Novel 3-Amino-2-methylquinazolinone NF-κB Inhibitors: Synthesis and Potential Anti-Inflammatory Function

The canonical NF-κB pathway is a major regulator of inflammatory responses, and its persistent activation has been linked to chronic inflammatory diseases and cancer development. It is therefore important to develop effective NF-κB inhibitors that can potentially be utilized in further preclinical studies. In this study, we have synthesized various quinazolinone compounds using novel methodologies. We specifically utilized a two-step protocol comparing an established protocol to a modified alternative procedure and then successfully established a novel one-pot multi-component reaction. All quinazolinones were tested based on their capacity to inhibit tumor necrosis factor-induced NF-κB activation using a commercial transgenic HeLa reporter cell-line. Among the eighteen molecules tested, two compounds affected the NF-κB-dependent luciferase activity, the 5o (3-amino-6-hydroxy-2-methylquinazolin-4(3H)-one) and 10 (N-(6-bromo-2-methyl-4-oxoquinazolin-3(4H)-yl)-4-nitrobenzamide). Only the 5o though, bearing OH at R¹ (6 position), attenuated the production of pro-inflammatory interleukin 6 in lipopolysaccharide-stimulated human primary macrophage cultures. Molecular docking analysis provides supportive computational evidence that 5o could interact with the p65/DNA complex and therefore could potentially be developed for further studies that aim to inhibit the canonical NF-κB pathway.