Borussertib
| Target | AKT1 AKT2 AKT3 |
| Targeted domain | Kinase domain (allosteric pocket) |
| Mode of action | Covalent inhibtor (type 4, non-ATP competitive) |
| Control | RL2578 |
| Orthogonal probe | BAY1125976 |
| Recommended cellular usage concentration | 1 µM |
| In vivo use | Yes |
| Donated by | TU Dortmund |
Probe criteria
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: NanoBRET assay (n=3; SGC Frankfurt): EC50 = 26 ± 3.8 nM (AKT1), 250 ± 170 nM (AKT2), 29 ± 11 nM (AKT3)CellTiter-Glo luminescent cell viability assay (CTG): EC50 = 5 ± 1 nM (ZR-75-1, breast), 50 ± 15 nM (T-47D, breast), 190 ± 90 nM (AN3-CA, endometrium), 370 ± 50 (BT-474, breast), 280 ± 90 (MCF-7, breast) |
| Control compound (100 times less potent than the probe) | RL2578: IC50 = 9.2 µM (HTRF), >1000-fold less potent in vitro compared to BorussertibCTG assay: EC50 = 4.2 µM (ZR-75-1), 18 µM (AN3-CA); NanoBRET (n=3; SGC Frankfurt): EC50 = 3.5 ± 1.5 µM (AKT1), 4.8 ± 0.9 µM (AKT2), 9.3 ± 0.66 µM (AKT3) |
| Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) | Surpasses criterion: Homogeneous Time Resolved Fluorescence (HTRF): AKT1: IC50 = 0.8 ± 0.3 nM; Ki = 2.2 ± 0.3 nM; AKT2: IC50 = 59 ± 16 nM; AKT3: IC50 = 650 ± 170 nMKinetic profile for AKT1: kinact (0.111 ± 0.020 1/min); kinact/Ki (0.853 ± 0.038 1/(µM * s)) |
| Selectivity within target family: > 30-fold | SelectScreen® Kinase Profiling (100) at 1 µM: clean |
| Selectivity outside target family | Closest off-targets in the PDSP scan (44 targets) at 10 µM are GABA/PBR (Ki = 57.32 nM), HRH2 (Ki = 949.61 nM), ADRA1A (Ki = 2655.83 nM) and SLC6A3 (Ki = 3655.95 nM). |