Synthesis, biochemical characterization and molecular modeling studies of 5-(substituted benzylidene) pyrimidine-2,4,6-trione: Potential inhibitors of alkaline phosphatase

Abstract

Among the several forms of homodimeric protein enzymes, alkaline phosphatases are the major enzymes that play a critical part in a variety of biological activities and are responsible for the removal of phosphates from a variety of molecules. The alkaline phosphatase expression has been reported to be higher in numerous multifactorial disorders/syndromes and in cancer patients, making it an intriguing target for therapeutic research. Because of the substantial structural similarity between intestinal alkaline phosphatases (IAPs) and tissue-nonspecific alkaline phosphatases (TNAPs), only a few selective inhibitors have been discovered. Barbiturates are a privileged class of heterocyclic compounds that have the potential to act as efficient phosphatase inhibitors. In this context, a new series of 5-(substituted benzylidene) pyrimidine-2,4,6-triones (3a-j) was devised and easily synthesized using a simple and environmentally friendly Knovenagel condensation process. The FT-IR, 1H-NMR, and 13C-NMR were used to characterize the synthesized barbiturates. Furthermore, the compounds were examined for their ability to inhibit calf-intestinal alkaline phosphatases (CIAP). All the tested compounds showed inhibition of CIAP, however compound 5-(4-(Dimethylamino)benzylidene) pyrimidine-2,4,6(1H,3H,5H)-trion (3f) was shown to be the most effective and best CIAP inhibitor, with an IC50 value of 0.29±0.02 µM. To test their stability and binding affinities within the active pocket of the CIAP, the structure activity relationship, molecular docking studies, and density function theory (DFTs) calculations were performed. The Global chemical reactivity descriptors were assessed through DFTs. Docking study indicated that the most active inhibitor (3f) exhibited significant interactions within the binding pockets of intestinal alkaline phosphatase. As a result, the screened barbiturates derivatives provided an excellent platform for the subsequent development of alkaline phosphatase inhibitors.A new series of 5-(substituted benzylidene) pyrimidine-2,4,6-triones (3a-j) was devised and easily synthesized using a simple and environmentally friendly Knovenagel condensation reaction. The FT-IR, 1H-NMR/13C-NMR were used to characterize the synthesized compounds. The compounds were then examined for their ability to inhibit the calf-intestinal alkaline phosphatases (CIAP). All the tested compounds showed inhibition of CIAP, however compound 5-(4-(Dimethylamino)benzylidene) pyrimidine-2,4,6(1H,3H,5H)-trion 3f was shown to be the most effective and best CIAP inhibitor, with an IC50 value of 0.29±0.02 µM. To test their stability and binding affinities within the active pocket of the CIAP target site, the structure activity relationship, molecular docking studies, and density function theory calculations were performed. Based on the results of the docking investigation, the most effective inhibitor 3f was found to have strong interactions within the binding pockets of intestinal alkaline phosphatase. Global chemical reactivity descriptors were assessed by using density functional calculation. From In vitro as well as computational studies, the derivatives of barbiturates that were screened proved to be a suitable basis for the subsequent research and development of alkaline phosphatase inhibitors.