5-Lipoxygenase (5-LOX) Inhibition Assay

The 5-LOX potentials of the compounds (4, 9–11) was determined by the previously established reported method.2 Different dilutions (500–31.25 μg/mL) of the synthesized compound were prepared. The lipoxygenase enzyme solution was also prepared 10,000 U/mL. The substrate linoleic acid of 80 mM was also prepared. The phosphate buffer solution of 50 mM having pH 6.3 was also prepared. A mixture of all the solutions (2 mL) having equivalent volumes of all were prepared and mixed thoroughly. The dilutions made of the compounds and also that of the standard drug were also added to the mixture. The rate of reaction was found for the compounds and control. The enzyme activity was measured at 234 nm by using a double beam spectrophotometer. The percentage activity/inhibition was determined by raising the absorbance of the compounds compared to the negative control. The zileuton was used as a positive control.