Synthesis, Computational Analysis and In-vitro anti-inflammatory activities of Naproxen Derivatives by Lipoxygenase and Cyclooxygenase Inhibitory Pathway

Enzymes that facilitate the transformation of arachidonic acid into prostaglandins include Lipoxygenase and Cyclooxygenase, which play a crucial role in pro-inflammatory processes. Allopathic medications that inhibit cyclooxygenase are classified as Nonsteroidal anti-inflammatory drugs (NSAIDs), which exhibit significant anti-inflammatory properties, also have several adverse effects, particularly on the gastrointestinal mucosa, attributed to their various functional groups. Hence, present research work is designed to develop new, safer anti-inflammatory derivatives of naproxen that possess dual inhibition capabilities for both Cyclooxygenase and Lipoxygenase pathways, while also enhancing the safety profile.Four derivatives of Naproxen (NS9-NS12) were synthesized through a three-step process and were characterized by 1H NMR, 13CNMR, 19FNMR. Additionally, the safety profile was checked according to OECD guidelines 423 where a single dose of 2000 mg/kg (b.wt) of all derivatives was administered through oral route, while in- vitro anti-inflammatory activity was assessed through the Lipoxygenase and Cyclooxygenase inhibitory pathways. Derivatives were found safe up to dose 2000 mg/kg (b.wt) having no mortality and behavioral changes in any animals treated group. In Cyclooxygenase pathway, NS12 demonstrated high significant inhibitory effect against cyclooxygenase II, with values of 97.33±0.66, 95.78±0.38, 88.65±0.44, 84.67±0.28, 80.55±0.48,77.65±0.53 and 62.65±0.62 achieving an IC50 of <01 μM at concentration 1000-31.25 µg/mL compared with NS9, NS10 and NS11 on the same concentrations. In Lipoxygenase pathway, NS11 exhibited an IC50 of <01 μM, also within the concentration range of 1000-31.25 µg/mL, with percent inhibition values of 91.23± 0.48, 87.44± 0.80, 85.72± 0.61, 81.65± 0.72, 77.86± 0.38, 73.45± 0.77. In comparison with Cyclooxygenase and Lipoxygenase activities, NS12 displayed greater percent inhibition for Cyclooxygenase while for Lipoxygenase NS11. From the docking conformation compound NS11 was found good docking score of -15.9737 formed four polar interactions, one pi-cation and five pi-H linkages with the active residues of the lipoxygenase enzyme.