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

• Published in: Molecular diversity 2021-08, Vol.25 (3), p.1701-1715
• Main Authors: Khurshid, Asma, Saeed, Aamer, Ashraf, Zaman, Abbas, Qamar, Hassan, Mubashir
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - chemistry; Aminopyrine - analogs & derivatives; Aminopyrine - chemistry; Binding Sites; Biochemistry; Biomedical and Life Sciences; Chemical Phenomena; Chemistry Techniques, Synthetic; Enzyme Inhibitors - chemical synthesis; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Enzymes; Humans; Kinetics; Life Sciences; Molecular Docking Simulation; Molecular Dynamics Simulation; Molecular Structure; Organic Chemistry; Original Article; Pharmacy; Phosphatase; Polymer Sciences; Protein Binding; Solvents; Spectrum Analysis; Structure-Activity Relationship; Thiourea - chemical synthesis; Thiourea - chemistry; Thiourea - pharmacology
• ISSN: 1381-1991; 1573-501X
• DOI: 10.1007/s11030-020-10136-9

The work presented in this paper aims toward the synthesis of aryl thiourea derivatives 4a – l of pyrazole based nonsteroidal anti-inflammatory drug named 4-aminophenazone, as potential inhibitors of intestinal alkaline phosphatase enzyme. The screening of synthesized target compounds 4a – l for unraveling the anti-inflammatory potential against calf intestinal alkaline phosphatase gives rise to lead member 4c possessing IC 50 value 0.420 ± 0.012 µM, many folds better than reference standard used (KH 2 PO 4 IC 50   =  2.8 ± 0.06 µM and l -phenylalanine IC 50  = 100 ± 3.1 µM). 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 inhibitors 4c and 4g during 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 derivatives 4a – l . Graphic abstract