Diaminopyridine-Based Potent and Selective Mps1 Kinase Inhibitors Binding to an Unusual Flipped-Peptide Conformation

Kusakabe, Ken‐ichi; Ide, Nobuyuki; Daigo, Yataro; Itoh, Takeshi; Higashino, Kenichi; Okano, Yasuhiro; Tadano, G.; Tachibana, Yuki; Sato, Yuji; Inoue, Minako; Wada, Tooru; Iguchi, Motofumi; Kanazawa, Takayuki; Ishioka, Yukichi; Dohi, Keiji; Tagashira, Sachie; Kido, Yasuto; Sakamoto, Shingo; Yasuo, Kazuya; Maeda, Masahiro; Yamamoto, Takahiko; Higaki, Masayo; Endoh, Takeshi; Ueda, Kazuo; Shiota, Takeshi; Murai, Hitoshi; Nakamura, Yusuke

doi:10.1021/ml3000879

Cited by 32 publications

(63 citation statements)

References 25 publications

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“…Cellular studies of these Mps1 inhibitors prove evidence of behaviors expected for a mechanism that induces mitotic checkpoint bypass: a) decrease in the presence of mitotic markers elevated upon taxane-induced mitotic arrest as well as the same markers in asynchronous cultures; b) elevated γ-H2AX; c) shorter mitoses; d) aneuploidy as confirmed by flow cytometry; and e) loss in cell viability and induction of apoptosis. Other Mps1 inhibitors have been reported to modulate the mitotic checkpoint in cellular studies with findings similar to those with PF-7006 and PF-3837 [ 17 , 24 , 26 – 29 , 41 ]. For example, MPI-0479605 is an Mps1 inhibitor shown to induce mitotic checkpoint attenuation and induce aneuploidy in vitro [ 29 ].…”

Section: Discussionsupporting

confidence: 56%

Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility

et al. 2015

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Cell cycle checkpoint intervention is an effective therapeutic strategy for cancer when applied to patients predisposed to respond and the treatment is well-tolerated. A critical cell cycle process that could be targeted is the mitotic checkpoint (spindle assembly checkpoint) which governs the metaphase-to-anaphase transition and insures proper chromosomal segregation. The mitotic checkpoint kinase Mps1 was selected to explore whether enhancement in genomic instability is a viable therapeutic strategy. The basal-a subset of triple-negative breast cancer was chosen as a model system because it has a higher incidence of chromosomal instability and Mps1 expression is up-regulated. Depletion of Mps1 reduces tumor cell viability relative to normal cells. Highly selective, extremely potent Mps1 kinase inhibitors were created to investigate the roles of Mps1 catalytic activity in tumor cells and normal physiology (PF-7006, PF-3837; K i<0.5 nM; cellular IC50 2–6 nM). Treatment of tumor cells in vitro with PF-7006 modulates expected Mps1-dependent biology as demonstrated by molecular and phenotypic measures (reduced pHH3-Ser10 levels, shorter duration of mitosis, micro-nucleation, and apoptosis). Tumor-bearing mice treated with PF-7006 exhibit tumor growth inhibition concomitant with pharmacodynamic modulation of a downstream biomarker (pHH3-Ser10). Unfortunately, efficacy only occurs at drug exposures that cause dose-limiting body weight loss, gastrointestinal toxicities, and neutropenia. Mps1 inhibitor toxicities may be mitigated by inducing G1 cell cycle arrest in Rb1-competent cells with the cyclin-dependent kinase-4/6 inhibitor palbociclib. Using an isogenic cellular model system, PF-7006 is shown to be selectively cytotoxic to Rb1-deficient cells relative to Rb1-competent cells (also a measure of kinase selectivity). Human bone marrow cells pretreated with palbociclib have decreased PF-7006-dependent apoptosis relative to cells without palbociclib pretreatment. Collectively, this study raises a concern that single agent therapies inhibiting Mps1 will not be well-tolerated clinically but may be when combined with a selective CDK4/6 drug.

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Section: Discussionsupporting

confidence: 56%

Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility

et al. 2015

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“…The high-resolution co-crystallography results raised the question of why we were able to generate such a selective inhibitor that lacked a localized conformational change assumed to be a key factor in generating selectivity [16]–[30]. To address if our experimental protocol could detect such localized conformation changes, we performed a control crystallographic study of p38αMAPK in complex with SB239063.…”

Section: Resultsmentioning

confidence: 99%

“…A recently emerging variant of the active site approach focuses on induction of localized conformational changes by inhibitors in order to achieve selectivity [16]–[30]. In the case of p38αMAPK inhibitors, the approach posits that the common theme of exploiting hydrogen (H)-bond interactions with the Met109 amide bond in the phylogenetically conserved kinase hinge region can be enhanced by designing inhibitors that can also engage the contiguous Gly110, thereby yielding highly selective inhibitors that can induce a localized conformational change, termed a glycine flip.…”

Section: Introductionmentioning

confidence: 99%

Development of Novel In Vivo Chemical Probes to Address CNS Protein Kinase Involvement in Synaptic Dysfunction

et al. 2013

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Serine-threonine protein kinases are critical to CNS function, yet there is a dearth of highly selective, CNS-active kinase inhibitors for in vivo investigations. Further, prevailing assumptions raise concerns about whether single kinase inhibitors can show in vivo efficacy for CNS pathologies, and debates over viable approaches to the development of safe and efficacious kinase inhibitors are unsettled. It is critical, therefore, that these scientific challenges be addressed in order to test hypotheses about protein kinases in neuropathology progression and the potential for in vivo modulation of their catalytic activity. Identification of molecular targets whose in vivo modulation can attenuate synaptic dysfunction would provide a foundation for future disease-modifying therapeutic development as well as insight into cellular mechanisms. Clinical and preclinical studies suggest a critical link between synaptic dysfunction in neurodegenerative disorders and the activation of p38αMAPK mediated signaling cascades. Activation in both neurons and glia also offers the unusual potential to generate enhanced responses through targeting a single kinase in two distinct cell types involved in pathology progression. However, target validation has been limited by lack of highly selective inhibitors amenable to in vivo use in the CNS. Therefore, we employed high-resolution co-crystallography and pharmacoinformatics to design and develop a novel synthetic, active site targeted, CNS-active, p38αMAPK inhibitor (MW108). Selectivity was demonstrated by large-scale kinome screens, functional GPCR agonist and antagonist analyses of off-target potential, and evaluation of cellular target engagement. In vitro and in vivo assays demonstrated that MW108 ameliorates beta-amyloid induced synaptic and cognitive dysfunction. A serendipitous discovery during co-crystallographic analyses revised prevailing models about active site targeting of inhibitors, providing insights that will facilitate future kinase inhibitor design. Overall, our studies deliver highly selective in vivo probes appropriate for CNS investigations and demonstrate that modulation of p38αMAPK activity can attenuate synaptic dysfunction.

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“…Using a cell viability assay, we show that the p.I531M-containing AzR3 cell clone was also resistant to the OncoTherapy compound II (ONCOII, patent WO2011016472A1) and NMS-P715 (6) although no resistance was seen to the Shionogi compound 12 (SNG12; ref. 34). The p.S611G-containing AzR1 cells conferred up to 10-fold resistance against the ONCOII and SNG12 inhibitors; however, it showed no resistance against NMS-P715.…”

Section: The Generation Of Az3146-resistant Cell Linesmentioning

confidence: 99%

Naturally Occurring Mutations in the MPS1 Gene Predispose Cells to Kinase Inhibitor Drug Resistance

et al. 2015

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Acquired resistance to therapy is perhaps the greatest challenge to effective clinical management of cancer. With several inhibitors of the mitotic checkpoint kinase MPS1 in preclinical development, we sought to investigate how resistance against these inhibitors may arise so that mitigation or bypass strategies could be addressed as early as possible. Toward this end, we modeled acquired resistance to the MPS1 inhibitors AZ3146, NMS-P715, and CCT251455, identifying five point mutations in the kinase domain of MPS1 that confer resistance against multiple inhibitors. Structural studies showed how the MPS1 mutants conferred resistance by causing steric hindrance to inhibitor binding. Notably, we show that these mutations occur in nontreated cancer cell lines and primary tumor specimens, and that they also preexist in normal lymphoblast and breast tissues. In a parallel piece of work, we also show that the EGFR p.T790M mutation, the most common mutation conferring resistance to the EGFR inhibitor gefitinib, also preexists in cancer cells and normal tissue. Our results therefore suggest that mutations conferring resistance to targeted therapy occur naturally in normal and malignant cells and these mutations do not arise as a result of the increased mutagenic plasticity of cancer cells. Cancer Res; 75(16); 3340-54. Ó2015 AACR.

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Diaminopyridine-Based Potent and Selective Mps1 Kinase Inhibitors Binding to an Unusual Flipped-Peptide Conformation

Cited by 32 publications

References 25 publications

Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility

Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility

Development of Novel In Vivo Chemical Probes to Address CNS Protein Kinase Involvement in Synaptic Dysfunction

Naturally Occurring Mutations in the MPS1 Gene Predispose Cells to Kinase Inhibitor Drug Resistance

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