2 | Invitrogen; full-length | Homo sapiens | TR-FRET (AKT2, 2 mM ATP): TR-FRET-based kinase activity assay with the biotinylated peptide biotin-Ahx-KKLNRTLSFAEPG-NH2 as substrate. Assay with 2 mM ATP | IC50 = 36 nM | PMID: 27699769 | |
| AKT1 | active protein; full-length | Homo sapiens | SPR (active AKT1) | Kd = 2.7 nM | PMID: 27699769 | |
| AKT1 | inactive protein; full-length | Homo sapiens | SPR (inactive AKT1) | Kd = 1.3 nM | PMID: 27699769 | |
| AKT1 | AKT1 without PH domain; truncated | Homo sapiens | TR-FRET (delta PH domain AKT1) | IC50 > 20 µM | PMID: 27699769 | |
| AKT2 | AKT2 without PH domain; truncated | Homo sapiens | TR-FRET (delta PH domain AKT2) | IC50 > 20 µM | PMID: 27699769 | |
| AKT3 | full-length | Homo sapiens | TR-FRET (AKT3, 10 µM ATP): Performed at Bioduro | IC50 = 427 nM | PMID: 27699769 | |
| AKT1 | Homo sapiens | Biochemical assay (AKT1) | IC50 = 3.74 µM | |||
| AKT2 | Homo sapiens | Biochemical assay (AKT2) | IC50 = 3.84 µM |
In cell potency
| Target name | Target information | Species | Assay description | BAY1125976 | BAY-940 | Publication of assay conditions |
|---|---|---|---|---|---|---|
| KPL-4 cells (breast cancer), AKT1 (WT) | Homo sapiens | Mechanistic cell assay: Reduction of the basal levels of AKT phosphorylation in KPL-4 cells at T308: Phosphorylated exclusively by PI3K/PDK1/ Phsphorylation level determined by MSD MultiArray System assay kit | IC50 = 0.9 nM | PMID: 27699769 | ||
| KPL-4 cells (breast cancer), AKT1 (WT) | Homo sapiens | Mechanistic cell assay: Reduction of the basal levels of AKT phosphorylation in KPL-4 cells at S473: Phosphorylated by PI3K/PDK1 and mTORC2/ Phsphorylation level determined by MSD MultiArray System assay kit | IC50 = 1.1 nM | PMID: 27699769 | ||
| KPL-4 cells (breast cancer), AKT1 (WT) | Homo sapiens | Mechanistic cell assay: Blocking of the activation of the downstream signaling molecule 4EBP1 by phosphorylation at mTOR substrate T70: Phsphorylation level determined by AlphaScreen (SureFire) assay kit | IC50 = 35 nM | PMID: 27699769 | ||
| KU-19–19 bladder cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: Inhibition of activation by phosphorylation (AKT1-S473) | IC50 = 35 nM | PMID: 27699769 | ||
| KU-19–19 bladder cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: Inhibition of activation by phosphorylation (4EBPI-T70) | IC50 = 100 nM | PMID: 27699769 | ||
| LAPC-4 prostate cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: Inhibition of phosphorylation (AKT1-S473) | IC50 = 0.8 nM | PMID: 27699769 | ||
| LAPC-4 prostate cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: Inhibition of phosphorylation (T308) | IC50 = 5.6 nM | PMID: 27699769 | ||
| LAPC-4 prostate cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: Inhibition of phosphorylation (4EBPI-T70) | IC50 = 35 nM | PMID: 27699769 | ||
| LAPC-4 prostate cancer cells (carrying the AKT-activating mutation E17K) | Homo sapiens | Mechanistic cell assay: inhibition of PRAS40 phosphorylation at T246: PRAS40 phosphorylation at T246 is a direct target of AKT1 activity. | IC50 = 141 nM | PMID: 27699769 | ||
| AKT1 | Homo sapiens | NanoBRET (AKT1): SGC Frankfurt | EC50 = 15 nM ± 4.300 nM (n=3) | EC50 = 1300 nM ± 29.000 nM (n=3) | ||
| AKT2 | Homo sapiens | NanoBRET (AKT2): SGC Frankfurt | EC50 = 30 nM ± 4.700 nM (n=3) | EC50 = 4200 nM ± 250.000 nM (n=3) | ||
| AKT3 | Homo sapiens | NanoBRET (AKT3): SGC Frankfurt | EC50 = 200 nM ± 72.000 nM (n=3) | EC50 > 50 µM (n=3) |