In Vitro Enzyme Kinetics and NMR-Based Product Elucidation for Glutathione S-Conjugation of the Anticancer Unsymmetrical Bisacridine C-2028 in Liver Microsomes and Cytosol: Major Role of Glutathione S-Transferase M1-1 Isoenzyme

This work is the next step in studying interplay between C-2028 (anticancer-active unsymmetrical bisacridine developed our group) and glutathione S-transferase/glutathione (GST/GSH) system. Here, we analyzed concentration- pH-dependent GSH conjugation of rat liver microsomes cytosol. We also applied three recombinant human GST isoenzymes, which altered expression was found various tumors. The formation S-conjugate subfractions followed Michaelis-Menten kinetics. that conjugated with preferentially by GSTM1-1, revealing a sigmoidal kinetic model. Using colorimetric assay (MTT test), initially assessed cellular GST/GSH-dependent biotransformation relation to cytotoxicity against Du-145 prostate cancer cells presence or absence modulator biosynthesis. Pretreatment buthionine sulfoximine resulted decrease, suggesting possible GSH-mediated bioactivation process. Altogether, results confirmed importance metabolism, humans must consider when planning treatment strategy. Finally, nuclear magnetic resonance spectroscopy elucidated structure GSH-derived product C-2028. Hence, synthesizing compound standard necessary for further advanced biological bioanalytical investigations will be achievable.