Understanding the enzymatic inhibition of intestinal alkaline phosphatase by aminophenazone-derived aryl thioureas with aided computational molecular dynamics simulations: synthesis, characterization, SAR and kinetic profiling

The work presented in this paper aims toward the synthesis of aryl thiourea derivatives4a-lof pyrazole based nonsteroidal anti-inflammatory drug named 4-aminophenazone, as potential inhibitors of intestinal alkaline phosphatase enzyme. The screening of synthesized target compounds4a-lfor unraveling the anti-inflammatory potential against calf intestinal alkaline phosphatase gives rise to lead member4cpossessing IC(50)value 0.420 +/- 0.012 mu M, many folds better than reference standard used (KH2PO4IC50 = 2.8 +/- 0.06 mu M andl-phenylalanine IC50 = 100 +/- 3.1 mu M). SAR for unfolding the active site binding pocket interaction along with the mode of enzyme inhibition based on kinetic studies is carried out which showed non-competitive binding mode. The enzyme inhibition studies were further supplemented by molecular dynamic simulations for predicting the protein behavior against active inhibitors4cand4gduring docking analysis. The preliminary toxicity of the synthesized compounds was determined by using brine shrimp assay. This work also includes detailed biochemical analysis along with RO5 parameters for all the newly synthesized drug derivatives4a-l. [GRAPHICS] .

Automated summary

The work focuses on synthesizing aryl thiourea derivatives of pyrazole-based nonsteroidal anti-inflammatory drug as potential inhibitors of intestinal alkaline phosphatase enzyme. The lead compound 4c showed significant inhibitory activity with a non-competitive binding mode, as revealed by kinetic studies and molecular dynamic simulations during docking analysis. Preliminary toxicity and biochemical analysis were also conducted for all newly synthesized drug derivatives4a-l.