An allosteric inhibitor of PAK1
NVS-PAK1-1 is an allosteric inhibitor of p21-activated kinase 1 (PAK1), a non-receptor serine/threonine kinase involved in tumorigenesis (Kd = 7 nM).1 It has an IC50 value of 5.2 nM in a PAK1 dephosphorylation assay. NVS-PAK1-1 is selective for PAK1 over PAK2 (Kd = 400 nM) and over a panel of 442 kinases, where it did not inhibit any other kinases by greater than 80%.1,2 It inhibits phosphorylation of MEK Ser289 when used at concentrations ranging from 6 to 20 ?M but does not inhibit proliferation of Su86.86 cells at concentrations lower than 2 ?M.1 See the Structural Genomics Consortium (SGC) website for more information.
1.Karpov, A.S., Amiri, P., Bellamacina, C., et al.Optimization of a dibenzodiazepine hit to a potent and selective allosteric PAK1 inhibitorACS Med. Chem. Lett.6(7)776-781(2015) 2.Semenova, G., and Chernoff, J.Targeting PAK1Biochem. Soc. Trans.45(1)79-88(2017)
Kinase experiment: | Inhibition of PAK1 kinase activity is measured using the Caliper assay. The assay is performed using 384-well microtiter plates. Compounds (NVS-PAK1-1) are tested as 8-point dose responses. The assays are prepared by addition of 50 nL of compound solution in 90% DMSO directly into the empty plate. Subsequently, 4.5 μL of the enzyme solution is added to each well and the resulting solution is pre-incubated at 30°C for 60 min, followed by addition of 4.5 μL of the peptide/ATP-solution. After 60 min incubation at 30°C, reactions are terminated by addition of 16 μL per well of the stop solution. Plates with terminated kinase reactions are transferred to the Caliper LC3000 workstations for reading. Product formation is measured in a microfluidic mobility shift assay. IC50 values are derived from percent inhibition values at different compound concentrations by non-linear regression analysis[1]. |
参考文献: [1]. Karpov AS, et al. Optimization of a Dibenzodiazepine Hit to a Potent and Selective Allosteric PAK1 Inhibitor. ACS Med Chem Lett. 2015 May 22;6(7):776-81. | |
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