Selective Covalent Targeting of Pyruvate Kinase M2 Using Arsenous Warheads
Abstract
There is growing interest in covalent targeted inhibitors in drug discovery against previously “undruggable” sites and targets. These molecules typically feature an electrophilic warhead that reacts with nucleophilic groups of protein residues, most notably the thiol group of cysteines. One main challenge in the field is to develop versatile utilizable warheads. Here, we characterize the unique features of novel arsenous warheads for reaction with thiol species in a reversible manner and further demonstrate that organoarsenic probes can be chemically tuned toward specific molecular targets by developing selective and potent inhibitors of pyruvate kinase M2 (PKM2). We show that compound 24 is a covalent and allosteric inhibitor of PKM2 and its orally bioavailable prodrug 25 exerts efficacious inhibition of PKM2-dependent tumor growth in vitro and in vivo . Our results introduce 25 and its derivatives as useful pharmacological tools and provide a general road map for targeting the protein cysteinome using arsenous warheads.
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A novel PKM2 inhibitor (compound 78) based on trivalent arsine covalent warheads was reported.
“In a recent study on targeted covalent inhibitors, the Cross Center of Shanghai Institute of Organic Sciences and collaborators reported a novel PKM2 inhibitor, compound (78) (Table 1), based on trivalent arsine covalent warheads.”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [229]
Cites
Canonical dynamics: Equilibrium phase-space distributions
Hoover, W. G. (1985) · Physical Review A · doi:10.1103/physreva.31.1695 · Cited by 23,453 · see citing works
Constant pressure molecular dynamics algorithms
Martyna, G. J., Tobias, D. J., & Klein, M. L. (1994) · The Journal of Chemical Physics · doi:10.1063/1.467468 · Cited by 5,845 · see citing works
OPLS3: A Force Field Providing Broad Coverage of Drug-like Small Molecules and Proteins.
Harder, E. D., Damm, W., Maple, J., Wu, C., Reboul, M., Xiang, J.-Y., Wang, L.-L., Lupyan, D., Dahlgren, M. K., Knight, J. L., et al. (2016) · Journal of Chemical Theory and Computation · doi:10.1021/acs.jctc.5b00864 · Cited by 2,826 · see citing works
Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay
Molina, D. M., Jafari, R., Ignatushchenko, M., Seki, T., Larsson, E. A., Dan, C., Sreekumar, L., Cao, Y., & Nordlund, P. (2013) · Science · doi:10.1126/science.1233606 · Cited by 2,261 · see citing works
The resurgence of covalent drugs
Singh, J., Petter, R. C., Baillie, T. A., & Whitty, A. (2011) · Nature Reviews Drug Discovery · doi:10.1038/nrd3410 · Cited by 1,873 · see citing works
Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells
Griffin, B. A., Adams, S., & Tsien, R. Y. (1998) · Science · doi:10.1126/science.281.5374.269 · Cited by 1,553 · see citing works
Rethinking ovarian cancer: recommendations for improving outcomes
Vaughan, S., Coward, J. I. G., Bast, R. C., Berchuck, A., Berek, J. S., Brenton, J. D., Coukos, G., Crum, C. P., Drapkin, R., Etemadmoghadam, D., et al. (2011) · Nature reviews. Cancer · doi:10.1038/nrc3144 · Cited by 1,470 · see citing works
Proteome-wide covalent ligand discovery in native biological systems
Backus, K. M., Correia, B. E., Lum, K. M., Forli, S., Horning, B. D., González-Páez, G. E., Chatterjee, S., Lanning, B. R., Teijaro, J. R., Olson, A. J., et al. (2016) · Nature · doi:10.1038/nature18002 · Cited by 940 · see citing works
Shen, S., Li, X., Cullen, W., Weinfeld, M., & Le, X. C. (2013) · Chemical Reviews · doi:10.1021/cr300015c · Cited by 822 · see citing works
Pyruvate Kinase M2 Regulates Gene Transcription by Acting as a Protein Kinase
Gao, X., Wang, H., Yang, J. J., Liu, X., & Liu, Z.-R. (2012) · Molecular Cell · doi:10.1016/j.molcel.2012.01.001 · Cited by 772 · see citing works
Fell, J. B., Fischer, J. P., Baer, B. R., Blake, J. F., Bouhana, K., Briere, D. M., Brown, K. D., Burgess, L. E., Burns, A. C., Burkard, M. R., et al. (2020) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.9b02052 · Cited by 653 · see citing works
PKM2-dependent glycolysis promotes NLRP3 and AIM2 inflammasome activation
Xie, M., Yu, Y., Kang, R., Zhu, S., Yang, L., Zeng, L., Sun, X., Yang, M., Billiar, T. R., Wang, H., et al. (2016) · Nature Communications · doi:10.1038/ncomms13280 · Cited by 567 · see citing works
Shikonin and its analogs inhibit cancer cell glycolysis by targeting tumor pyruvate kinase-M2
Chen, J., Xie, J., Jiang, Z., Wang, B., Wang, Y., & Hu, X. (2011) · Oncogene · doi:10.1038/onc.2011.137 · Cited by 515 · see citing works
PKM2 regulates the Warburg effect and promotes HMGB1 release in sepsis
Yang, L., Xie, M., Yang, M., Yu, Y., Zhu, S., Hou, W., Kang, R., Lotze, M. T., Billiar, T. R., Wang, H., et al. (2014) · Nature Communications · doi:10.1038/ncomms5436 · Cited by 503 · see citing works
Structure-based design of targeted covalent inhibitors
Lonsdale, R., & Ward, R. A. (2018) · Chemical Society Reviews · doi:10.1039/c7cs00220c · Cited by 344 · see citing works
Flanagan, M. E., Abramite, J. A., Anderson, D. P., AULABAUGH, A. E., Dahal, U. P., Gilbert, A. M., Li, C., Montgomery, J. I., Oppenheimer, S. R., Ryder, T. F., et al. (2014) · Journal of Medicinal Chemistry · doi:10.1021/jm501412a · Cited by 337 · see citing works
Rapid Covalent-Probe Discovery by Electrophile-Fragment Screening
Resnick, E., Bradley, A., Gan, J., Douangamath, A., Krojer, T., Sethi, R., Geurink, P. P., Aimon, A., Amitai, G., Bellini, D., et al. (2019) · Journal of the American Chemical Society · doi:10.1021/jacs.9b02822 · Cited by 335 · see citing works
How acute promyelocytic leukaemia revived arsenic
Zhu, J., Chen, Z., Lallemand-Breitenbach, V., & de Thé, H. (2002) · Nature reviews. Cancer · doi:10.1038/nrc887 · Cited by 324 · see citing works
Covalent docking of large libraries for the discovery of chemical probes
London, N., Miller, R. M., Krishnan, S., Uchida, K., Irwin, J. J., Eidam, O., Gibold, L., Cimermančič, P., Bonnet, R., Shoichet, B. K., et al. (2014) · Nature Chemical Biology · doi:10.1038/nchembio.1666 · Cited by 274 · see citing works
Identification of small molecule inhibitors of pyruvate kinase M2
Heiden, M. G. V., Christofk, H. R., Schuman, E., Subtelny, A. O., Sharfi, H., Harlow, E. E., Xian, J., & Cantley, L. C. (2009) · Biochemical Pharmacology · doi:10.1016/j.bcp.2009.12.003 · Cited by 239 · see citing works
Pyruvate Kinase M2: Multiple Faces for Conferring Benefits on Cancer Cells
Tamada, M., Suematsu, M., & Saya, H. (2012) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-12-0859 · Cited by 239 · see citing works
The Cysteinome of Protein Kinases as a Target in Drug Development
Chaikuad, A., Koch, P., Laufer, S., & Knapp, S. (2017) · Angewandte Chemie International Edition · doi:10.1002/anie.201707875 · Cited by 237 · see citing works
Pyruvate kinase M2-specific siRNA induces apoptosis and tumor regression
Goldberg, M. S., & Sharp, P. A. (2012) · The Journal of Experimental Medicine · doi:10.1084/jem.20111487 · Cited by 232 · see citing works
Chen, X., Yang, L., Zhang, N., Turpin, J. A., Buckheit, R. W., Osterling, C., Oppenheim, J. J., & Howard, O. M. Z. (2003) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.47.9.2810-2816.2003 · Cited by 226 · see citing works
Chen, S., Zhou, G., Zhang, X., Mao, J.-H., de Thé, H., & Chen, Z. (2011) · Blood · doi:10.1182/blood-2010-11-283598 · Cited by 184 · see citing works
M2 pyruvate kinase provides a mechanism for nutrient sensing and regulation of cell proliferation
Morgan, H. P., O’Reilly, F. J., Wear, M. A., O’Neill, J. R., Fothergill‐Gilmore, L. A., Hupp, T. R., & Walkinshaw, M. D. (2013) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1217157110 · Cited by 180 · see citing works
Duan, D., Zhang, B., Yao, J., Liu, Y., & Fang, J. (2014) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2014.02.016 · Cited by 175 · see citing works
Natural Product Micheliolide (MCL) Irreversibly Activates Pyruvate Kinase M2 and Suppresses Leukemia
Li, J., Li, S., Guo, J., Li, Q., Long, J., Ma, C., Ding, Y., Yan, C., Li, L., Wu, Z., et al. (2018) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.8b00241 · Cited by 154 · see citing works
Activatable Mitochondria‐Targeting Organoarsenic Prodrugs for Bioenergetic Cancer Therapy
Luo, X., Gong, X., Su, L., Lin, H., Yang, Z., Yan, X., & Gao, J. (2020) · Angewandte Chemie International Edition · doi:10.1002/anie.202012237 · Cited by 128 · see citing works
Melarsoprol Resistance in African Trypanosomiasis
Fairlamb, A. H., & Horn, D. (2018) · Trends in Parasitology · doi:10.1016/j.pt.2018.04.002 · Cited by 125 · see citing works
2-Sulfonylpyridines as Tunable, Cysteine-Reactive Electrophiles
Zambaldo, C., Vinogradova, E. V., Qi, X., Iaconelli, J., Suciu, R. M., Koh, M., Senkane, K., Chadwick, S. R., Sanchez, B. B., Chen, J. S., et al. (2020) · Journal of the American Chemical Society · doi:10.1021/jacs.0c02721 · Cited by 117 · see citing works
Ning, X., Qi, H., Li, R., Li, Y., Jin, Y., McNutt, M. A., Liu, J., & Yin, Y. (2017) · European Journal of Medicinal Chemistry · doi:10.1016/j.ejmech.2017.06.064 · Cited by 115 · see citing works
Self-citationDual Inhibition of CDK12/CDK13 Targets Both Tumor and Immune Cells in Ovarian Cancer
Cheng, L., Zhou, S., Zhou, S., Shi, K., Cheng, Y., Cai, M., Ye, K., Lin, L., Zhang, Z., Jia, C., et al. (2022) · Cancer Research · doi:10.1158/0008-5472.can-22-0222 · Cited by 54 · see citing works
Self-citationShikonin Inhibits Intestinal Calcium-Activated Chloride Channels and Prevents Rotaviral Diarrhea
Jiang, Y., Yu, B., Yang, H. Y., & Ma, T. (2016) · Frontiers in Pharmacology · doi:10.3389/fphar.2016.00270 · Cited by 45 · see citing works
Covalent binders in drug discovery
Vasudevan, A., Argiriadi, M. A., Baranczak, A., Friedman, M., Gavrilyuk, J. I., Hobson, A. D., Hulce, J. J., Osman, S., & Wilson, N. S. (2019) · Progress in medicinal chemistry · doi:10.1016/bs.pmch.2018.12.002 · Cited by 42 · see citing works
SOX17 and PAX8 constitute an actionable lineage-survival transcriptional complex in ovarian cancer
Lin, L., Shi, K., Zhou, S., Cai, M., Zhang, C., Sun, Y., Zang, J., Cheng, L., Ye, K., Ma, P., et al. (2022) · Oncogene · doi:10.1038/s41388-022-02210-3 · Cited by 31 · see citing works
Self-citationDevelopment of Novel Irreversible Pyruvate Kinase M2 Inhibitors
Hsieh, I., Gopula, B., Chou, C., Wu, H.-Y., Chang, G., Wu, W., Chang, C., Chu, P., & Chen, C. (2019) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.9b00763 · Cited by 28 · see citing works
Frampton, J. E. (2022) · Drugs · doi:10.1007/s40265-022-01795-z · Cited by 19 · see citing works
Golemović, M., Quintás‐Cardama, A., Manshouri, T., Oršolić, N., Duzkale, H., Johansen, M. J., Freireich, E. J., Kantarjian, H. M., Zingaro, R. A., & Verstovšek, S. (2009) · Investigational New Drugs · doi:10.1007/s10637-009-9267-z · Cited by 17 · see citing works
Flagged first, then by citations
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Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials
Xie, X., Yu, T., Li, X., Zhang, N., Foster, L. J., Peng, C., Huang, W., & He, G. (2023) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-023-01589-z · Cited by 396 · see citing works
Self-citationThe expanding repertoire of covalent warheads for drug discovery
Mehta, N. V., & Degani, M. S. (2023) · Drug Discovery Today · doi:10.1016/j.drudis.2023.103799 · Cited by 72 · see citing works
Glutathione‐triggered prodrugs: Design strategies, potential applications, and perspectives
Zhao, J., Li, X., Ma, T., Chang, B., Zhang, B., & Fang, J. (2023) · Medicinal Research Reviews · doi:10.1002/med.22007 · Cited by 53 · see citing works
Das, R., Pulugu, P., Singh, A. A., Chatterjee, D. R., Baviskar, S., Vyas, H., Behera, S. K., Srivastava, A., Kumar, H., & Shard, A. (2024) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.3c01512 · Cited by 27 · see citing works
Pretargeted Mitochondrial Delivery of Organoarsenicals for Cancer Immunotherapy
Wang, R., Tian, Y., Lü, X., Fang, L., Miao, Y., Fang, D., Li, Y., Liu, H. S., & Ye, D. (2025) · Journal of the American Chemical Society · doi:10.1021/jacs.5c12201 · Cited by 19 · see citing works
Self-citationA Pyroptosis‐Inducing Arsenic(III) Nanomicelle Platform for Synergistic Cancer Immunotherapy
Wang, X., Tang, Y., Li, Y., & Qi, Z. (2024) · Advanced Healthcare Materials · doi:10.1002/adhm.202401904 · Cited by 14 · see citing works
Self-citationZhao, Y., Zhao, X., Duan, L., Hou, R., Gu, Y., Liu, Z., Chen, J., Wu, F., Yang, L., Le, X. C., et al. (2024) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.3c02076 · Cited by 11 · see citing works
Chowdhury, M. G., Singh, A. A., Bhattacharyya, M., Muthukumar, V., Kapoor, S., Srivastava, A., Kumar, H., & Shard, A. (2025) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.5c02169 · Cited by 6 · see citing works
Recent progress in the development of small molecule pyruvate kinase M2 inhibitors: 2020–2025
Das, R., Sharma, S. K., Ambast, P. K., & Shard, A. (2025) · Future Medicinal Chemistry · doi:10.1080/17568919.2025.2571029 · Cited by 5 · see citing works
Wang, X., Qiu, J., Xie, J., Huang, R., Chen, Y., Fang, Y., Tang, Y., Wu, S., Zhou, X., Chang, Z., et al. (2025) · Bioorganic Chemistry · doi:10.1016/j.bioorg.2025.109212 · Cited by 2 · see citing works
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Castelôa, M., Borges, F., Benfeito, S., & Chavarria, D. (2026) · European Journal of Medicinal Chemistry · doi:10.1016/j.ejmech.2026.119264 · Cited by 2 · see citing works
Bai, S., Gao, M., You, Q., & Wang, L. (2026) · European Journal of Medicinal Chemistry · doi:10.1016/j.ejmech.2026.119314 · Cited by 1 · see citing works
Li, W., Shi, Y., Chen, X., Wang, H., Wei, D., Yao, J., Li, X., Lü, J., Li, X., Chang, J., et al. (2025) · Biochemical Pharmacology · doi:10.1016/j.bcp.2025.116997 · Cited by 1 · see citing works
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- Authors
- Wang, J., Zhou, S., Cheng, Y., Cheng, L., Qin, Y., Zhang, Z., Bi, A., Xiang, H., He, X., Tian, X., Liu, W., Zhang, J., Peng, C., Zhu, Z., Huang, M., Li, Y., Zhuang, G., & Tan, L.
- Affiliations
- Chinese Academy of Sciences (CN)
- Shanghai Jiao Tong University (CN)
- Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 201210, China
- Corresponding author
- Li, Ying
- Zhuang, Guanglei
- Tan, Li
- ORCID
- Venue
- Journal of Medicinal Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2023-02-01
- Volume
- 66
- Issue
- 4
- Pages
- 2608-2621
- ISSN
- 0022-2623 (ISSN-L)
- 1520-4804
- OpenAlex
- W4318753744
- Cited by
- 18
- References
- 39
- Citations by year
- 2026: 4 · 2025: 6 · 2024: 5 · 2023: 3
- FWCI
- 1.84
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Journal of Medicinal Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Shanghai Municipal Education Commission (20161313)
- Science and Technology Commission of Shanghai Municipality (2019SHZDZX02)
- National Natural Science Foundation of China (21837004, 32170755, 81922047, 82151212, 82172596)
- Chinese Academy of Sciences (XDB39050500)
- Innovative Research Team of High-Level Local Universities in Shanghai (SHSMU-ZLCX20210200)
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis and biological activity
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Keywords
- PKM2, PKM2 inhibitors, allosteric inhibition, prodrugs, oral bioavailability, tumor growth inhibition, drug discovery
- MeSH terms
- Cysteine, Pyruvate Kinase, Drug Discovery
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