New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation

Matthew L. Arwood; Yao Liu; Shannon K. Harkins; David M. Weinstock; Lei Yang; Kristen E. Stevenson; Olivia D. Plana; Jingyun Dong; Haley Cirka; Kristen L. Jones; Anniina T. Virtanen; Dikshat G. Gupta; Amanda Ceas; Brian Lawney; Akinori Yoda; Catharine Leahy; Mingfeng Hao; Zhixiang He; Hwan Geun Choi; Yaning Wang; Olli Silvennoinen; Stevan R. Hubbard; Tinghu Zhang; Nathanael S. Gray; Loretta S. Li

doi:10.1016/j.chembiol.2023.05.007

New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation

Matthew L. Arwood, Yao Liu, Shannon K. Harkins, David M. Weinstock, Lei Yang, Kristen E. Stevenson, Olivia D. Plana, Jingyun Dong, Haley Cirka, Kristen L. Jones, Anniina T. Virtanen, Dikshat G. Gupta, Amanda Ceas, Brian Lawney, Akinori Yoda, Catharine Leahy, Mingfeng Hao, Zhixiang He, Hwan Geun Choi, Yaning WangOlli Silvennoinen, Stevan R. Hubbard, Tinghu Zhang, Nathanael S. Gray^*, Loretta S. Li^*

^*Corresponding author for this work

Pediatrics

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

Recurrent JAK2 alterations are observed in myeloproliferative neoplasms, B-cell acute lymphoblastic leukemia, and other hematologic malignancies. Currently available type I JAK2 inhibitors have limited activity in these diseases. Preclinical data support the improved efficacy of type II JAK2 inhibitors, which lock the kinase in the inactive conformation. By screening small molecule libraries, we identified a lead compound with JAK2 selectivity. We highlight analogs with on-target biochemical and cellular activity and demonstrate in vivo activity using a mouse model of polycythemia vera. We present a co-crystal structure that confirms the type II binding mode of our compounds with the “DFG-out” conformation of the JAK2 activation loop. Finally, we identify a JAK2 G993A mutation that confers resistance to the type II JAK2 inhibitor CHZ868 but not to our analogs. These data provide a template for identifying novel type II kinase inhibitors and inform further development of agents targeting JAK2 that overcome resistance.

Original language	English (US)
Pages (from-to)	618-631.e12
Journal	Cell Chemical Biology
Volume	30
Issue number	6
DOIs	https://doi.org/10.1016/j.chembiol.2023.05.007
State	Published - Jun 15 2023

Funding

This work was supported by: the Prostate Cancer Foundation (N.S.G., S.R.H., O.S., and D.M.W.), the Damon Runyon Physician-Scientist Training Award (L.S.L.), CureSearch for Children’s Cancer (L.S.L.), the Rally! Foundation for Childhood Cancer Research (L.S.L.), a Conquer Cancer Young Investigator Award funded by the Strike 3 Foundation (L.S.L.), an Institutional Research Grant , IRG-21-144-27 , from the American Cancer Society (L.S.L.), the Stanley Manne Children’s Research Institute and the Ann & Robert H. Lurie Children’s Hospital of Chicago (L.S.L.), Academy of Finland (A.T.V. and O.S.), Sigrid Jusélius Foundation (A.T.V. and O.S.), Cancer Foundation Finland (A.T.V. and O.S.), Jane and Aatos Erkko Foundation (A.T.V. and O.S.), Tampere Tuberculosis Foundation (A.T.V. and O.S.), and Pirkanmaa hospital district competitive research funding (A.T.V. and O.S.). We thank the Robert H. Lurie Comprehensive Cancer Center of Northwestern University in Chicago, IL for the use of the Flow Cytometry Core Facility. The Lurie Cancer Center is supported in part by an NCI Cancer Center Support Grant #P30 CA060553. The authors thank Renee Rubio from the Dana-Farber Cancer Institute Center for Cancer Computational Biology for technical assistance with the RNA sequencing. Portions of the graphical abstract (QQ23JH07KJ) and Figure 4 A (KO23JGZX77) were created with BioRender.com .

Keywords

B-ALL
CHZ868
CRLF2
JAK2
kinase
myeloproliferative neoplasm
resistance
ruxolitinib
type II inhibitor

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.chembiol.2023.05.007

Cite this

Arwood, M. L., Liu, Y., Harkins, S. K., Weinstock, D. M., Yang, L., Stevenson, K. E., Plana, O. D., Dong, J., Cirka, H., Jones, K. L., Virtanen, A. T., Gupta, D. G., Ceas, A., Lawney, B., Yoda, A., Leahy, C., Hao, M., He, Z., Choi, H. G., ... Li, L. S. (2023). New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation. Cell Chemical Biology, 30(6), 618-631.e12. https://doi.org/10.1016/j.chembiol.2023.05.007

@article{31c872ad758e443894d6d4ccbf6c8864,

title = "New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation",

abstract = "Recurrent JAK2 alterations are observed in myeloproliferative neoplasms, B-cell acute lymphoblastic leukemia, and other hematologic malignancies. Currently available type I JAK2 inhibitors have limited activity in these diseases. Preclinical data support the improved efficacy of type II JAK2 inhibitors, which lock the kinase in the inactive conformation. By screening small molecule libraries, we identified a lead compound with JAK2 selectivity. We highlight analogs with on-target biochemical and cellular activity and demonstrate in vivo activity using a mouse model of polycythemia vera. We present a co-crystal structure that confirms the type II binding mode of our compounds with the “DFG-out” conformation of the JAK2 activation loop. Finally, we identify a JAK2 G993A mutation that confers resistance to the type II JAK2 inhibitor CHZ868 but not to our analogs. These data provide a template for identifying novel type II kinase inhibitors and inform further development of agents targeting JAK2 that overcome resistance.",

keywords = "B-ALL, CHZ868, CRLF2, JAK2, kinase, myeloproliferative neoplasm, resistance, ruxolitinib, type II inhibitor",

author = "Arwood, {Matthew L.} and Yao Liu and Harkins, {Shannon K.} and Weinstock, {David M.} and Lei Yang and Stevenson, {Kristen E.} and Plana, {Olivia D.} and Jingyun Dong and Haley Cirka and Jones, {Kristen L.} and Virtanen, {Anniina T.} and Gupta, {Dikshat G.} and Amanda Ceas and Brian Lawney and Akinori Yoda and Catharine Leahy and Mingfeng Hao and Zhixiang He and Choi, {Hwan Geun} and Yaning Wang and Olli Silvennoinen and Hubbard, {Stevan R.} and Tinghu Zhang and Gray, {Nathanael S.} and Li, {Loretta S.}",

note = "Publisher Copyright: {\textcopyright} 2023 Elsevier Ltd",

year = "2023",

month = jun,

day = "15",

doi = "10.1016/j.chembiol.2023.05.007",

language = "English (US)",

volume = "30",

pages = "618--631.e12",

journal = "Cell Chemical Biology",

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Arwood, ML, Liu, Y, Harkins, SK, Weinstock, DM, Yang, L, Stevenson, KE, Plana, OD, Dong, J, Cirka, H, Jones, KL, Virtanen, AT, Gupta, DG, Ceas, A, Lawney, B, Yoda, A, Leahy, C, Hao, M, He, Z, Choi, HG, Wang, Y, Silvennoinen, O, Hubbard, SR, Zhang, T, Gray, NS & Li, LS 2023, 'New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation', Cell Chemical Biology, vol. 30, no. 6, pp. 618-631.e12. https://doi.org/10.1016/j.chembiol.2023.05.007

TY - JOUR

T1 - New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation

AU - Arwood, Matthew L.

AU - Liu, Yao

AU - Harkins, Shannon K.

AU - Weinstock, David M.

AU - Yang, Lei

AU - Stevenson, Kristen E.

AU - Plana, Olivia D.

AU - Dong, Jingyun

AU - Cirka, Haley

AU - Jones, Kristen L.

AU - Virtanen, Anniina T.

AU - Gupta, Dikshat G.

AU - Ceas, Amanda

AU - Lawney, Brian

AU - Yoda, Akinori

AU - Leahy, Catharine

AU - Hao, Mingfeng

AU - He, Zhixiang

AU - Choi, Hwan Geun

AU - Wang, Yaning

AU - Silvennoinen, Olli

AU - Hubbard, Stevan R.

AU - Zhang, Tinghu

AU - Gray, Nathanael S.

AU - Li, Loretta S.

PY - 2023/6/15

Y1 - 2023/6/15

N2 - Recurrent JAK2 alterations are observed in myeloproliferative neoplasms, B-cell acute lymphoblastic leukemia, and other hematologic malignancies. Currently available type I JAK2 inhibitors have limited activity in these diseases. Preclinical data support the improved efficacy of type II JAK2 inhibitors, which lock the kinase in the inactive conformation. By screening small molecule libraries, we identified a lead compound with JAK2 selectivity. We highlight analogs with on-target biochemical and cellular activity and demonstrate in vivo activity using a mouse model of polycythemia vera. We present a co-crystal structure that confirms the type II binding mode of our compounds with the “DFG-out” conformation of the JAK2 activation loop. Finally, we identify a JAK2 G993A mutation that confers resistance to the type II JAK2 inhibitor CHZ868 but not to our analogs. These data provide a template for identifying novel type II kinase inhibitors and inform further development of agents targeting JAK2 that overcome resistance.

AB - Recurrent JAK2 alterations are observed in myeloproliferative neoplasms, B-cell acute lymphoblastic leukemia, and other hematologic malignancies. Currently available type I JAK2 inhibitors have limited activity in these diseases. Preclinical data support the improved efficacy of type II JAK2 inhibitors, which lock the kinase in the inactive conformation. By screening small molecule libraries, we identified a lead compound with JAK2 selectivity. We highlight analogs with on-target biochemical and cellular activity and demonstrate in vivo activity using a mouse model of polycythemia vera. We present a co-crystal structure that confirms the type II binding mode of our compounds with the “DFG-out” conformation of the JAK2 activation loop. Finally, we identify a JAK2 G993A mutation that confers resistance to the type II JAK2 inhibitor CHZ868 but not to our analogs. These data provide a template for identifying novel type II kinase inhibitors and inform further development of agents targeting JAK2 that overcome resistance.

KW - B-ALL

KW - CHZ868

KW - CRLF2

KW - JAK2

KW - kinase

KW - myeloproliferative neoplasm

KW - resistance

KW - ruxolitinib

KW - type II inhibitor

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UR - http://www.scopus.com/inward/citedby.url?scp=85163098688&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2023.05.007

DO - 10.1016/j.chembiol.2023.05.007

M3 - Article

C2 - 37290440

AN - SCOPUS:85163098688

SN - 2451-9456

VL - 30

SP - 618-631.e12

JO - Cell Chemical Biology

JF - Cell Chemical Biology

IS - 6

ER -

New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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Crystal structure of JAK2 JH1 with type II inhibitor YLIU-04-105-1

Cite this

New scaffolds for type II JAK2 inhibitors overcome the acquired G993A resistance mutation

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Datasets

Crystal structure of JAK2 JH1 with type II inhibitor YLIU-04-105-1

Cite this