Flufenamic acid-based sulfonohydrazide and acetamide derivatives NSAI as inhibitors of multi-targets COX-1/COX-2/5-LOX: design, synthesis, in silico ADMET and binding mode studies

Although inflammation triggers immune-mediated healing and repair, chronic inflammation can result in several diseases. Cyclooxygenase (COX) enzymes are inhibited by NSAIDs, which are used to relieve the symptoms of inflammation. Meclofenamic and Zileuton are examples of dual COX/LOX inhibitors that provide improved stomach protection and safer cardiovascular characteristics. This study was aimed to develop anti-inflammatory medications with improved safety profiles by presenting novel flufenamate conjugates that combine 5-LOX inhibitor efficacy with COX inhibition. The COX-1 inhibition of compounds 14 and 15 was higher than that of Celecoxib (IC₅₀ = 77.4 µM), with an IC₅₀ range of 15-26 µM. Compounds 14 and 16 showed the best selectivity indices (ratio between IC₅₀ of COX-1 and COX-2), which were 5.01 and 5.86 µM, respectively. Conjugates 14 and 16 displayed excellent COX-2 inhibiting activity, with IC₅₀ values of 5.0 -17.6 µM. Outstanding 5-LOX inhibition was demonstrated by all conjugates, with IC₅₀ values varying between 0.6 and 8.5 µM. In RAW 264.7 cells, compounds 14 and 15 significantly decreased PGE2 levels to a range of 61-89 pg/mL in contrast to Celecoxib (119.9 pg/mL). Compounds 14 and 17 showed exceptional NO scavenging action, with IC₅₀ values of 0.238 × 10⁶ and 0.289 × 10⁶ µM, respectively. mTOR levels dramatically diminished for all conjugates. Conjugates 14 and 17 markedly raised levels nrf2. Molecular docking studies were used to validate the findings of this investigation.