High-Throughput Kinase Profiling: A More Efficient Approach toward the Discovery of New Kinase Inhibitors

Miduturu, Chandrasekhar V.; Deng, Xianming; Kwiatkowski, Nicholas; Yang, Wannian; Brault, Laurent; Filippakopoulos, P.; Chung, Eunah; Yang, Qingkai; Schwaller, Juerg; Knapp, Stefan; King, Randall W.; Lee, Jiing-Dwan; Herrgård, Sanna; Zarrinkar, Patrick P.; Gray, Nathanael S.

doi:10.1016/j.chembiol.2011.05.010

Cited by 108 publications

(104 citation statements)

References 55 publications

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“…This suggests that that the kinase activity of HIPK2 is important for the effect, and that specific targeting of HIPK2, which has been recently attempted and lead compounds identified via high-throughput kinase profiling (Miduturu et al 2011), could have therapeutic value for prostate cancer patients where HIPK2 is activated. In fact, sorafenib has shown moderate activity as a second-line treatment for CRPC (Aragon-Ching et al 2009), and our findings suggest that stratifying patients for HIPK2 up-regulation could enhance sorafenib efficacy.…”

Section: Discussionmentioning

confidence: 99%

A genome-wide RNA interference screen identifies new regulators of androgen receptor function in prostate cancer cells

Imberg-Kazdan

Greenfield

et al. 2013

Genome Res.

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The androgen receptor (AR) is a mediator of both androgen-dependent and castration-resistant prostate cancers. Identification of cellular factors affecting AR transcriptional activity could in principle yield new targets that reduce AR activity and combat prostate cancer, yet a comprehensive analysis of the genes required for AR-dependent transcriptional activity has not been determined. Using an unbiased genetic approach that takes advantage of the evolutionary conservation of AR signaling, we have conducted a genome-wide RNAi screen in Drosophila cells for genes required for AR transcriptional activity and applied the results to human prostate cancer cells. We identified 45 AR-regulators, which include known pathway components and genes with functions not previously linked to AR regulation, such as HIPK2 (a protein kinase) and MED19 (a subunit of the Mediator complex). Depletion of HIPK2 and MED19 in human prostate cancer cells decreased AR target gene expression and, importantly, reduced the proliferation of androgen-dependent and castration-resistant prostate cancer cells. We also systematically analyzed additional Mediator subunits and uncovered a small subset of Mediator subunits that interpret AR signaling and affect AR-dependent transcription and prostate cancer cell proliferation. Importantly, targeting of HIPK2 by an FDA-approved kinase inhibitor phenocopied the effect of depletion by RNAi and reduced the growth of AR-positive, but not AR-negative, treatment-resistant prostate cancer cells. Thus, our screen has yielded new AR regulators including drugable targets that reduce the proliferation of castration-resistant prostate cancer cells.

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Section: Discussionmentioning

confidence: 99%

A genome-wide RNA interference screen identifies new regulators of androgen receptor function in prostate cancer cells

Imberg-Kazdan

Greenfield

et al. 2013

Genome Res.

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“…Both inhibitors show specific binding toward PIM1 and PIM2 (K00135 IC 50 values: PIM1, 0.12 mmol/L; PIM2, 1.8 mmol/L; K00486 IC 50 values: PIM1, 0.04 mmol/L; PIM2, 2.5 mmol/L) in in vitro kinase assays. The small-molecule inhibitor A47 is based on another structural background, a furan thiazolidin (39). Besides good efficacy on PIM1 and PIM2, this inhibitor also blocks PIM3 kinase activity (IC 50 values: PIM1, 3 mmol/L; PIM2, 0.8 mmol/L; PIM3, 2.6 mmol/L).…”

Section: Small-molecule Inhibitors Against Pim Kinases and Cxcr4 Antamentioning

confidence: 99%

“…K00135 and K00486, two imidazo[1,2-b]pyridazines, show low IC 50 values for inhibition of PIM1 and PIM2 kinase activity, and have shown proapoptotic effects in AML cells (37,42). The third PIM kinase inhibitor, A47, is based on another chemical structure, a furan thiazolidin, and inhibits all three PIM kinases in low micromolar concentrations including PIM3 (39). Treatment of CLL cells (n ¼ 16) with PIM inhibitors for 48 hours showed a dose-dependent increase in apoptosis (Fig.…”

Section: Pim Kinase Inhibition With Three Different Pim Inhibitors Inmentioning

confidence: 99%

“…To verify the importance of PIM kinase activity for the survival of CLL cells, we investigated the potential of three different small-molecule PIM kinase inhibitors on apoptosis induction in CLL cells (34,36,39). K00135 and K00486, two imidazo[1,2-b]pyridazines, show low IC 50 values for inhibition of PIM1 and PIM2 kinase activity, and have shown proapoptotic effects in AML cells (37,42).…”

Section: Pim Kinase Inhibition With Three Different Pim Inhibitors Inmentioning

confidence: 99%

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PIM Kinases Are Essential for Chronic Lymphocytic Leukemia Cell Survival (PIM2/3) and CXCR4-Mediated Microenvironmental Interactions (PIM1)

Decker

Finter

Forde

et al. 2014

Molecular Cancer Therapeutics

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Overexpression of the CXCR4 receptor is a hallmark of chronic lymphocytic leukemia (CLL) and is important for CLL cell survival, migration, and interaction with their protective microenvironment. In acute myelogenous leukemia (AML), PIM1 was shown to regulate the surface expression of the CXCR4 receptor. Here, we show that PIM (proviral integration site for Moloney murine leukemia virus) kinases 1-3 are overexpressed and that the CXCR4 receptor is hyperphosphorylated on Ser339 in CLL compared with normal lymphocytes. Furthermore, CXCR4 phosphorylation correlates with PIM1 protein expression and PIM1 transcript levels in CLL. PIM kinase inhibition with three different PIM kinase inhibitors induced apoptosis in CLL cells independent of the presence of protective stromal cells. In addition, PIM inhibition caused dephosphorylation of the CXCR4 receptor on Ser339, resulting in enhanced ligand-dependent CXCR4 internalization and reduced re-externalization after withdrawal of CXCL12. Furthermore, PIM inhibition in CLL cells blocked CXCR4 functions, such as migration toward CXCL12-or CXCL12-induced extracellular signal-regulated kinase (ERK) phosphorylation. In concordance, pretreatment of CLL cells with PIM kinase inhibitors strongly reduced homing of CLL cells toward the bone marrow and the spleen of Rag2

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“…To assess the kinase selectivity of FIIN-2 and FIIN-3 broadly, they were profiled against a diverse panel of 456 kinases [DiscoveRX, KinomeScan (53,54)] using an in vitro ATP-site competition binding assay at a concentration of 1.0 μM. Compared with FIIN-1, both compounds displayed strong binding to FGFRs and exhibited good overall kinase selectivity (SI Appendix, Table S1).…”

Section: Significancementioning

confidence: 99%

Development of covalent inhibitors that can overcome resistance to first-generation FGFR kinase inhibitors

Li¹,

Wang²,

Tanizaki

et al. 2014

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

163

132

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The human FGF receptors (FGFRs) play critical roles in various human cancers, and several FGFR inhibitors are currently under clinical investigation. Resistance usually results from selection for mutant kinases that are impervious to the action of the drug or from up-regulation of compensatory signaling pathways. Preclinical studies have demonstrated that resistance to FGFR inhibitors can be acquired through mutations in the FGFR gatekeeper residue, as clinically observed for FGFR4 in embryonal rhabdomyosarcoma and neuroendocrine breast carcinomas. Here we report on the use of a structure-based drug design to develop two selective, next-generation covalent FGFR inhibitors, the FGFR irreversible inhibitors 2 (FIIN-2) and 3 (FIIN-3). To our knowledge, FIIN-2 and FIIN-3 are the first inhibitors that can potently inhibit the proliferation of cells dependent upon the gatekeeper mutants of FGFR1 or FGFR2, which confer resistance to first-generation clinical FGFR inhibitors such as NVP-BGJ398 and AZD4547. Because of the conformational flexibility of the reactive acrylamide substituent, FIIN-3 has the unprecedented ability to inhibit both the EGF receptor (EGFR) and FGFR covalently by targeting two distinct cysteine residues. We report the cocrystal structure of FGFR4 with FIIN-2, which unexpectedly exhibits a "DFG-out" covalent binding mode. The structural basis for dual FGFR and EGFR targeting by FIIN3 also is illustrated by crystal structures of FIIN-3 bound with FGFR4 V550L and EGFR L858R. These results have important implications for the design of covalent FGFR inhibitors that can overcome clinical resistance and provide the first example, to our knowledge, of a kinase inhibitor that covalently targets cysteines located in different positions within the ATP-binding pocket.drug discovery | cancer drug resistance | kinase inhibitor | structure-based drug design R eceptor tyrosine kinases (RTKs) serve as critical sensors of extracellular cues that activate a myriad of intracellular signaling pathways to regulate cell state. There are 58 receptor tyrosine kinases in the human genome, and many have been demonstrated to be constitutively activated through amplification or mutation in particular cancers. The signals emanating from these RTKs, such as epidermal growth factor receptor (EGFR), FGF receptor (FGFR), platelet-derived growth factor receptor (PDGFR), protein kinase Kit (KIT), and protein kinase c-Met (MET), have been pharmacologically proven to be essential to the survival of cancers expressing mutant forms of these proteins. However, rapid resistance to monotherapy with first-generation RTK inhibitors has been universally observed. Resistance typically arises from the emergence of cancer cells expressing mutant forms of RTKs that are impervious to the action of first-generation drugs or from the activation of by-pass signaling mechanisms. Resistance can be overcome by developing new inhibitors that target the mutant RTK directly or target bypass signaling mechanisms. Indeed this approach has been deployed succes...

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High-Throughput Kinase Profiling: A More Efficient Approach toward the Discovery of New Kinase Inhibitors

Cited by 108 publications

References 55 publications

A genome-wide RNA interference screen identifies new regulators of androgen receptor function in prostate cancer cells

A genome-wide RNA interference screen identifies new regulators of androgen receptor function in prostate cancer cells

PIM Kinases Are Essential for Chronic Lymphocytic Leukemia Cell Survival (PIM2/3) and CXCR4-Mediated Microenvironmental Interactions (PIM1)

Development of covalent inhibitors that can overcome resistance to first-generation FGFR kinase inhibitors

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