Displacement assay for the detection of stabilizers of inactive kinase conformations.
Article Details
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Kluter S, Grutter C, Naqvi T, Rabiller M, Simard JR, Pawar V, Getlik M, Rauh D
Displacement assay for the detection of stabilizers of inactive kinase conformations.
J Med Chem. 2010 Jan 14;53(1):357-67. doi: 10.1021/jm901297e.
- PubMed ID
- 19928858 [ View in PubMed]
- Abstract
Targeting protein kinases with small molecules outside the highly conserved ATP pocket to stabilize inactive kinase conformations is becoming a more desirable approach in kinase inhibitor research, since these molecules have advanced pharmacological properties compared to compounds exclusively targeting the ATP pocket. Traditional screening approaches for kinase inhibitors are often based on enzyme activity, but they may miss inhibitors that stabilize inactive kinase conformations by enriching the active state of the kinase. Here we present the development of a kinase binding assay employing a pyrazolourea type III inhibitor and enzyme fragment complementation (EFC) technology that is suitable to screen stabilizers of enzymatically inactive kinases. To validate this assay system, we report the binding characteristics of a series of kinase inhibitors to inactive p38alpha and JNK2. Additionally, we present protein X-ray crystallography studies to examine the binding modes of potent quinoline-based DFG-out binders in p38alpha.
DrugBank Data that Cites this Article
- Binding Properties
Drug Target Property Measurement pH Temperature (°C) 1-(5-Tert-Butyl-2-Methyl-2h-Pyrazol-3-Yl)-3-(4-Chloro-Phenyl)-Urea Mitogen-activated protein kinase 14 IC 50 (nM) 21000 N/A N/A Details 1-(5-Tert-Butyl-2-Methyl-2h-Pyrazol-3-Yl)-3-(4-Chloro-Phenyl)-Urea Mitogen-activated protein kinase 14 IC 50 (nM) 16800 N/A N/A Details 1-[1-(3-aminophenyl)-3-tert-butyl-1H-pyrazol-5-yl]-3-naphthalen-1-ylurea Mitogen-activated protein kinase 14 Kd (nM) 12 N/A N/A Details 1-[1-(3-aminophenyl)-3-tert-butyl-1H-pyrazol-5-yl]-3-naphthalen-1-ylurea Mitogen-activated protein kinase 14 IC 50 (nM) 90 N/A N/A Details 1-[1-(3-aminophenyl)-3-tert-butyl-1H-pyrazol-5-yl]-3-naphthalen-1-ylurea Mitogen-activated protein kinase 14 IC 50 (nM) 170 N/A N/A Details Doramapimod Mitogen-activated protein kinase 14 IC 50 (nM) 250 N/A N/A Details Doramapimod Mitogen-activated protein kinase 14 IC 50 (nM) 220 N/A N/A Details