Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor
Inhibition of β-lactamases presents a promising strategy to restore the β-lactams antibacterial activity to resistant bacteria. In this work, we found that aromatic carboxyl substituted 2-triazolylthioacetamides <b>1a</b>−<b>j</b> inhibited VIM-2,...
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doaj-89087979630147f5b60c6b5fa601368e2020-11-25T00:33:36ZengMDPI AGBiomolecules2218-273X2020-01-011017210.3390/biom10010072biom10010072Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) InhibitorYang Xiang0Yue-Juan Zhang1Ying Ge2Yajun Zhou3Cheng Chen4Weixiao Yuan Wahlgren5Xiangshi Tan6Xi Chen7Ke-Wu Yang8Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaKey Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaKey Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaDepartment of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, ChinaKey Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaDepartment of Chemistry and Molecular Biology, University of Gothenburg, Box 462, S-40530 Gothenburg, SwedenDepartment of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, ChinaKey Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaKey Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University, Xi’an 710127, ChinaInhibition of β-lactamases presents a promising strategy to restore the β-lactams antibacterial activity to resistant bacteria. In this work, we found that aromatic carboxyl substituted 2-triazolylthioacetamides <b>1a</b>−<b>j</b> inhibited VIM-2, exhibiting an IC<sub>50</sub> value in the range of 20.6−58.6 μM. The structure-activity relationship study revealed that replacing the aliphatic carboxylic acid with aromatic carboxyl improved the inhibitory activity of 2-triazolylthioacetamides against VIM-2. <b>1a</b>−<b>j</b> (16 mg/mL) restored the antibacterial activity of cefazolin against <i>E. coli</i> cell expressing VIM-2, resulting in a 4−8-fold reduction in MICs. The isothermal titration calorimetry (ITC) characterization suggested that the primary binding 2-triazolylthioacetamide (<b>1b</b>, <b>1c</b>, or <b>1h</b>) to VIM-2 was a combination of entropy and enthalpy contributions. Further, the crystal structure of VIM-2 in complex with <b>1b</b> was obtained by co-crystallization with a hanging-drop vapour-diffusion method. The crystal structure analysis revealed that <b>1b</b> bound to two Zn(II) ions of the enzyme active sites, formed H-bound with Asn233 and structure water molecule, and interacted with the hydrophobic pocket of enzyme activity center utilizing hydrophobic moieties; especially for the phenyl of aromatic carboxyl which formed π-π stacking with active residue His263. These studies confirmed that aromatic carboxyl substituted 2-triazolylthioacetamides are the potent VIM-2 inhibitors scaffold and provided help to further optimize 2-triazolylthioacetamides as VIM-2 even or broad-spectrum MβLs inhibitors.https://www.mdpi.com/2218-273X/10/1/72antibiotic resistancemetallo-β-lactamase vim-2 inhibitor2-triazolylthioacetamidesthermodynamicscrystallographic study |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yang Xiang Yue-Juan Zhang Ying Ge Yajun Zhou Cheng Chen Weixiao Yuan Wahlgren Xiangshi Tan Xi Chen Ke-Wu Yang |
spellingShingle |
Yang Xiang Yue-Juan Zhang Ying Ge Yajun Zhou Cheng Chen Weixiao Yuan Wahlgren Xiangshi Tan Xi Chen Ke-Wu Yang Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor Biomolecules antibiotic resistance metallo-β-lactamase vim-2 inhibitor 2-triazolylthioacetamides thermodynamics crystallographic study |
author_facet |
Yang Xiang Yue-Juan Zhang Ying Ge Yajun Zhou Cheng Chen Weixiao Yuan Wahlgren Xiangshi Tan Xi Chen Ke-Wu Yang |
author_sort |
Yang Xiang |
title |
Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor |
title_short |
Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor |
title_full |
Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor |
title_fullStr |
Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor |
title_full_unstemmed |
Kinetic, Thermodynamic, and Crystallographic Studies of 2-Triazolylthioacetamides as Verona Integron-Encoded Metallo-β-Lactamase 2 (VIM-2) Inhibitor |
title_sort |
kinetic, thermodynamic, and crystallographic studies of 2-triazolylthioacetamides as verona integron-encoded metallo-β-lactamase 2 (vim-2) inhibitor |
publisher |
MDPI AG |
series |
Biomolecules |
issn |
2218-273X |
publishDate |
2020-01-01 |
description |
Inhibition of β-lactamases presents a promising strategy to restore the β-lactams antibacterial activity to resistant bacteria. In this work, we found that aromatic carboxyl substituted 2-triazolylthioacetamides <b>1a</b>−<b>j</b> inhibited VIM-2, exhibiting an IC<sub>50</sub> value in the range of 20.6−58.6 μM. The structure-activity relationship study revealed that replacing the aliphatic carboxylic acid with aromatic carboxyl improved the inhibitory activity of 2-triazolylthioacetamides against VIM-2. <b>1a</b>−<b>j</b> (16 mg/mL) restored the antibacterial activity of cefazolin against <i>E. coli</i> cell expressing VIM-2, resulting in a 4−8-fold reduction in MICs. The isothermal titration calorimetry (ITC) characterization suggested that the primary binding 2-triazolylthioacetamide (<b>1b</b>, <b>1c</b>, or <b>1h</b>) to VIM-2 was a combination of entropy and enthalpy contributions. Further, the crystal structure of VIM-2 in complex with <b>1b</b> was obtained by co-crystallization with a hanging-drop vapour-diffusion method. The crystal structure analysis revealed that <b>1b</b> bound to two Zn(II) ions of the enzyme active sites, formed H-bound with Asn233 and structure water molecule, and interacted with the hydrophobic pocket of enzyme activity center utilizing hydrophobic moieties; especially for the phenyl of aromatic carboxyl which formed π-π stacking with active residue His263. These studies confirmed that aromatic carboxyl substituted 2-triazolylthioacetamides are the potent VIM-2 inhibitors scaffold and provided help to further optimize 2-triazolylthioacetamides as VIM-2 even or broad-spectrum MβLs inhibitors. |
topic |
antibiotic resistance metallo-β-lactamase vim-2 inhibitor 2-triazolylthioacetamides thermodynamics crystallographic study |
url |
https://www.mdpi.com/2218-273X/10/1/72 |
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