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3JC48-3 (methyl 4′-methyl-5-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-[1,1′-biphenyl]-3-carboxylate): a novel MYC/MAX dimerization inhibitor reduces prostate cancer growth

Cancer Gene Therapy volume 29, pages 1550–1557 (2022)Cite this article

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Abstract

The proto-oncogene cellular myelocytomatosis (c-Myc) is a transcription factor that is upregulated in several human cancers. Therapeutic targeting of c-Myc remains a challenge because of a disordered protein tertiary structure. The basic helical structure and zipper protein of c-Myc forms an obligate heterodimer with its partner MYC-associated factor X (MAX) to function as a transcription factor. An attractive strategy is to inhibit MYC/MAX dimerization to decrease c-Myc transcriptional function. Several methods have been described to inhibit MYC/MAX dimerization including small molecular inhibitors and proteomimetics. We studied the effect of a second-generation small molecular inhibitor 3JC48-3 on prostate cancer growth and viability. In our experimental studies, we found 3JC48-3 decreases prostate cancer cells’ growth and viability in a dose-dependent fashion in vitro. We confirmed inhibition of MYC/MAX dimerization by 3JC48-3 using immunoprecipitation experiments. We have previously shown that the MYC/MAX heterodimer is a transcriptional repressor of a novel kinase protein kinase D1 (PrKD1). Treatment with 3JC48-3 upregulated PrKD1 expression and phosphorylation of known PrKD1 substrates: the threonine 120 (Thr-120) residue in beta-catenin and the serine 216 (Ser-216) in Cell Division Cycle 25 (CDC25C). The mining of gene expression in human metastatic prostate cancer samples demonstrated an inverse correlation between PrKD1 and c-Myc expression. Normal mice and mice with patient-derived prostate cancer xenografts (PDX) tolerated intraperitoneal injections of 3JC48-3 up to 100 mg/kg body weight without dose-limiting toxicity. Preliminary results in these PDX mouse models suggest that 3JC48-3 may be effective in decreasing the rate of tumor growth. In conclusion, our study demonstrates that 3JC48-3 is a potent MYC/MAX heterodimerization inhibitor that decreases prostate cancer growth and viability associated with upregulation of PrKD1 expression and kinase activity.

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Fig. 1: Gene and protein expression in prostate cancer cell lines.
Fig. 2: Cell viability assay and cell-cycle analysis.
Fig. 3: Protein expression, immunoprecipitation assay in PC-3 cells, cell viability assay in C4-2 PrKD1 cells following 3JC48-3 treatment, and gene expression analysis in metastatic human prostate cancer tissue.
Fig. 4: Maximum tolerated dose experiment by intraperitoneal injection for 3JC48-3 inhibitor in mice models at 50 mg/kg and 100 mg/kg.
Fig. 5: High-performance liquid chromatography (HPLC) analysis for detection and measurement of concentration of 3JC48-3 in serum from mice.
Fig. 6: 3JC48-3 drug tolerability studies.

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Data availability

The data used to generate Fig. 3D can be found at: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE8511. All other data generated and analyzed during this study can be found within this article.

References

  1. Zeng W, Sun H, Meng F, Liu Z, Xiong J, Zhou S, et al. Nuclear C-MYC expression level is associated with disease progression and potentially predictive of two year overall survival in prostate cancer. Int J Clin Exp Pathol. 2015;8:1878–88.

    PubMed  PubMed Central  Google Scholar 

  2. Yu C, Niu X, Jin F, Liu Z, Jin C, Lai L. Structure-based Inhibitor Design for the Intrinsically Disordered Protein c-Myc. Sci Rep. 2016;6:22298.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Grandori C, Cowley SM, James LP, Eisenman RN. The Myc/Max/Mad network and the transcriptional control of cell behavior. Annu Rev Cell Dev Biol. 2000;16:653–99.

    Article  CAS  PubMed  Google Scholar 

  4. Singh SS, Jois SD. Homo- and heterodimerization of proteins in cell signaling: inhibition and drug design. Adv Protein Chem Struct Biol. 2018;111:1–59.

    Article  CAS  PubMed  Google Scholar 

  5. Zhong L, Li Y, Xiong L, Wang W, Wu M, Yuan T, et al. Small molecules in targeted cancer therapy: advances, challenges, and future perspectives. Signal Transduct Target Ther. 2021;6:201.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Shin WH, Kumazawa K, Imai K, Hirokawa T, Kihara D. Current challenges and opportunities in designing protein-protein interaction targeted drugs. Adv Appl Bioinform Chem. 2020;13:11–25.

    PubMed  PubMed Central  Google Scholar 

  7. Yap JL, Wang H, Hu A, Chauhan J, Jung KY, Gharavi RB, et al. Pharmacophore identification of c-Myc inhibitor 10074-G5. Bioorg Med Chem Lett. 2013;23:370–4.

    Article  CAS  PubMed  Google Scholar 

  8. Chauhan J, Wang H, Yap JL, Sabato PE, Hu A, Prochownik EV, et al. Discovery of methyl 4′-methyl-5-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-[1,1′-biphenyl]-3-carboxylate, an improved small-molecule inhibitor of c-Myc-max dimerization.Chem Med Chem. 2014;9:2274–2285.

  9. Smith K, Dalton S. Myc transcription factors: key regulators behind establishment and maintenance of pluripotency. Regen Med. 2010;5:947–59.

    Article  CAS  PubMed  Google Scholar 

  10. Nickkholgh B, Sittadjody S, Rothberg MB, Fang X, Li K, Chou JW, et al. Beta-catenin represses protein kinase D1 gene expression by non-canonical pathway through MYC/MAX transcription complex in prostate cancer. Oncotarget 2017;8:78811–24.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Youssef I, Ricort JM. Deciphering the role of protein kinase D1 (PKD1) in cellular proliferation. Mol Cancer Res. 2019;17:1961–74.

    Article  CAS  PubMed  Google Scholar 

  12. Steinberg SF. Regulation of protein kinase D1 activity. Mol Pharmacol. 2012;81:284–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Sundram V, Chauhan SC, Jaggi M. Emerging roles of protein kinase D1 in cancer. Mol Cancer Res. 2011;9:985–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Du C, Jaggi M, Zhang C, Balaji KC. Protein kinase D1-mediated phosphorylation and subcellular localization of beta-catenin. Cancer Res. 2009;69:1117–24.

    Article  CAS  PubMed  Google Scholar 

  15. Nickkholgh B, Sittadjody S, Ordonez K, Rothberg MB, Balaji KC. Protein kinase D1 induces G1-phase cell-cycle arrest independent of Checkpoint kinases by phosphorylating Cell Division Cycle Phosphatase 25. Prostate 2019;79:1053–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Arsura M, Deshpande A, Hann SR, Sonenshein GE. Variant Max protein, derived by alternative splicing, associates with c-Myc in vivo and inhibits transactivation. Mol Cell Biol. 1995;15:6702–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Giffney HE, Cummins EP, Murphy EP, Brayden DJ, Crean D. Protein kinase D, ubiquitin and proteasome pathways are involved in adenosine receptor-stimulated NR4A expression in myeloid cells. Biochem Biophys Res Commun. 2021;555:19–25.

    Article  CAS  PubMed  Google Scholar 

  18. Wang S, Wang J, Lv X. Selection of reference genes for expression analysis in mouse models of acute alcoholic liver injury. Int J Mol Med. 2018;41:3527–36.

    CAS  PubMed  Google Scholar 

  19. Barrett T, Wilhite SE, Ledoux P, Evangelista C, Kim IF, Tomashevsky M, et al. NCBI GEO: archive for functional genomics data sets-update. Nucleic Acids Res. 2013;41:D991–5.

    Article  CAS  PubMed  Google Scholar 

  20. Edgar R, Domrachev M, Lash AE. Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res. 2002;30:207–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Chen C, Lin W, Huang Y, Chen X, Wang H, Teng L. The essential factors of establishing patient-derived tumor model. J Cancer. 2021;12:28–37.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Karantanos T, Corn PG, Thompson TC. Prostate cancer progression after androgen deprivation therapy: mechanisms of castrate resistance and novel therapeutic approaches. Oncogene 2013;32:5501–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Kiessling A, Sperl B, Hollis A, Eick D, Berg T. Selective inhibition of c-Myc/Max dimerization and DNA binding by small molecules. Chem Biol. 2006;13:745–51.

    Article  CAS  PubMed  Google Scholar 

  24. Jubb H, Blundell TL, Ascher DB. Flexibility and small pockets at protein-protein interfaces: new insights into druggability. Prog Biophys Mol Biol. 2015;119:2–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Lu H, Zhou Q, He J, Jiang Z, Peng C, Tong R, et al. Recent advances in the development of protein–protein interactions modulators: mechanisms and clinical trials. Signal Transduct Target Ther. 2020;5:213.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Berg T, Cohen SB, Desharnais J, Sonderegger C, Maslyar DJ, Goldberg J, et al. Small-molecule antagonists of Myc/Max dimerization inhibit Myc-induced transformation of chicken embryo fibroblasts. Proc Natl Acad Sci USA. 2002;99:3830.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Bedard PL, Hyman DM, Davids MS, Siu LL. Small molecules, big impact: 20 years of targeted therapy in oncology. Lancet 2020;395:1078–88.

    Article  CAS  PubMed  Google Scholar 

  28. Mansoori B, Mohammadi A, Davudian S, Shirjang S, Baradaran B. The different mechanisms of cancer drug resistance: a brief review. Adv Pharm Bull. 2017;7:339–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Lv D, Chen H, Feng Y, Cui B, Kang Y, Zhang P, et al. Small-molecule inhibitor targeting protein kinase D: a potential therapeutic strategy. Front Oncol. 2021;11:680221.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Madden SK, de Araujo AD, Gerhardt M, Fairlie DP, Mason JM. Taking the Myc out of cancer: toward therapeutic strategies to directly inhibit c-Myc. Mol Cancer. 2021;20:3.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This study was supported by the Department of Defense under Award Number W81XWH-19-1-0745 to K.C.B.

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Authors and Affiliations

  1. University of Florida, Department of Urology, Jacksonville, FL, 32209, USA

    Sanjeev Shukla, Victor Chalfant, Carlos Riveros, Teruko Osumi & K. C. Balaji

  2. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD, 21201, USA

    Steven Fletcher & Jay Chauhan

  3. Department of Radiation Oncology, Mayo Clinic Florida, Jacksonville, FL, 32224, USA

    Yuri Mackeyev, Pankaj Kumar Singh & Sunil Krishnan

Authors
  1. Sanjeev Shukla
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Contributions

Conceptualization by S.S., S.F., J.C., T.O., and K.C.B.; Methodology by S.S., S.F., J.C., T.O., and K.C.B.; Validation by S.F., S.K., and K.C.B.; Formal analysis by S.S., J.C., Y.M., P.K.S., and C.R.; Resources by S.F., S.K., and K.C.B.; Data curation by S.S., S.F., Y.M., P.K.S., and K.C.B.; Writing by S.S., V.C., and C.R.; Original draft by S.S., S.F., V.C., and C.R.; Writing – Review and Editing by S.S., V.C., C.R., and K.C.B.; Supervision by S.F., S.K., and K.C.B.; Project administration by K.C.B.

Corresponding author

Correspondence to K. C. Balaji.

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The authors declare no competing interests.

Ethical approval

This study was approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Florida. All procedures adhered with the most recent specifications of the National Research Council (US) Committee for the Care and Use of Laboratory Animals (2011).

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Shukla, S., Fletcher, S., Chauhan, J. et al. 3JC48-3 (methyl 4′-methyl-5-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-[1,1′-biphenyl]-3-carboxylate): a novel MYC/MAX dimerization inhibitor reduces prostate cancer growth. Cancer Gene Ther 29, 1550–1557 (2022). https://doi.org/10.1038/s41417-022-00455-4

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