Synthesis of 1,2,4,5-Tetrasubstituted Imidazoles via ultrasound-assisted method and in vitro assessment of their anti-inflammatory and Hemocompatibility activity

The 1,2,4,5-tetrasubstituted imidazole core is increasingly recognized for its broad pharmacological effects, particularly its anti-inflammatory properties. This work employed a one-pot, four-component condensation reaction using benzil, substituted aromatic aldehydes, aromatic amines, and ammonium acetate, catalysed by sulfuric acid, to synthesise a series of tetrasubstituted imidazole derivatives (M1–M10). Ultrasound-assisted synthesis was more efficient than traditional reflux methods, providing shorter reaction times and higher yields. FT-IR, 1 H NMR, 13 C NMR, and mass spectrometry analyses confirmed the structures of the products. Selected derivatives (M1, M3, M7, and M10) were evaluated for their anti-inflammatory and hemocompatibility properties using proteinase inhibition, albumin denaturation, and hemolysis assays. Diclofenac sodium served as the reference drug for anti-inflammatory tests. Among the synthesized compounds, M10 exhibited the highest activity, with 69.00% protein inhibition and 82.39% inhibition of albumin denaturation at 1000 μg/mL, compared with diclofenac sodium (61.69% and 83.33%, respectively). Compounds M1 and M7 also displayed notable effects, with protein inhibition values of 62.16% and 63.16%, and albumin denaturation inhibition values of 77.12% and 67.67%, respectively. All tested compounds demonstrated excellent hemocompatibility, with hemolysis rates ranging from 0.74% to 2.06%, well below the 5% safety threshold. These results indicate that the synthesized 1,2,4,5-tetrasubstituted imidazole derivatives exhibit significant in vitro anti-inflammatory activity and favorable hemocompatibility, demonstrating the practical effectiveness of the ultrasound-assisted synthesis method. • Ultrasound-assisted one-pot synthesis of 1,2,4,5-tetrasubstituted imidazoles ( M1–M10 ). • Greener protocol with shorter reaction time and higher yield than reflux. • Structures confirmed by FT-IR, 1 H NMR, 13 C NMR, and mass analyses. • M1, M3, M7, M10 showed strong anti-inflammatory activity vs diclofenac. • All tested compounds showed <5% hemolysis, indicating good hemocompatibility.