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Metabolic stability of the new antimalarial drug leads is determined using Human Liver Microsome (HLM) and specific cytochrome P450 enzyme (CYP2C8) taking the clinically used antimalarial drug chloroquine as a positive control. Experiment is done using standard methods. All the assays were conducted in 0.5 M phosphate buffer at pH 7.4. In general the metabolic reaction was initiated by adding 1 mM NADPH and 0.5 mg of enzyme. Incubations were done with time frequency of 0 hr, 1 hr, and 2 hrs at 37°C and the reactions were terminated by adding acetonitrile in the equal amounts of the assay mixture taken. The samples were centrifuged for 15 minutes at 10,000×g at 4°C and an aliquot of the supernatant was subjected to analysis using HPLC as well as LC-MS to confirm the masses of the drug and/or metabolite(s), if any. While chloroquine was found to be metabolized in a predictable manner by both HLM and CYP2C8, the drug leads were metabolically stable at similar experimental conditions. This study demonstrated that the new drug leads are worth conducting further preclinical evaluations.
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