tak-632 and dabrafenib

The BRAF inhibitors dabrafenib and vemurafenib induce remarkable clinical responses in patients with BRAF-mutated melanomas. However, adverse events, including the emergence of secondary tumors and drug resistance, have been reported. Studies have revealed that undesirable RAF dimerization induced by inhibitors promotes these adverse effects. Here, we developed highly sensitive biosensors of RAF dimerization in cells utilizing the split enhanced click beetle luciferase (Emerald Luc, ELuc) complementation technique. We demonstrated that our biosensor system works effectively for high-throughput screens in the microplate format. A comprehensive analysis of commercially available RAF inhibitors performed using this assay system revealed that the inhibitors exhibit various potencies in inducing the dimerization of RAF isoforms, and their dimerization potencies do not always correlate with the RAF enzyme inhibition. This sensitive assay system will become a powerful tool to discover next-generation BRAF inhibitors with safer profiles.

Topics: Benzothiazoles; Biosensing Techniques; Cell Line, Tumor; Dimerization; Heterocyclic Compounds, 2-Ring; Humans; Imidazoles; Melanoma; Nitriles; Oximes; Phenylurea Compounds; Protein Kinase Inhibitors; Protein Multimerization; Proto-Oncogene Proteins B-raf; Pyrimidines; Sulfonamides; Vemurafenib