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Synthesis and physicochemical properties of novel lophine derivatives as chemiluminescent in vitro activators for detection of free radicals

European Biophysics Journal volume 44, pages 623–634 (2015)Cite this article

Abstract

The overproduction of free radicals and reactive oxygen species (ROS) has been proved as a basic damage mechanism and cause for oxidative stress. Their measurement is often hindered by the low signal. This could be resolved with the application of luminescent probes (lophines, luminol, lucigenin, etc.). The focus of this study is to synthesize and describe the spectral properties and physicochemical characteristics of lophine and its derivatives as new chemiluminescent in vitro activators. The prepared luminophores are analogues of lophine. Their absorption maxima are in the range 329–340 nm, with good-to-high extinction coefficients. Their spectral properties are measured in methanol and buffer solutions with pH 3.5, 7.4 and 8.5. Same conditions were applied in the systems for chemiluminescent assay in vitro: (1) Fenton’s (Fe2++H2O2) for the generation of ·OH and –OH species, (2) Hydrogen peroxide (H2O2), (3) Iron (II) sulfate (FeSO4), (4) Glutathione-peroxidase, monitoring the deactivation of H2O2, (5) Ascorbic acid-Fenton’s reagent: Vit.C appears a strong oxidant, generating free-radical products when applied in higher than physiological concentrations, (6) Reduced α-nicotinamide adenine dinucleotide (NADH)-phenazine methosulfate—for the generation of superoxide radicals (O2 ·−). Lophine and all novel compounds do not alter the kinetics, except of the dimethyl amino substituted derivative (4-(3a,11b-dihydro-1H-imidazo[4,5-f][1,10]phenanthrolin-2-yl)-N,N-dimethylaniline) in the glutathione-peroxidase system, at pH 8.5. Same derivative showed a comparable or higher activity than Lucigenin and Rhodamine 6G. In neutral and acidic medium, in the Fenton’s system, Rhodamine 6G was the most appropriate probe. In alkaline pH and oxidant H2O2, Lucigenin induced a signal twice as strong as the signal compared to all other activators.

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Notes

  1. http://en.wikipedia.org/wiki/Rhodamine, http://chemicalland21.com/specialtychem/finechem/VICTORIA%20BLUE%20B.htm.

  2. http://en.wikipedia.org/wiki/Luminol.

  3. http://chemicalland21.com/specialtychem/finechem/VICTORIA%20BLUE%20B.htm.

  4. http://en.wikipedia.org/wiki/Lucigenin.

  5. http://sciencelinks.jp/j-east/article/200324/000020032403A0643390.php.

  6. http://journals.iucr.org/e/issues/2009/04/00/hb2915/hb2915bdy.html#BB16.

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Acknowledgments

This research was financially supported by Sofia University “St. Kliment Ohridski”, according to the project “Synthesis and study of the physical and chemical characteristics of lophine and its derivatives as novel chemiluminescent probes for medical and biological research”, No 2666—Scientific Research Centre. We are very thankful for the editorial support of Dr. Mihail Mitov—Research Associate and Michael Alstott—Laboratory Technician Senior at the Redox Metabolism Shared Resource Facility (RM SRF), Markey Cancer Center, University of Kentucky: 1095 V.A. Drive Health Sciences Research Building, room 227, Lexington, KY 40536-0305, Phone: (859) 323-1106.

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Authors and Affiliations

  1. Biophysics and Medical Physics, Faculty of Physics, Sofia University “St. Kliment Ohridski”, 5 James Boucher Blvd., 1164, Sofia, Bulgaria

    Elitsa Pavlova

  2. Department of Applied Organic Chemistry, Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Boucher Blvd., 1164, Sofia, Bulgaria

    Stefka Kaloyanova, Todor Deligeorgiev & Nedyalko Lesev

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  1. Elitsa Pavlova
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  2. Stefka Kaloyanova
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  3. Todor Deligeorgiev
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Correspondence to Elitsa Pavlova.

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Pavlova, E., Kaloyanova, S., Deligeorgiev, T. et al. Synthesis and physicochemical properties of novel lophine derivatives as chemiluminescent in vitro activators for detection of free radicals. Eur Biophys J 44, 623–634 (2015). https://doi.org/10.1007/s00249-015-1056-3

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Keywords

  • Lophine
  • ROS
  • Chemiluminescent probes