BAY1125976

2D structure
Target AKT1  AKT2  AKT3 
Targeted domain Kinase domain (allosteric pocket)
Mode of action Covalent inhibitor (type 4, non-ATP competitive)
Control BAY-940
Orthogonal probe Borussertib 
Recommended cellular usage concentration ≤ 1 µM
In vivo use Yes
Donated by Bayer


Probe criteria


Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) Surpasses criterion: TR-FRET with full-length protein: AKT1 (IC50 = 5.2 nM at 10 µM ATP and 44 nM at 2 mM ATP); AKT2 (IC50 = 18 nM at 10 µM ATP and 36 nM at 2 mM ATP)
SPR: active AKT1 (Kd = 2.7 nM); inactive AKT1 (Kd = 1.3 nM)
Truncated AKTs lacking the PH domain (TR-FRET): ΔPH-AKT1 (IC50 >20 µM), ΔPH-AKT2 (IC50 >20 µM)
AKT3: IC50 = 427 nM at 10 µM ATP (TR-FRET) (Bioduro)
Selectivity within target family: > 30-fold Millipore KinaseProfilerTM panel (230 kinases at 10 µM): FLT1, FLT3, FLT4 and MERTK > 50 % inhibition
DiscoverX (468 kinases): Kd [nM] (FLT(D835Y) 210), CLK1 (310) and MKNK2 (330)
Selectivity outside target family Ricerca Lead Profiler screen: No effect < 10 µM
Clean PDSP scan with one off-target: CHRM1 (Ki = 1454.31 nM)
On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 Surpasses criterion: NanoBRET assay (n=3; SGC Frankfurt): AKT1 (EC50 =15 ± 4.3 nM), AKT2 (EC50 = 30 ± 4.7 nM), AKT3 (EC50 = 200 ± 72 nM)
Mechanistic cell assay:
  1. Reduction of the basal levels of AKT phosphorylation in KPL-4 cells at T308 (phosphorylated exclusively by PI3K/PDK1): IC50 = 0.9 nM
  2. Reduction of the basal levels of AKT phosphorylation in KPL-4 cells at S473 (phosphorylated by PI3K/PDK1 and mTORC2): IC50 = 1.1 nM
  3. Blocking of the activation of the downstream signaling molecule 4EBP1 by phosphorylation at mTOR substrate T70: IC50 = 35 nM
Complete inhibition of phosphorylation by PDK1 of inactive AKT
Inhibition of PRAS40 phosphorylation at T246 (direct target of AKT1 activity) in LAPC-4 cells: IC50 ~141 nM
Inhibition of activation of AKT in cell lines carrying the AKT-activating mutation E17K:
  1. KU-19–19 bladder cancer cells: Inhibition of activation by phosphorylation: IC50 = 35 nM (AKT1-S473) and IC50 = 100 nM (4EBPI-T70)
  2. Prostate cancer cell line LAPC-4: Inhibition of phosphorylation: IC50 = 0.8 nM (AKT1-S473), IC50 = 5.6 nM (T308), IC50 = 35 nM (4EBPI-T70)
Control compound (100 times less potent than the probe) BAY-940: NanoBRET assay (n=3; SGC Frankfurt): AKT1 (EC50 = 1300 ± 29 nM), AKT2 (EC50 = 4200 ± 250 nM, AKT3 (EC50 > 50 µM)
Biochemical assay: AKT1 (IC50 = 3.74 µM) and AKT2 (IC50 = 3.84 µM)