An operationally simple, one-pot, convenient synthesis, and in vitro anti-inflammatory activity of some new spirotriazolotriazine derivatives

The wide biological actions of triazolotriazine hybrids, benzene-sulfonamide derivatives, exhibit a good chance of displaying in vitro anti-inflammatory effects. An operationally simple one-pot, three-component, and convenient synthesis method is used toward the synthesis of a series of diverse 2-(2-(4-amino)-1,3,5-triazaspiro-1,4-dien-2-yl) hydrazinyl)-N-(4-sulfamoylphenyl)-2-thioxoacetamides, which generated via reaction of 2-hydrazinyl-N-(4-sulfamoylphenyl)-2-thioxoacetamide, cyanoguanidine, and cyclic/acyclic ketones and were characterized by spectral analysis (IR, Nuclear Magnetic Resonance: 1H-NMR, 13C-NMR, and elemental analysis). The spirotriazolotriazine derivatives are obtained with up to 95% yield, and their structure-activity relationship was discussed. All compounds were tested for in vitro anti-inflammatory potential using the inhibition of RBC hemolysis technique and COX inhibition assay. The results from RBC hemolysis technique demonstrated that almost all evaluated compounds exhibited significant in vitro anti-inflammatory efficacy. More specifically, compounds 7, 8, and 12 have superior membrane stability over indomethacin as a standard drug. At concentrations of 400, 600, 800, and 1000 mu g/mL, compound 12 demonstrated highly significant inhibition of RBC hemolysis with 95.2%, 97.3%, 98.9%, and 99.8%, respectively, compared with indomethacin at the same concentrations (94.5%, 97.3%, 98.5%, and 99.3%, respectively). COX assay indicated that compounds 7, 8, and 12 exhibited excellent COX-2 inhibition percentage (98.51 +/- 0.01, 98 +/- 0.01, and 98.97 +/- 0.06, respectively) at 150 mu g/mL using indomethacin (97.3 +/- 0.005). Furthermore, an investigation using molecular docking against COX-2 (pdb ID: 5IKT) was used to analyze the anti-inflammatory properties of all studied substances. When compared to indomethacin, molecular docking research showed a strong connection with the intended protein and an elevated docking score. According to in vitro anti-inflammatory action and computational approach, they show promise as a therapeutic candidate for the treatment of inflammatory diseases. A convenient green synthetic approach to the synthesis of a novel class of Spirotriazolotriazine derivatives with high Anti-inflammatory Activity.image