title: Characterization of kinase inhibitors using different phosphorylation states of colony stimulating factor-1 receptor tyrosine kinase.
abstract: It is known that some kinase inhibitors are sensitive to the phosphorylation state of the kinase, and therefore those compounds can discriminate between a phosphorylated and unphosphorylated protein. In this study, we prepared two colony stimulating factor-1 receptor (CSF-1R) tyrosine kinase proteins: one highly phosphorylated by autophosphorylation and the other dephosphorylated by phosphatase treatment. These kinases were subjected to an activity-based assay to investigate the effect of their phosphorylation state on the potency of several kinase inhibitors. Dasatinib, sorafenib, PD173074 and staurosporine showed similar inhibition against different phosphorylation states of CSF-1R, but pazopanib, sunitinib, GW2580 and imatinib showed more potent inhibition against dephosphorylated CSF-1R. Binding analysis of the inhibitors to the two different phosphorylation forms of CSF-1R, using surface plasmon resonance spectrometry, revealed that staurosporine bound to both forms with similar affinity, but sunitinib bound to the dephosphorylated form with higher affinity. Thus, these observations suggest that sunitinib binds preferentially to the inactive form, preventing the activation of CSF-1R. Screening against different activation states of kinases should be an important approach for prioritizing compounds and should facilitate inhibitor design.
authors: Carna Biosciences, Inc, 1-5-5, Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan. daisuke.kitagawa@carnabio.com
authors: Daisuke
authors: D
authors: Kitagawa
authors: Masaki
authors: M
authors: Gouda
authors: Yasuyuki
authors: Y
authors: Kirii
authors: Naoyuki
authors: N
authors: Sugiyama
authors: Yasushi
authors: Y
authors: Ishihama
authors: Ikuo
authors: I
authors: Fujii
authors: Yugo
authors: Y
authors: Narumi
authors: Kensaku
authors: K
authors: Akita
authors: Koichi
authors: K
authors: Yokota
mesh_terms: D000818:Animals; D001549:Benzamides; D001557:Benzenesulfonates / Q000378:metabolism / Q000494:pharmacology; D001667:Binding, Competitive; D002460:Cell Line; D000069439:Dasatinib; D004305:Dose-Response Relationship, Drug; D006801:Humans; D000068877:Imatinib Mesylate; D007191:Indazoles; D007211:Indoles / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D009536:Niacinamide / Q000031:analogs & derivatives; D010671:Phenylurea Compounds; D010766:Phosphorylation; D010879:Piperazines / Q000378:metabolism / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000378:metabolism* / Q000494:pharmacology*; D011725:Pyridines / Q000378:metabolism / Q000494:pharmacology; D011743:Pyrimidines / Q000378:metabolism / Q000494:pharmacology; D011758:Pyrroles / Q000378:metabolism / Q000494:pharmacology; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D000077157:Sorafenib; D018411:Spodoptera; D019311:Staurosporine / Q000378:metabolism / Q000494:pharmacology; D013449:Sulfonamides / Q000378:metabolism / Q000494:pharmacology; D000077210:Sunitinib; D020349:Surface Plasmon Resonance; D013844:Thiazoles / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection
chemical_list: D001549:Benzamides; D001557:Benzenesulfonates; D007191:Indazoles; D007211:Indoles; C115711:PD 173074; D010671:Phenylurea Compounds; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011743:Pyrimidines; D011758:Pyrroles; D013449:Sulfonamides; D013844:Thiazoles; D009536:Niacinamide; C516667:pazopanib; D000068877:Imatinib Mesylate; D000077157:Sorafenib; D016186:Receptor, Macrophage Colony-Stimulating Factor; D019311:Staurosporine; D000069439:Dasatinib; D000077210:Sunitinib