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Pro-fluorescent probe with morpholine moiety and its reactivity towards selected biological oxidants |
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| Authors: | Julia Modrzejewska Aleksandra Grzelakowska Marcin Szala Radosław Michalski Małgorzata Zakłos-Szyda Radosław Podsiadły |
| Affiliation: | 1. Institute of Polymer and Dye Technology, Faculty of Chemistry, Lodz University of Technology, Stefanowskiego 16, Lodz, Poland Contribution: Data curation, Investigation, Writing - original draft;2. Institute of Polymer and Dye Technology, Faculty of Chemistry, Lodz University of Technology, Stefanowskiego 16, Lodz, Poland Contribution: Conceptualization, Formal analysis, Methodology;3. Institute of Polymer and Dye Technology, Faculty of Chemistry, Lodz University of Technology, Stefanowskiego 16, Lodz, Poland Contribution: Conceptualization, Methodology;4. Institute of Applied Radiation Chemistry, Faculty of Chemistry, Lodz University of Technology, Żeromskiego 116, Lodz, Poland Contribution: Conceptualization, Investigation, Methodology;5. Institute of Molecular and Industrial Biotechnology, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, Stefanowskiego 2/22, Lodz, Poland Contribution: Data curation, Formal analysis, Investigation, Methodology;6. Institute of Polymer and Dye Technology, Faculty of Chemistry, Lodz University of Technology, Stefanowskiego 16, Lodz, Poland |
| Abstract: | Biological oxidants participate in many processes in the human body. Their excessive production causes organelle damage, which may result in the accumulation of cytotoxic mediators and cell degradation and may manifest itself in various diseases. Peroxynitrite (ONOO−), hypochlorous acid (HOCl), hydrogen peroxide (H2O2), and peroxymonocarbonate (HOOCO2−) are important oxidants in biology, toxicology, and various pathologies. Derivatives of coumarin, containing an oxidant-sensitive boronate group, have been recently developed for the fluorescent detection of inflammatory oxidants. Here, we report the synthesis and characterization of 4-[2-(morpholin-4-yl)-2-oxoethyl]-2-oxo-2H-chromen-7-yl boronic acid ( MpC-BA ) as a fluorescent probe for the detection of oxidants, with better solubility in water, high stability and fast response time toward peroxynitrite and hypochlorous acid. The effectiveness of the MpC-BA probe for the detection of peroxynitrite was measured by adding bolus ONOO− or using the co-generating superoxide and nitrogen oxide system. MpC-BA is oxidized by ONOO− to 7-hydroxy-4-[2-(morpholin-4-yl)-2-oxoethyl]-2H-chromen-2-one ( MpC-OH ). However, peroxynitrite-specific product ( MpC-H ) is formed in the minor reaction pathway. MpC-OH is also yielded in the reaction of MpC-BA with HOCl, and the subsequent formation of a chlorinated MpC-OH gives a specific product for HOCl ( MpC-OHCl ). H2O2 slowly oxidizes MpC-BA . However, the addition of NaHCO3 increased the MpC-OH formation rate. We conclude that MpC-BA is potentially an improved fluorescent probe detecting peroxynitrite and hypochlorite in biological settings. Complementation of the fluorescence measurements by HPLC-based identification of chlorinated and reduced coumarin(s) will help identify the oxidants detected. |
| Keywords: | coumarin-based probes oxidant-sensitive probe peroxynitrite-specific product, HOCl-specific product, HPLC analysis redox probe |