Determination of Michaelis-Menten characteristics for defined substrates in experimental systems. Determination of binding characteristics of pharmaceutically active form in supersomal fraction of one human cytochrome P-450 form
Abstract
The contractual research included experimental in vitro characterization of enzymatic kinetics and binding affinity test for two pharmaceutically active substances and two medicinal drug forms in three relevant enzymatic systems. The three enzymatic preparations, two human microsomal fractions (male, female) and one isoform of human cytochrome P450 were employed for experimental approaches.
With tested substances was optimized experimental work-flow including incubations with enzymatic substrates, extraction protocol, analytical separation employing HPLC with UV-VIS or MS detection. The obtained values were used for quantitative and qualitative evaluation and determination of kinetics parameters.