Discovery of the First Potent and Selective Inhibitor of Centromere-Associated Protein E: GSK923295

Qian, Xiangping; McDonald, Andrew; Zhou, Han-Jie; Adams, Nicholas D.; Parrish, Cynthia A.; Duffy, Kevin J.; Fitch, Duke M.; Tedesco, Rosanna; Ashcraft, Luke; Yao, Bing; Jiang, Hong; Huang, Jennifer; Marin, Melchor V.; Aroyan, Carrie E.; Wang, Jianchao; Ahmed, Seyed; Burgess, Joelle L.; Chaudhari, Amita M.; Donatelli, Carla A.; Darcy, Michael G.; Ridgers, Lance H.; Newlander, Kenneth A.; Schmidt, Stanley J.; Chai, Deping; Colón, Mariela; Zimmerman, Michael N.; Lad, Latesh; Sakowicz, Roman; Schauer, Stephen; Belmont, Lisa D.; Baliga, Ramesh; Pierce, Daniel W.; Finer, Jeffrey T.; Wang, Zhengping; Morgan, Bradley; Morgans, David J.; Auger, Kurt R.; Sung, Chiu‐Mei; Carson, Jeff D.; Luo, Liang; Hugger, Erin; Copeland, Robert A.; Sutton, David; Elliott, John D.; Jackson, Jeffrey R.; Wood, Kenneth W.; Dhanak, Dashyant; Bergnes, Gustave; Knight, Steven D.

doi:10.1021/ml900018m

Cited by 61 publications

(56 citation statements)

References 29 publications

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“…Such ATP-competitive behavior had been observed during the optimization of this chemical series of CENP-E inhibitors (16). Remarkably, chemical modifications as small as a one carbon extension of a sidechain were sufficient to alter the steady-state mechanism of inhibition from ATP-uncompetitive to ATP-competitive behavior (16). Such minor changes seemed (Table 1, Table S1, and Fig.…”

Section: Resultsmentioning

confidence: 66%

“…Medicinal chemistry optimization of initial inhibitors gave rise to a series of high affinity, stereoselective, specific and drug-like inhibitors of CENP-E with potent on-target activity against human cancer cells, culminating in the identification of GSK923295 and related compounds ( Fig. 1 and Table S1) (16).…”

Section: Resultsmentioning

confidence: 99%

“…Although we have not excluded the possibility that GSK923295 and related compounds might affect the function of other undetermined mitotic proteins, the general correlation of biochemical and cellular activity across this structurally related chemical series, including stereoselective inhibition of both CENP-E ATPase and cell growth, supports CENP-E as the biologically relevant target (16). These observations, coupled with the selectivity of GSK923295 for CENP-E among seven diverse kinesins tested, the similarity of phenotype to that produced by knockdown of CENP-E protein, and interaction with a structurally unique allosteric binding site, support our conclusion that CENP-E is the target responsible for GSK923295 antimitotic and antitumor activity.…”

Section: +mentioning

confidence: 95%

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Antitumor activity of an allosteric inhibitor of centromere-associated protein-E

Wood

Lad²,

Luo³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Centromere-associated protein-E (CENP-E) is a kinetochore-associated mitotic kinesin that is thought to function as the key receptor responsible for mitotic checkpoint signal transduction after interaction with spindle microtubules. We have identified GSK923295, an allosteric inhibitor of CENP-E kinesin motor ATPase activity, and mapped the inhibitor binding site to a region similar to that bound by loop-5 inhibitors of the kinesin KSP/Eg5. Unlike these KSP inhibitors, which block release of ADP and destabilize motor-microtubule interaction, GSK923295 inhibited release of inorganic phosphate and stabilized CENP-E motor domain interaction with microtubules. Inhibition of CENP-E motor activity in cultured cells and tumor xenografts caused failure of metaphase chromosome alignment and induced mitotic arrest, indicating that tight binding of CENP-E to microtubules is insufficient to satisfy the mitotic checkpoint. Consistent with genetic studies in mice suggesting that decreased CENP-E function can have a tumor-suppressive effect, inhibition of CENP-E induced tumor cell apoptosis and tumor regression.entromere-associated protein-E (CENP-E; kinesin-7) is a kinetochore-associated kinesin motor protein with an essential and exclusive role in metaphase chromosome alignment and satisfaction of the mitotic checkpoint (1). CENP-E is a likely candidate to integrate the mechanics of kinetochore-microtubule interaction with the mitotic checkpoint signaling machinery responsible for restraining cell-cycle progression into anaphase. CENP-E is a large dimeric protein consisting of an N-terminal kinesin motor domain tethered to a globular C-terminal domain through an extended coiled-coil rod domain (2, 3). The C-terminal, noncatalytic region of CENP-E is not only sufficient to specify localization to kinetochores, but it also mediates interaction of CENP-E with the serine/threonine kinase BubR1, a key effector of mitotic checkpoint signaling that forms complexes with the checkpoint proteins Cdc20, Bub3, and Mad2 to inhibit the ubiquitin ligase activity of the anaphase promoting complex APC/C CDC20 (4-7). The combined interaction of CENP-E with microtubules and a key regulator of APC/C CDC20 has led to the hypothesis that CENP-E functions as the key kinetochore microtubule receptor responsible for silencing mitotic checkpoint signal transduction after capture of spindle microtubules. This hypothesis was further strengthened by the finding that CENP-E could stimulate the kinase activity of BubR1 in a microtubule-sensitive manner (8, 9). In vitro, the addition of CENP-E to BubR1 resulted in a stimulation of BubR1 kinase activity. The addition of microtubules suppressed this stimulatory activity, an effect thought to be mediated by the CENP-E kinesin motor domain. Although the importance of CENP-E interaction with BubR1 and the role of BubR1-mediated phosphorylation in mitotic checkpoint function remain unclear, CENP-E remains a prominent candidate to play a key role in mitotic checkpoint signal transduction.Depletion of CENP-E from ...

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Section: Resultsmentioning

confidence: 66%

Section: Resultsmentioning

confidence: 99%

Section: +mentioning

confidence: 95%

See 1 more Smart Citation

Antitumor activity of an allosteric inhibitor of centromere-associated protein-E

Wood

Lad²,

Luo³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

216

219

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“…CENP-E is expressed predominantly in mitosis (and G2) [290] and plays an important role in peripheral chromosome congression [293,295,300], thereby representing an attractive target for cancer therapeutics. GSK923295 is an allosteric inhibitor of CENP-E that blocks its microtubule stimulated ATPase activity and stabilizes the interaction between the motor domain and microtubules [449,450]. GSK923295 has demonstrated both in vitro and in vivo antitumor activity against various malignancies [449,451,452,453,454,455].…”

Section: Targeting Chromosome Congression For Cancer Therapymentioning

confidence: 99%

Mechanisms of Chromosome Congression during Mitosis

Maiato

Gomes

Sousa

et al. 2017

Biology

174

176

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Chromosome congression during prometaphase culminates with the establishment of a metaphase plate, a hallmark of mitosis in metazoans. Classical views resulting from more than 100 years of research on this topic have attempted to explain chromosome congression based on the balance between opposing pulling and/or pushing forces that reach an equilibrium near the spindle equator. However, in mammalian cells, chromosome bi-orientation and force balance at kinetochores are not required for chromosome congression, whereas the mechanisms of chromosome congression are not necessarily involved in the maintenance of chromosome alignment after congression. Thus, chromosome congression and maintenance of alignment are determined by different principles. Moreover, it is now clear that not all chromosomes use the same mechanism for congressing to the spindle equator. Those chromosomes that are favorably positioned between both poles when the nuclear envelope breaks down use the so-called “direct congression” pathway in which chromosomes align after bi-orientation and the establishment of end-on kinetochore-microtubule attachments. This favors the balanced action of kinetochore pulling forces and polar ejection forces along chromosome arms that drive chromosome oscillatory movements during and after congression. The other pathway, which we call “peripheral congression”, is independent of end-on kinetochore microtubule-attachments and relies on the dominant and coordinated action of the kinetochore motors Dynein and Centromere Protein E (CENP-E) that mediate the lateral transport of peripheral chromosomes along microtubules, first towards the poles and subsequently towards the equator. How the opposite polarities of kinetochore motors are regulated in space and time to drive congression of peripheral chromosomes only now starts to be understood. This appears to be regulated by position-dependent phosphorylation of both Dynein and CENP-E and by spindle microtubule diversity by means of tubulin post-translational modifications. This so-called “tubulin code” might work as a navigation system that selectively guides kinetochore motors with opposite polarities along specific spindle microtubule populations, ultimately leading to the congression of peripheral chromosomes. We propose an integrated model of chromosome congression in mammalian cells that depends essentially on the following parameters: (1) chromosome position relative to the spindle poles after nuclear envelope breakdown; (2) establishment of stable end-on kinetochore-microtubule attachments and bi-orientation; (3) coordination between kinetochore- and arm-associated motors; and (4) spatial signatures associated with post-translational modifications of specific spindle microtubule populations. The physiological consequences of abnormal chromosome congression, as well as the therapeutic potential of inhibiting chromosome congression are also discussed.

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“…These findings also raise the intriguing possibility that native cellular ligands may exist that bind at this site and regulate kinesins. Another notable point is that this class of CENP-E inhibitors can switch between ATPuncompetitive and ATP-competitive inhibition with chemical modifications as small as a single carbon extension (16).…”

mentioning

confidence: 99%