Enzyme Inhibitory Kinetics and Molecular Docking Studies of Halo-Substituted Mixed Ester/Amide-Based Derivatives as Jack Bean Urease Inhibitors.

Muhammad Rashid,Zafar Iqbal,Sadia Roshan,Shazia Shamas,Qamar Abbas,Mubashir Hassan,Muhammad Hassham Hassan Bin Asad,Hummera Rafique,Zia Ur Rahman Qureshi,Zaman Ashraf

doi:10.1155/2020/8867407

Abstract

A series of halo-substituted mixed ester/amide-based analogues 4a-l have been prepared as jack bean urease inhibitor, which showed good to excellent inhibition of enzyme activity. The role of halo-substituted benzoyl moieties and alkyl substituted anilines in urease inhibitory kinetics was also investigated. The alkyl-substituted anilines 1a–b reacted with chloroacetyl chloride to afford intermediates 2a-b, which were then reacted with different halo-substituted benzoic acids 3a–f to prepare the title compounds 4a-l. The chemical structures of final products 4a-l were ascertained by FTIR, 1H NMR, 13C NMR, and mass spectra. The compound 4b showed remarkable activity with IC501.6 ± 0.2 nM, better than the standard thiourea having IC50472.1 ± 135.1 nM. The 2-chloro-substituted phenyl ring on one side of compound 4b and 4-isopropyl-substituted benzene on the other side play an essential role in inhibition of urease activity. Lineweaver–Burk plots (kinetics study) indicated about 4b derivative as a mixed type of inhibitor. The virtual screening performed against urease enzyme (PDBID 4H9M) showed that compounds 4b and 4e have binding energies of −7.8 and −7.9 Kcal/mol, respectively. Based upon our results, it was found that derivative 4b is a highly potent urease inhibitor, better than the standard thiourea.

Highlights

Urease (EC.3.5.1.5) is an enzyme of the amidohydrolase and phosphotriesterase family with nickel atoms present in their active binding sites
According to the FTIR spectrum, compounds 4a–l showed a characteristic peak of amide carbonyl (C=O) at 1640– 1685 cm−1, ester (C=O) at 1724–1745 cm−1, and secondary (N–H) stretching at 3250–3307 cm−1
The amide carbonyl absorption appeared at 1670 cm−1 in 2a and 1655 cm−1 in 2b and secondary (N–H) absorption at 3270 cm−1 in 2a and 3257 cm−1 in 2b in the FTIR spectrum, which confirmed the formation of 2a–b

Summary

Introduction

Urease (EC.3.5.1.5) is an enzyme of the amidohydrolase and phosphotriesterase family with nickel atoms present in their active binding sites. They catalyze hydrolysis of urea into carbamic acids which cleaved into carbon dioxide and ammonia (Figure 1) [1,2,3,4] which in turn increased the pH. The increased level of urease enzyme is associated with serious health problems like stomach cancer, peptic ulceration, and pyelonephritis [5,6,7]. The bacterial ureases increase the rate of urea hydrolysis which is associated with different biological disorders like. BioMed Research International O H2N NH2 Urea H HO OH Carbonic acid

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