Identification and Characterization of JAK2 Pseudokinase Domain Small Molecule Binders

Puleo, David E.; Kucera, Kaury; Hammarén, Henrik M.; Ungureanu, Daniela; Newton, Ana S.; Silvennoinen, Olli; Jorgensen, William L.; Schlessinger, Joseph

doi:10.1021/acsmedchemlett.7b00153

Title: Identification and Characterization of JAK2 Pseudokinase Domain Small Molecule Binders

Journal Article · Mon May 22 00:00:00 EDT 2017 · ACS Medicinal Chemistry Letters

DOI:https://doi.org/10.1021/acsmedchemlett.7b00153· OSTI ID:1390880

Puleo, David E. ^[1]; Kucera, Kaury ^[1]; Hammarén, Henrik M. ^[2]; Ungureanu, Daniela ^[2]; Newton, Ana S. ^[1]; Silvennoinen, Olli ^[2];

^[1];

^[1]

Yale Univ., New Haven, CT (United States)
Tampere Univ. of Technology (Finland)

Janus kinases (JAKs) regulate hematopoiesis via the cytokine-mediated JAK-STAT signaling pathway. JAKs contain tandem C-terminal pseudokinase (JH2) and tyrosine kinase (JH1) domains. The JAK2 pseudokinase domain adopts a protein kinase fold and, despite its pseudokinase designation, binds ATP with micromolar affinity. Recent evidence shows that displacing ATP from the JAK2 JH2 domain alters the hyperactivation state of the oncogenic JAK2 V617F protein while sparing the wild type JAK2 protein. In this study, small molecule binders of JAK2 JH2 were identified via an in vitro screen. Top hits were characterized using biophysical and structural approaches. Development of pseudokinase-selective compounds may offer novel pharmacological opportunities for treating cancers driven by JAK2 V617F and other oncogenic JAK mutants.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: 1S10OD018007

OSTI ID:: 1390880

Journal Information:: ACS Medicinal Chemistry Letters, Vol. 8, Issue 6; ISSN 1948-5875

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 28 works

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References (19)

The JAK-STAT Pathway: Impact on Human Disease and Therapeutic Intervention O'Shea, John J.; Schwartz, Daniella M.; Villarino, Alejandro V. Annual Review of Medicine, Vol. 66, Issue 1 https://doi.org/10.1146/annurev-med-051113-024537	journal	January 2015
Regulation of the Jak2 Tyrosine Kinase by Its Pseudokinase Domain Saharinen, Pipsa; Takaluoma, Kati; Silvennoinen, Olli Molecular and Cellular Biology, Vol. 20, Issue 10 https://doi.org/10.1128/MCB.20.10.3387-3395.2000	journal	May 2000
Structure of the pseudokinase-kinase domains from protein kinase TYK2 reveals a mechanism for Janus kinase (JAK) autoinhibition Lupardus, P. J.; Ultsch, M.; Wallweber, H. Proceedings of the National Academy of Sciences, Vol. 111, Issue 22 https://doi.org/10.1073/pnas.1401180111	journal	May 2014
Molecular basis for pseudokinase-dependent autoinhibition of JAK2 tyrosine kinase Shan, Yibing; Gnanasambandan, Kavitha; Ungureanu, Daniela Nature Structural & Molecular Biology, Vol. 21, Issue 7 https://doi.org/10.1038/nsmb.2849	journal	June 2014
Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders Baxter, E.; Scott, L.; Campbell, P. The Lancet, Vol. 365, Issue 9464 https://doi.org/10.1016/S0140-6736(05)74230-6	journal	March 2005
A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera James, Chloé; Ugo, Valérie; Le Couédic, Jean-Pierre Nature, Vol. 434, Issue 7037 https://doi.org/10.1038/nature03546	journal	March 2005
A Gain-of-Function Mutation of JAK2 in Myeloproliferative Disorders Kralovics, Robert; Passamonti, Francesco; Buser, Andreas S. New England Journal of Medicine, Vol. 352, Issue 17 https://doi.org/10.1056/NEJMoa051113	journal	April 2005
Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis Levine, Ross L.; Wadleigh, Martha; Cools, Jan Cancer Cell, Vol. 7, Issue 4 https://doi.org/10.1016/j.ccr.2005.03.023	journal	April 2005
Structure of a pseudokinase-domain switch that controls oncogenic activation of Jak kinases Toms, Angela V.; Deshpande, Anagha; McNally, Randall Nature Structural & Molecular Biology, Vol. 20, Issue 10 https://doi.org/10.1038/nsmb.2673	journal	September 2013
The pseudokinase domain of JAK2 is a dual-specificity protein kinase that negatively regulates cytokine signaling Ungureanu, Daniela; Wu, Jinhua; Pekkala, Tuija Nature Structural & Molecular Biology, Vol. 18, Issue 9 https://doi.org/10.1038/nsmb.2099	journal	August 2011
Crystal structures of the JAK2 pseudokinase domain and the pathogenic mutant V617F Bandaranayake, Rajintha M.; Ungureanu, Daniela; Shan, Yibing Nature Structural & Molecular Biology, Vol. 19, Issue 8 https://doi.org/10.1038/nsmb.2348	journal	July 2012
Structural and Functional Characterization of the JH2 Pseudokinase Domain of JAK Family Tyrosine Kinase 2 (TYK2) Min, Xiaoshan; Ungureanu, Daniela; Maxwell, Sarah Journal of Biological Chemistry, Vol. 290, Issue 45 https://doi.org/10.1074/jbc.M115.672048	journal	November 2015
ATP binding to the pseudokinase domain of JAK2 is critical for pathogenic activation Hammarén, Henrik M.; Ungureanu, Daniela; Grisouard, Jean Proceedings of the National Academy of Sciences, Vol. 112, Issue 15 https://doi.org/10.1073/pnas.1423201112	journal	March 2015
1-Acyl-1 H -[1,2,4]triazole-3,5-diamine Analogues as Novel and Potent Anticancer Cyclin-Dependent Kinase Inhibitors: Synthesis and Evaluation of Biological Activities Lin, Ronghui; Connolly, Peter J.; Huang, Shenlin Journal of Medicinal Chemistry, Vol. 48, Issue 13 https://doi.org/10.1021/jm050267e	journal	June 2005
A quantitative analysis of kinase inhibitor selectivity Karaman, Mazen W.; Herrgard, Sanna; Treiber, Daniel K. Nature Biotechnology, Vol. 26, Issue 1 https://doi.org/10.1038/nbt1358	journal	January 2008
Fragment-Based Discovery of the Pyrazol-4-yl Urea (AT9283), a Multitargeted Kinase Inhibitor with Potent Aurora Kinase Activity ^† Howard, Steven; Berdini, Valerio; Boulstridge, John A. Journal of Medicinal Chemistry, Vol. 52, Issue 2 https://doi.org/10.1021/jm800984v	journal	January 2009
JAK2 V617F Constitutive Activation Requires JH2 Residue F595: A Pseudokinase Domain Target for Specific Inhibitors Dusa, Alexandra; Mouton, Céline; Pecquet, Christian PLoS ONE, Vol. 5, Issue 6 https://doi.org/10.1371/journal.pone.0011157	journal	June 2010
Autoinhibition of Jak2 Tyrosine Kinase Is Dependent on Specific Regions in Its Pseudokinase Domain Saharinen, Pipsa; Vihinen, Mauno; Silvennoinen, Olli Molecular Biology of the Cell, Vol. 14, Issue 4 https://doi.org/10.1091/mbc.e02-06-0342	journal	April 2003
Regulation of the Jak2 Tyrosine Kinase by Its Pseudokinase Domain Saharinen, Pipsa; Takaluoma, Kati; Silvennoinen, Olli Molecular and Cellular Biology, Vol. 20, Issue 10 https://doi.org/10.1128/.20.10.3387-3395.2000	journal	January 2000

Cited By (6)

Covalent inhibitors of EGFR family protein kinases induce degradation of human Tribbles 2 (TRIB2) pseudokinase in cancer cells Foulkes, Daniel M.; Byrne, Dominic P.; Yeung, Wayland Science Signaling, Vol. 11, Issue 549 https://doi.org/10.1126/scisignal.aat7951	journal	September 2018
Prospects for pharmacological targeting of pseudokinases Kung, Jennifer E.; Jura, Natalia Nature Reviews Drug Discovery https://doi.org/10.1038/s41573-019-0018-3	journal	March 2019
Janus kinases to jakinibs: from basic insights to clinical practice Gadina, Massimo; Le, Mimi T.; Schwartz, Daniella M. Rheumatology, Vol. 58, Issue Supplement_1 https://doi.org/10.1093/rheumatology/key432	journal	February 2019
The PEAK family of pseudokinases, their role in cell signalling and cancer Patel, Onisha; Roy, Michael J.; Murphy, James M. The FEBS Journal, Vol. 287, Issue 19 https://doi.org/10.1111/febs.15087	journal	November 2019
JAK inhibitors for the treatment of myeloproliferative neoplasms and other disorders Vainchenker, William; Leroy, Emilie; Gilles, Laure F1000Research, Vol. 7 https://doi.org/10.12688/f1000research.13167.1	journal	January 2018
Repurposing covalent EGFR/HER2 inhibitors for on-target degradation of human Tribbles 2 (TRIB2) pseudokinase Foulkes, Daniel M.; Byrne, Dominic P.; Bailey, Fiona P. https://doi.org/10.1101/305243	posted_content	April 2018

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59 BASIC BIOLOGICAL SCIENCES
JAK2
Pseudokinase
JH2
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