Back to Search
Start Over
Myoglobin microplate assay to evaluate prevention of protein peroxidation.
- Source :
-
Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2015 Oct 10; Vol. 114, pp. 305-11. Date of Electronic Publication: 2015 Jun 10. - Publication Year :
- 2015
-
Abstract
- 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.<br /> (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Subjects :
- Angiotensin-Converting Enzyme Inhibitors chemistry
Biological Assay
Dimethyl Sulfoxide chemistry
Enalapril chemistry
Ethanol chemistry
Fatty Acids, Monounsaturated chemistry
Fluvastatin
Free Radicals chemistry
Glutathione chemistry
Humans
Hydrogen Peroxide chemistry
Hydrogen-Ion Concentration
Hydroxymethylglutaryl-CoA Reductase Inhibitors chemistry
Indoles chemistry
Kinetics
Oxidants chemistry
Oxidation-Reduction
Oxidative Stress
Oxygen chemistry
Solvents chemistry
Spectrophotometry, Ultraviolet
Taurine chemistry
Temperature
Antioxidants chemistry
Myoglobin chemistry
Peroxides chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1873-264X
- Volume :
- 114
- Database :
- MEDLINE
- Journal :
- Journal of pharmaceutical and biomedical analysis
- Publication Type :
- Academic Journal
- Accession number :
- 26093510
- Full Text :
- https://doi.org/10.1016/j.jpba.2015.06.006