Myoglobin microplate assay to evaluate prevention of protein peroxidation
| Autor: | Sara S. Marques, Ana Mota, Tânia Soares, Luís M. Magalhães, Barbara Korsak, Salette Reis, Marcela A. Segundo |
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| Rok vydání: | 2015 |
| Předmět: |
Indoles
Hemeprotein Free Radicals Taurine Clinical Biochemistry Pharmaceutical Science Angiotensin-Converting Enzyme Inhibitors Oxidative phosphorylation Protein oxidation Antioxidants Analytical Chemistry Fatty Acids Monounsaturated chemistry.chemical_compound Enalapril Drug Discovery medicine Humans Dimethyl Sulfoxide Fluvastatin Spectroscopy chemistry.chemical_classification Reactive oxygen species Ethanol Autoxidation Myoglobin Chemistry Temperature Hydrogen Peroxide Glutathione Hydrogen-Ion Concentration Oxidants Peroxides Oxygen Kinetics Oxidative Stress Biochemistry Solvents Biological Assay Spectrophotometry Ultraviolet Hydroxymethylglutaryl-CoA Reductase Inhibitors Oxidation-Reduction medicine.drug |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 114:305-311 |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/j.jpba.2015.06.006 |
| Popis: | The current therapeutic strategies are based on the design of multifunctional drug candidates able to interact with various disease related targets. Drugs that have the ability to scavenge reactive oxygen species (ROS), beyond their main therapeutic action, may prevent the oxidative damage of biomolecules. Therefore, analytical approaches that monitor in a continuous mode the ability of drugs to counteract peroxidation of physiologically relevant biotargets are required. In the present work, a microplate spectrophotometric assay is proposed to evaluate the ability of selected cardiovascular drugs, including angiotensin-converting enzyme (ACE) inhibitors, β -blockers and statins to prevent protein peroxidation. Myoglobin, which is a heme protein, and peroxyl radicals generated from thermolysis of 2,2'-azo-bis(2-amidinopropane) dihydrochloride at 37 °C, pH 7.4 were selected as protein model and oxidative species, respectively. Myoglobin peroxidation was continuously monitored by the absorbance decrease at 409 nm and the ability of drugs to counteract protein oxidation was determined by the calculation of the area under the curve upon the myoglobin oxidation. Fluvastatin (AUC₅₀=12.5 ± 1.2 μM) and enalapril (AUC₅₀=15.2 ± 1.8 μM) showed high ability to prevent myoglobin peroxidation, providing even better efficiency than endogenous antioxidants such as reduced glutathione. Moreover, labetalol, enalapril and fluvastatin prevent the autoxidation of myoglobin, while glutathione showed a pro-oxidant effect. |
| Databáze: | OpenAIRE |
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