Abstract

Small ubiquitin-like modifier (SUMO) family proteins regulate target-protein functions by post-translational modification. However, a potent and selective inhibitor targeting the SUMO pathway has been lacking. Here we describe ML-792, a mechanism-based SUMO-activating enzyme (SAE) inhibitor with nanomolar potency in cellular assays. ML-792 selectively blocks SAE enzyme activity and total SUMOylation, thus decreasing cancer cell proliferation. Moreover, we found that induction of the MYC oncogene increased the ML-792-mediated viability effect in cancer cells, thus indicating a potential application of SAE inhibitors in treating MYC-amplified tumors. Using ML-792, we further explored the critical roles of SUMOylation in mitotic progression and chromosome segregation. Furthermore, expression of an SAE catalytic-subunit (UBA2) S95N M97T mutant rescued SUMOylation loss and the mitotic defect induced by ML-792, thus confirming the selectivity of ML-792. As a potent and selective SAE inhibitor, ML-792 provides rapid loss of endogenously SUMOylated proteins, thereby facilitating novel insights into SUMO biology.

  • Compound

    {(1R,2S,4R)-4-[(5-{[1-(3-Bromobenzyl)-1H-pyrazol-3-yl]carbonyl}pyrimidin-4-yl)amino]-2-hydroxycyclopentyl}methyl sulfamate

  • Compound

    1-(3-Bromobenzyl)-1H-pyrazole-3-carbaldehyde

  • Compound

    [1-(3-Bromobenzyl)-1H-pyrazol-3-yl](4-chloropyrimidin-5-yl)methanol

  • Compound

    [1-(3-Bromobenzyl)-1H-pyrazol-3-yl](4-chloropyrimidin-5-yl)methanone

  • Compound

    [1-(3-Bromobenzyl)-1H-pyrazol-3-yl][4-({(1R,3R,4S)-3-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4-[(triisopropylsilyl)oxy]cyclopentyl}amino)pyrimidin-5-yl]methanone

  • Compound

    [1-(3-Bromobenzyl)-1H-pyrazol-3-yl][4-({(1R,3R,4S)-3-(hydroxymethyl)-4-[(triisopropylsilyl)oxy]cyclopentyl}amino)pyrimidin-5-yl]methanone

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Acknowledgements

We sincerely thank J. Klekota for bioinformatics support and K. Galvin, P. Veiby, C. Lou, M. Manfredi and C. Claiborne for insightful comments. We thank R. Hay (University of Dundee) for p300 cells.

Author information

Author notes

    • Neil Bence

    Present address: Nurix, Inc., San Francisco, California, USA.

Affiliations

  1. Oncology Drug Discovery Unit, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts, USA.

    • Xingyue He
    • , Jessica Riceberg
    • , Teresa Soucy
    • , Erik Koenig
    • , James Minissale
    • , Melissa Gallery
    • , Hugues Bernard
    • , Xiaofeng Yang
    • , Hua Liao
    • , Claudia Rabino
    • , Pooja Shah
    • , Kristina Xega
    • , Zhong-hua Yan
    • , Mike Sintchak
    • , John Bradley
    • , He Xu
    • , Matt Duffey
    • , Dylan England
    • , Hirotake Mizutani
    • , Zhigen Hu
    • , Jianping Guo
    • , Ryan Chau
    • , Lawrence R Dick
    • , James E Brownell
    • , John Newcomb
    • , Steve Langston
    • , Eric S Lightcap
    • , Neil Bence
    •  & Sai M Pulukuri

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Contributions

X.H., E.S.L., N.B. and S.M.P. conceived and designed the experiments. X.H., J.R., M.G., H.B., C.R., K.X., Z.Y. and J.B. performed cell biology experiments. T.S. and J.M. performed in vitro enzymatic assays. X.Y. and H.L. performed mass spectrometry analysis. M.S. provided structural prediction. H.X., M.D., D.E., H.M., Z.H., J.G., R.C. and S.L. designed chemical compounds and/or performed chemical synthesis. E.K., P.S. and E.S.L. performed RNA-seq analysis. All authors discussed the data and provided scientific input. X.H., T.S., E.K., L.R.D., J.E.B., J.N., S.L., E.S.L., N.B. and S.M.P. wrote and/or edited the paper.

Competing interests

Takeda Pharmaceuticals International Co. provided complete financial support for the study design, data collection and analysis, decision to publish, preparation of the manuscript and salaries for authors X.H., J.R., T.S., E.K., J.M., M.G., H.B., X.Y., H.L., C.R., P.S., K.X., Z.Y., M.S., J.B., H.X., M.D., D.E., H.M., Z.H., J.G., R.C., L.R.D., J.E.B., J.N., S.L., E.S.L., N.B. and S.M.P. during the time when studies were conducted. Patent application WO2015/002994 has been filed for compound ML-792.

Corresponding authors

Correspondence to Xingyue He or Sai M Pulukuri.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Results, Supplementary Figures 1–22.

  2. 2.

    Supplementary Note 1

    Synthesis and Characterization of ML-792.

Excel files

  1. 1.

    Supplementary Table 1

    ML-792 activity against a panel of 366 ATP-utilizing enzymes.

  2. 2.

    Supplementary Table 2

    RNAseq analysis on HCT116 cells harboring UBA2 shRNA s or treated with ML-792.

  3. 3.

    Supplementary Table 3

    quantitative proteomic profiling in HCT116 cells treated with DMSO or ML-792.

Videos

  1. 1.

    phase-contrast time-lapse microscopy with DMSO treated HCT116 cells

  2. 2.

    phase-contrast time-lapse microscopy with ML-792 treated HCT116 cells

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DOI

https://doi.org/10.1038/nchembio.2463

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