Bioluminescent Kinase Profiling Systems For Characterizing Small Molecule Kinase Inhibitors
Poster Aug 07, 2017
Hicham Zegzouti, Jacquelyn Hennek, Tracy Worzella, Monse Contreras, Cristopher Cowan and Said Goueli
In order to profile compounds against a broad panel of kinases, in-house profiling requires rigorous kinase assay development. Most importantly, it requires an optimization for each kinase in the panel, which can be costly and time consuming. On the other hand, outsourcing kinase profiling is fraught with obstacles such as requirements of agreements, long timelines and lack of control over the whole process. Thus, a profiling system with simple and rapid in-house implementation would obviate such logistical inconveniences and concerns.
We created new kinase profiling systems based on the luminescent ADP-Glo™ kinase assay platform. The kinase profiling systems have the following features and advantages:
• Set of kinases organized by kinase families, presented in easy to use multi-well strips, and standardized for optimal kinase activity.
• The strip system provides flexible kinase inhibitor profiling, as each strip can be used to profile compounds at a single dose or used for a dose response against 8 kinases at once.
• Easily automated with fast and simple reaction assembly.
The data generated with this novel set-up are concordant with published inhibitor potency profiles produced by radioactivity assays. Using this technology we created profiles for 16 small molecule compounds that are approved for different cancers and inflammatory diseases. Medicinal chemists and chemical biologists can easily adopt this novel approach for regular in-house kinase inhibitor profiling and gain more control over the data for fast progression into developing lead compounds.
Primary Human Hepatocyte (PHH) culture provides the closest in vitro model to human liver that can produce a metabolic profile of a given drug very similar to that found in vivo. Recently we have developed an easy-to-assemble user-friendly in vitro Primary Human Hepatocyte (PHH) 3D-spheroid modelREAD MORE