Myoglobin microplate assay to evaluate prevention of protein peroxidation

Publication date: 10 October 2015 Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 114 Author(s): Sara S. Marques , Luís M. Magalhães , Ana I.P. Mota , Tânia R.P. Soares , Barbara Korsak , Salette Reis , Marcela A. Segundo 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 409nm 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±1.2μM) and enalapril (AUC50 =15.2±1.8μM) ...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research