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A new series of acetohydroxamates shows in vitro and in vivo anticancer activity against melanoma

Summary

Cancer treatment is challenging, mainly due to high levels of drug toxicity and the resistance of tumours to chemotherapy. Hydroxamic acid derivatives have recently aroused attention due to their potential to treat malignancies. In the present study, we sought to investigate the anticancer effects of a new series of synthetic acetohydroxamates. The in vitro cytotoxic and antiproliferative effects of 11 synthetic acetohydroxamates were evaluated against the melanoma cell line A375. Apoptosis, cell cycle, and autophagy assays were employed to elucidate the cell death pathways induced by the compounds. The in vivo pharmacokinetic profiles of the most promising compounds were determined in CD-1 mice, while the in vivo antitumour efficacies were evaluated using the A375 melanoma xenograft model in nude mice. MTT assays revealed that all compounds presented concentration-dependent cytotoxicity against the A375 cell line. AKS 61 produced the most favourable antiproliferative activity according to the sulphorhodamine B and clonogenic assays. AKS 61 treatment resulted in decreased mitochondrial membrane potential and increased apoptosis and autophagy in the A375 cell line. However, AKS 61 failed to prevent in vivo tumour growth in a melanoma xenograft, whereas compound AKS 7 was able to inhibit tumour growth when administered orally. These in vivo findings may be explained by a more favourable pharmacokinetic profile presented by AKS 7 when compared to AKS 61. Taken together, these results suggest that acetohydroxamates have potential anticancer effects and will guide future optimisation of these molecules to allow for further non-clinical development.

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Abbreviations

ATCC:

American Type Culture Collection

CMCiPS :

Cardiomyocytes derived from induced pluripotent stem cells

FDA:

Food and Drug Administration

GI50 :

Growth inhibition 50%

HDAC:

Histone deacetylase

MMP:

Mitochondrial membrane potential

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NHDF:

Non-tumoural human dermal fibroblasts

NHLF:

Non-tumoural human lung fibroblasts

SrB:

Sulphorhodamine B

TCA:

Trichloroacetic acid

TGI:

Total growth inhibition

UPLC-MS/MS:

Ultra-performance liquid chromatography-tandem mass spectrometry

References

  1. 1.

    Holohan C, Van Schaeybroeck S, Longley DB, Johnston PG (2013) Cancer drug resistance : an evolving paradigm. Nat Publ Group 13:714–726. https://doi.org/10.1038/nrc3599

  2. 2.

    Fulda S (2009) Tumor resistance to apoptosis. Int J Cancer 124:511–515. https://doi.org/10.1002/ijc.24064

  3. 3.

    Lefranc F, Sadeghi N, Camby I, Metens T, Dewitte O, Kiss R (2006) Present and potential future issues in glioblastoma treatment. Expert Rev Anticancer Ther 6:719–732. https://doi.org/10.1586/14737140.6.5.719

  4. 4.

    Mohammad RM, Muqbil I, Lowe L, Yedjou C, Hsu H-Y, Lin L-T, Siegelin MD, Fimognari C, Kumar NB, Dou QP, Yang H, Samadi AK, Russo GL, Spagnuolo C, Ray SK, Chakrabarti M, Morre JD, Coley HM, Honoki K, Fujii H, Georgakilas AG, Amedei A, Niccolai E, Amin A, Ashraf SS, Helferich WG, Yang X, Boosani CS, Guha G, Bhakta D, Ciriolo MR, Aquilano K, Chen S, Mohammed SI, Keith WN, Bilsland A, Halicka D, Nowsheen S, Azmi AS (2015) Broad targeting of resistance to apoptosis in cancer. Semin Cancer Biol 35:S78–S103. https://doi.org/10.1016/j.semcancer.2015.03.001

  5. 5.

    Huguet F, Melet A, Alves de Sousa R, Lieutaud A, Chevalier J, Maigre L, Deschamps P, Tomas A, Leulliot N, Pages JM, Artaud I (2012) Hydroxamic acids as potent inhibitors of Fe II and Mn II E. coli methionine aminopeptidase: biological activities and X-ray structures of oxazole hydroxamate-EcMetAP-Mn complexes. ChemMedChem 7:1020–1030. https://doi.org/10.1002/cmdc.201200076

  6. 6.

    Day JA, Cohen SM (2013) Investigating the selectivity of metalloenzyme inhibitors. J Med Chem 56:7997–8007. https://doi.org/10.1021/jm401053m

  7. 7.

    Gupta SP, Sharma A (2013) The chemistry of Hydroxamic acids. In: Gupta SP (ed) Hydroxamic acids: a unique family of chemicals with multiple biological activities. Springer, Berlin, p 311

  8. 8.

    Curtin M, Glaser K (2003) Histone deacetylase inhibitors: the Abbott experience. Curr Med Chem 10:2373–2392. https://doi.org/10.1097/CAD.0000000000000040

  9. 9.

    KrennHrubec K, Marshall BL, Hedglin M, Verdin E, Ulrich SM (2007) Design and evaluation of ‘Linkerless’ hydroxamic acids as selective HDAC8 inhibitors. Bioorg Med Chem Lett 17:2874–2878. https://doi.org/10.1016/j.bmcl.2007.02.064

  10. 10.

    Kaliszczak M, Trousil S, Åberg O, Perumal M, Nguyen Q-D, Aboagye EO (2013) A novel small molecule hydroxamate preferentially inhibits HDAC6 activity and tumour growth. Br J Cancer 108:342–350. https://doi.org/10.1038/bjc.2012.576

  11. 11.

    Minjie S, Defei H, Zhimin H, Weiding W, Yuhua Z (2015) Targeting pancreatic cancer cells by a novel hydroxamate-based histone deacetylase (HDAC) inhibitor ST-3595. Tumor Biol 36:9015–9022. https://doi.org/10.1007/s13277-015-3537-5

  12. 12.

    Manal M, Chandrasekar MJN, Gomathi Priya J, Nanjan MJ (2016) Inhibitors of histone deacetylase as antitumor agents: a critical review. Bioorg Chem 67:18–42. https://doi.org/10.1016/j.bioorg.2016.05.005

  13. 13.

    Aksenov AV, Smirnov AN, Magedov IV, Reisenauer MR, Aksenov NA, Aksenova IV, Pendleton AL, Nguyen G, Johnston RK, Rubin M, De Carvalho A, Kiss R, Mathieu V, Lefranc F, Correa J, Cavazos DA, Brenner AJ, Bryan BA, Rogelj S, Kornienko A, Frolova LV (2015) Activity of 2-aryl-2-(3-indolyl)acetohydroxamates against drug-resistant cancer cells. J Med Chem 58:2206–2220. https://doi.org/10.1021/jm501518y

  14. 14.

    Vichai V, Kirtikara K (2006) Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc 1:1112–1116. https://doi.org/10.1038/nprot.2006.179

  15. 15.

    Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG, Simera I et al (2010) Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol 8:e1000412. https://doi.org/10.1371/journal.pbio.1000412

  16. 16.

    Jung J (2014) Human tumor xenograft models for preclinical assessment of anticancer drug development. Toxicol Res 30:1–5. https://doi.org/10.5487/TR.2014.30.1.001

  17. 17.

    Tessier P, Smil DV, Wahhab A, Leit S, Rahil J, Li Z, Déziel R, Besterman JM (2009) Diphenylmethylene hydroxamic acids as selective class IIa histone deacetylase inhibitors. Bioorg Med Chem Lett 19:5684–5688. https://doi.org/10.1016/j.bmcl.2009.08.010

  18. 18.

    Al-Qatati A, Aliwaini S (2017) Combined pitavastatin and dacarbazine treatment activates apoptosis and autophagy resulting in synergistic cytotoxicity in melanoma cells. Oncol Lett 14:7993–7999. https://doi.org/10.3892/ol.2017.7189

  19. 19.

    Brown JM, Attardi LD (2005) The role of apoptosis in cancer development and treatment response. Nat Rev Cancer 5:231–237. https://doi.org/10.1038/nrc1560

  20. 20.

    Todaro M, Lombardo Y, Francipane MG, Alea MP, Cammareri P, Iovino F, Di Stefano AB, Di Bernardo C, Agrusa A, Condorelli G, Walczak H, Stassi G (2008) Apoptosis resistance in epithelial tumors is mediated by tumor-cell-derived interleukin-4. Cell Death Differ 15:762–772. https://doi.org/10.1038/sj.cdd.4402305

  21. 21.

    Eisenberg-Lerner A, Bialik S, Simon H-U, Kimchi A (2009) Life and death partners: apoptosis, autophagy and the cross-talk between them. Cell Death Differ 16:966–975. https://doi.org/10.1038/cdd.2009.33

  22. 22.

    Lee T-G, Jeong E-H, Kim SY, Kim H-R, Kim CH (2015) The combination of irreversible EGFR TKIs and SAHA induces apoptosis and autophagy-mediated cell death to overcome acquired resistance in EGFRT790M-mutated lung cancer. Int J Cancer 136:2717–2729. https://doi.org/10.1002/ijc.29320

  23. 23.

    Soares AS, Costa VM, Diniz C, Fresco P (2014) Combination of cl-IB-MECA with paclitaxel is a highly effective cytotoxic therapy causing mTOR-dependent autophagy and mitotic catastrophe on human melanoma cells. J Cancer Res Clin Oncol 140:921–935. https://doi.org/10.1007/s00432-014-1645-z

  24. 24.

    Walker DK, Jones RM, Nedderman ANR, Wright PA (2010) Primary, secondary and tertiary amines and their Isosteres. In: Smith DA (ed) Metabolism, pharmacokinetics, and toxicity of functional groups: impact of chemical building blocks on ADMET, 1st edn. RSC Publishing, Cambridge, p 510

  25. 25.

    Bhattacharyya GS (2010) Oral systemic therapy: Not all “win-win.”. India J Med Paediatr Oncol 31:1–3. https://doi.org/10.4103/0971-5851.68844

  26. 26.

    Lipinski CA (2004) Lead- and drug-like compounds: the rule-of-five revolution. Drug Discov Today Technol 1:337–341. https://doi.org/10.1016/j.ddtec.2004.11.007

  27. 27.

    Schwab M, Schaeffeler E, Brauch H (2012) Anticancer drugs. In: Anzenbacher P, Zanger UM (eds) Metabolism of drugs and other xenobiotics. Wiley-VCH, Weinheim, p 753

  28. 28.

    Clive S, Woo MM, Nydam T, Kelly L, Squier M, Kagan M (2012) Characterizing the disposition, metabolism, and excretion of an orally active pan-deacetylase inhibitor, panobinostat, via trace radiolabeled 14C material in advanced cancer patients. Cancer Chemother Pharmacol 70:513–522. https://doi.org/10.1007/s00280-012-1940-9

  29. 29.

    Calvo E, Reddy G, Boni V, García-Cañamaque L, Song T, Tjornelund J, Choi MR, Allen LF (2016) Pharmacokinetics, metabolism, and excretion of 14C-labeled belinostat in patients with recurrent or progressive malignancies. Investig New Drugs 34:193–201. https://doi.org/10.1007/s10637-015-0321-8

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Acknowledgements

This work was supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Instituto Nacional de Ciência e Tecnologia (INCT-INOVAMED), Fundação de Apoio à Pesquisa de Santa Catarina (FAPESC), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The authors gratefully thank Edir Rezende and Ana Caroline Luz Machado for their technical support. Financial support from the Russian Science Foundation (Grant #18-13-00238) is gratefully acknowledged.

Funding

This work was supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Instituto Nacional de Ciência e Tecnologia (INCT-INOVAMED), Fundação de Apoio à Pesquisa de Santa Catarina (FAPESC), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Financial support from the Russian Science Foundation (Grant #18–13-00238) is gratefully acknowledged.

Author information

G.C.S., C.G.M., E.C.S., M.H., R.C.S., A.K., R.M. and J.B.C. contributed to the study design. G.C.S., C.G.M., E.C.S., M.H., R.C.S. and R.M. conducted experiments. G.C.S., C.G.M., E.C.S., M.H. and R.C.S. performed the analysis of the data. A.V.A, N.A.A, D.A.A and A.K. contributed to the design and synthesis of the compounds. G.C.S., C.G.M., E.C.S., R.C.S., A.K., R.M. and J.B.C. contributed to the writing of the manuscript.

Correspondence to João B. Calixto.

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Conflict of interest

Gabriela C Segat declares that she has no conflict of interest. Camila G Moreira declares that she has no conflict of interest. Evelyn C Santos declares that she has no conflict of interest. Melina Heller declares that she has no conflict of interest. Raquel C Schwanke declares that she has no conflict of interest. Alexander V Aksenov declares that he has no conflict of interest. Nicolai A Aksenov declares that he has no conflict of interest. Dmitrii A Aksenov declares that he has no conflict of interest. Alexander Kornienko declares that he has no conflict of interest. Rodrigo Marcon declares that he has no conflict of interest. João B Calixto declares that he has no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Segat, G.C., Moreira, C.G., Santos, E.C. et al. A new series of acetohydroxamates shows in vitro and in vivo anticancer activity against melanoma. Invest New Drugs (2019). https://doi.org/10.1007/s10637-019-00849-6

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Keywords

  • Acetohydroxamate
  • Drug development
  • Cancer
  • Melanoma