Abstract
Oximes, investigated as antidotes against organophosphates (OP) poisoning, are known to display toxic effects on a cellular level, which could be explained beyond action on acetylcholinesterase as their main target. To investigate this further, we performed an in vitro cell-based evaluation of effects of two structurally diverse oxime groups at concentrations of up to 800 μM, on several cell models: skeletal muscle, kidney, liver, and neural cells. As indicated by our results, compounds with an imidazolium core induced necrosis, unregulated cell death characterized by a cell burst, increased formation of reactive oxygen species, and activation of antioxidant scavenging. On the other hand, oximes with a pyridinium core activated apoptosis through specific caspases 3, 8, and/or 9. Interestingly, some of the compounds exhibited a synergistic effect. Moreover, we generated a pharmacophore model for each oxime series and identified ligands from public databases that map to generated pharmacophores. Several interesting hits were obtained including chemotherapeutics and specific inhibitors. We were able to define the possible structural features of tested oximes triggering toxic effects: chlorine atoms in combination with but-2(E)-en-1,4-diyl linker and adding a second benzene ring with substituents such as chlorine and/or methyl on the imidazolium core. Such oximes could not be used in further OP antidote development research, but could be introduced in other research studies on new specific targets. This could undoubtedly result in an overall improved wider use of unexplored oxime database created so far in OP antidotes field of research in a completely new perspective.
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Data availability
All raw data are available by the corresponding authors upon reasonable request, and data are kept according to Croatian and Slovenian archives keeping law.
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Acknowledgements
We are grateful to Makso Herman Institute for medical Research and Occupational Health, Zagreb, Croatia, for language editing. We also thank Prof Kamil Kuca, Prof Kamil Musilek from the Faculty of Science, University of Hradec Kralove, Czech Republic and Prof Ines Primožič from the Faculty of Science, University of Zagreb, Croatia, for providing oximes for the experiments. We thank Prof Nino Sinčić and Dr Bojana Žegura for facilitating certain measurements at the School of Medicine, University of Zagreb, Croatia and National Institute of Biology, Ljubljana, Slovenia, respectively. We thank Dr Zrinka Kovarik, Institute for medical Research and Occupational Health, Zagreb, Croatia for the use of Biovia Discovery Studio Client v18.1 software (Dassault Systèmes, Vélizy-Villacoublay, France) provided through Croatian Science Foundation projects (grant numbers HrZZ-IP-2013-11-4307 and HrZZ-IP-2018-01-7683 to Z. Kovarik).
Funding
This study was supported by the Croatian Science Foundation (UIP-2017-05-7260 to M.K.), Slovenian Research Agency (J3-9263 to S.P. and J3-2523, P3-0043 and J7-8276 to S.P. and K.M.), Croatian–Slovenian Bilateral grant 2020–2021 (BI-HR/20-21-041) and Foundation of the Croatian Academy of Sciences and Arts (10-102/414-254-2018 to A.Z.).
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Conceptualization: MK and AZ; experiment performing: AZ, NM, PM, JM, KM; formal analysis and investigation: MK, SP, JP; writing—original draft preparation: MK and AZ; writing—review and editing: MK, AZ, NM, PM, JM, KM, JP, SP; funding acquisition: MK, AZ, SP.
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All studies on the primary human skeletal muscle cells reported here were conducted at the Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia. The use of human skeletal muscle cells (HSMC) was approved by the Republic of Slovenia Medical Ethics Committee (#71/05/12 and #0120-698/2017/4). The manuscript does not contain clinical studies or patient data.
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Zandona, A., Maraković, N., Mišetić, P. et al. Activation of (un)regulated cell death as a new perspective for bispyridinium and imidazolium oximes. Arch Toxicol 95, 2737–2754 (2021). https://doi.org/10.1007/s00204-021-03098-w
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DOI: https://doi.org/10.1007/s00204-021-03098-w