The current research was designed for selective green synthesis of N-(substituted)-2-(5-(1-(4-nitrophenylsulfonyl)piperidin-4-yl)-1,3,4-oxadiazol-2-ylthio)acetamide hybrids and their pharmacological applications to overcome the enzymatic diseases. Environment friendly sodium hydride as an activator was employed to catalyze the reaction for the synthesis of target compounds. Structural characterization was performed using spectroscopic techniques (¹H-NMR, ¹³C-NMR, IR, and EI-MS). All of the synthesized compounds were screened for acetylcholinesterase (AChE) and urease inhibition potential. Three compounds, 6a, 6k, and 6p, were found to be the best inhibitors of AChE enzyme and the whole array of compounds showed urease inhibition potential even better than the reference. Eserine was the reference standard for AChE inhibition and thiourea for urease inhibition. The computational and BSA binding studies were performed to explore the binding modes and interaction between synthesized compounds and respective enzymes. Molecular docking studies explained the presence of some hydrophobic interactions and have good correlation with calculated free energy of binding (DG binding). The finding of these new unique compounds having high potential to inhibit the activity of urease enzyme as compared to standard showing the prospective to replace the existing drugs and surprisingly effective addition of highly active drug discovery in the field of pharmacology against described enzyme.

N -Substituted-2-propanamide analogues of 1,3,4-oxadiazole have been synthesized using a multi-step synthetic protocol to explore new therapeutic anti-enzymatic agents. Herein, we have merged sulfonyl, piperidine, oxadiazole and amide into a single unit to synthesize a library of unique compounds, 8a–n . The molecular structures of all synthesized compounds were verified by ¹³ C-NMR, ¹ H-NMR, HRMS and IR spectroscopy. Furthermore, the compounds were screened for their inhibition potential against acetylcholinesterase (AChE), urease and lipoxygenase (LOX) enzymes. A considerable inhibition potential was observed for three compounds against LOX with quercetin as a reference standard, two compounds against urease with thiourea as a reference standard and two compounds against AChE with eserine as a reference standard. Through molecular docking investigations, we were able to correlate the overall impact and inhibition criteria by the structure–activity relationship via the interactions between synthesized compounds and active sites of enzymes. Pharmacodynamics, pharmacokinetics and in vivo studies may be investigated further for the most active compounds to substantiate them as potential anti-enzymatic medications.