Biochemical characterization of GSK1070916, a potent and selective inhibitor of Aurora B and Aurora C kinases with an extremely long residence time1

Abstract

The Aurora kinases AurA, B and C are serine/threonine protein kinases that play essential roles in mitosis and cytokinesis. Among them, AurB is required for maintaining proper chromosome alignment, separation and segregation during mitosis, and regulating a number of critical processes involved in cytokinesis. AurB overexpression has been observed in a variety of cancer cell lines, and inhibition of AurB has been shown to induce tumour regression in mouse xenograft models. In the present study we report the enzymatic characterization of a potent and selective AurB/AurC inhibitor. GSK1070916 is a reversible and ATP-competitive inhibitor of the AurB-INCENP (inner centromere protein) enzyme. It selectively inhibits AurB-INCENP (K(i)*=0.38+/-0.29 nM) and AurC-INCENP (K(i)*=1.5+/-0.4 nM) over AurA-TPX2 (target protein for Xenopus kinesin-like protein 2) (K(i)=490+/-60 nM). Inhibition of AurB-INCENP and AurC-INCENP is time-dependent, with an enzyme-inhibitor dissociation half-life of >480 min and 270+/-28 min respectively. The extremely slow rate of dissociation from the AurB and AurC enzymes distinguishes GSK1070916 from two other Aurora inhibitors in the clinic, AZD1152 and VX-680 (also known as MK-0457).

Biological Characterization of TAK-901, an Investigational, Novel, Multitargeted Aurora B Kinase Inhibitor

“…In contrast, TAK-901 displayed a slow on-rate to inhibit Aurora B/INCENP and once formed, the TAK-901-bound Aurora B/INCENP complex dissociated slowly with a half-life of 920 minutes. This favorable biochemical property of extremely long residence time on Aurora B/INCENP was also reported recently for GSK1070916, an Aurora B/Cselective kinase inhibitor (26). Although the biochemical methods used were different, the TAK-901 dissociation rate seems of comparable magnitude with GSK1070916, whereas their kinetic analysis indicated that Aurora kinase inhibitors AZD1152 and VX-680/MK-0457 exhibit much faster dissociation rates (26).…”

“…This favorable biochemical property of extremely long residence time on Aurora B/INCENP was also reported recently for GSK1070916, an Aurora B/Cselective kinase inhibitor (26). Although the biochemical methods used were different, the TAK-901 dissociation rate seems of comparable magnitude with GSK1070916, whereas their kinetic analysis indicated that Aurora kinase inhibitors AZD1152 and VX-680/MK-0457 exhibit much faster dissociation rates (26). Therefore, the Aurora B tight-binding property may be less common among reported Aurora B kinase inhibitors.…”

“…However, many pan-Aurora kinase inhibitors generate a similar polyploid cellular phenotype, which is a hallmark of Aurora B inactivation (11,14). More than 10 small-molecule inhibitors have entered early clinical development, including selective inhibitors of Aurora A (23)(24)(25), B, or C (26)(27)(28)(29) and panAurora kinase inhibitors (30)(31)(32)(33)(34)(35).…”

Aurora B: A New Prognostic Marker and Therapeutic Target in Cancer

“…Selective inhibitors of Aurora B have been developed: AZD1152 [136] (highly potent and selective inhibitor of Aurora B), and GSK1070916 (a potent and selective inhibitor of Aurora B and Aurora C kinases) [137]. The general chemical structures of selective Aurora B inhibitors are shown in (Fig.…”

Quick Evaluation of Kinase Inhibitors by Surface Plasmon Resonance Using Single-Site Specifically Biotinylated Kinases

“…One clue to the proper correlation between in vitro and in vivo drug activity is the dissociation rate, which may better describe cellular efficacy than K D and IC 50 among kinase inhibitors and other type of enzyme inhibitors such as proteases. 3,15 A good example of this is given in the study of the AIDS drug saquinavir. Although saquinavir is an effective HIV protease inhibitor in treating AIDS, resistant mutants arise through the treatment.…”