Prism 8 analysis

Statistical analyses were performed using GraphPad Prism 8 analysis (GraphPad Software, Inc., San Diego, CA, USA). Significance differences between groups were detected using one-way ANOVA at p < 0.05.

This assay examined the inhibitory potency of the target structures towardboth COX isoforms via colorimetric screening assay kit (Cayman Chemical, MI, United States) based on the directions provided by the manufacturer. ELISA reader Thermo Scientific Multiskan® EX (Thermo scientific, United States) was used. IC₅₀ (concentration at which there was 50% inhibition) values were calculated using GraphPad Prism 8 analysis software (Graph-Pad, San Diego, CA, United States) to form the dose–response curves of eight concentrations of each test compound. Serial dilutions of CLX, as a reference standard, and tested compounds at concentrations of 1000, 300, 100, 30, 10, 3, 1, and 0.30 nM were used in the assay. IC₅₀ of the tested compounds were determined using three different experiments. The selectivity (SI) of each ompound was measured using the COX-2/COX-1 ratio, which compares the IC₅₀ of COX-2 and COX-1.