BIOSCIENCE REPORTS
·
2018
·Vol. 38
·期 2
·
IF 2.899
Wang Qian,He Yuan,Lu Rui,Wang Wen-Ming,Yang Ke-Wu,Fan Hai Ming,Jin Yi,Blackburn G. Michael
The New Delhi metallo-β-lactamase (NDM-1) is an important clinical target for antimicrobial research, but there are insufficient clinically useful inhibitors and the details of NDM-1 enzyme catalysis remain unclear. The aim of this work is to provide a thermodynamic profile of NDM-1 catalysed hydrolysis of β-lactams using an isothermal titration calorimetry (ITC) approach and to apply this new method to the identification of new low-molecular-weight dicarboxylic acid inhibitors. The results reveal that hydrolysis of penicillin G and imipenem by NDM-1 share the same thermodynamic features with a significant intrinsic enthalpy change and the release of one proton into solution, while NDM-1 hydrolysis of cefazolin exhibits a different mechanism with a smaller enthalpy change and the release of two protons. The inhibitory constants of four carboxylic acids are found to be in the micromolar range. The compounds pyridine-2,6-dicarboxylic acid and thiazolidine-2,4-dicarboxylic acid show the best inhibitory potency and are confirmed to inhibit NDM-1 using a clinical strain of Escherichia coli. The pyridine compound is further shown to restore the susceptibility of this E. coli strain to imipenem, at an inhibitor concentration of 400 μM, while the thiazoline compound also shows a synergistic effect with imipenem. These results provide valuable information to enrich current understanding on the catalytic mechanism of NDM-1 and to aid the future optimisation of β-lactamase inhibitors based on these scaffolds to tackle the problem of antibiotic resistance.
研究领域:
微生物学
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药理学
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生物化学
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DOI
PubMed
高影响因子
JOURNAL OF ANTIBIOTICS
·
2021
·Vol. 74
·期 9
·574-579
·
IF 2.649
Ge Ying,Kang Peng-Wei,Li Jia-Qi,Gao Han,Zhai Le,Sun Le-Yun,Chen Cheng,Yang Ke-Wu
The superbug infection caused by metallo-β-lactamases (MβLs) carrying drug-resistant bacteria, specifically, New Delhi metallo-β-lactamase (NDM-1) has become an emerging threat. In an effort to develop novel inhibitors of NDM-1, thirteen thiosemicarbazones ( 1a - 1m ) were synthesized and assayed. The obtained molecules specifically inhibited NDM-1, with an IC 50 in the range of 0.88–20.2 µM, and 1a and 1f were found to be the potent inhibitors (IC 50 = 1.79 and 0.88 μM) using cefazolin as substrate. ITC and kinetic assays indicated that 1a irreversibly and non-competitively inhibited NDM-1 in vitro. Importantly, MIC assays revealed that these molecules by themselves can sterilize NDM-producing clinical isolates EC01 and EC08, exhibited 78-312-fold stronger activities than the cefazolin. MIC assays suggest that 1a (16 μg ml −1 ) has synergistic antimicrobial effect with ampicillin, cefazolin and meropenem on E. coli producing NDM-1, resulting in MICs of 4-32-, 4-32-, and 4-8-fold decrease, respectively. These studies indicate that the thiosemicarbazide is a valuable scaffold for the development of inhibitors of NDM-1 and NDM-1 carrying drug-resistant bacteria.
研究领域:
微生物学
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药理学
·
感染病学
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DOI
PubMed
高影响因子
Experimental and Therapeutic Medicine
·
2019
·Vol. 18
·期 4
·2924-2932
·
IF 1.448
Jingjing Wang,Yongzhong Ning,Shu Li,Yun Wang,Jinhua Liang,Chunming Jin,Hairun Yan,Yongcun Huang
Acinetobacter baumannii is an important cause of hospital‑acquired, multidrug‑resistant (MDR) infections occurring worldwide. Anti‑microbial combination regimens may be the only feasible treatment option for affected patients. In the present study, the efficacy of the combined therapy of meropenem with colistin, ampicillin‑sulbactam, tazobactam and vancomycin against clinical strains of MDR A. baumannii was determined. Anti‑microbial susceptibility testing was performed and resistance genes were characterized by a multiplex polymerase chain reaction (PCR)‑reverse line blot assay. The genetic background of New Delhi metallo‑β‑lactamase 1 (NDM‑1) was analysed by primer walking. The presence of NDM‑1 was detected using the modified Hodge test and the EDTA‑combined disk test. To screen for synergistic drug effects, the fractional inhibitory concentration index was calculated using a checkerboard assay. The results of the PCR as well as the sequence analyses suggested that NDM‑1 was located downstream of the ISAba125 element. In addition, a synergistic effect was determined for meropenem + vancomycin, meropenem + tazobactam and meropenem + ampicillin + sulbactam in two strains each, and in four strains for meropenem + colistin. A total of five A. baumannii strains with resistance to numerous antibiotics and carrying numerous resistance genes were identified. In the strains of A. baumannii, the NDM‑1 gene was integrated in a transposon structure with a copy of the ISAba125 insertion sequence. However, the genetic background was not identical among the different species and strains. The genetic variability of NDM‑1 may facilitate the rapid dissemination of this gene. In conclusion, meropenem may enhance the efficacy of antibiotics in A. baumannii strains with NDM‑1‑associated MDR.
研究领域:
微生物学
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传染病学
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临床医学
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DOI
PubMed
高影响因子
Biomed Research International
·
2022
·
IF 3.246
Aparna Vasudevan,Dinesh Kumar Kesavan,Liang Wu,Zhaoliang Su,Shengjun Wang,Mohan Kumar Ramasamy,Waheeta Hopper,Huaxi Xu
Antibiotic resistance is one of the significant problems globally; there is an increase in resistance with introducing every new class of antibiotics. Further, this has become one of the reasons for arising of new resistance mechanisms in Acinetobacter baumannii. In this study, we have screened natural compounds as a possible inhibitor against the NDM-1 β-lactamase enzyme from A. baumannii using a combination of in silico methods and in vitro evaluation. The database of natural compounds was screened against NDM-1 protein, using Glide docking, followed by QM-polarised ligand docking (QPLD). When the screened hits were validated in vitro, withaferin A and mangiferin had good IC50 values in reducing the activity of NDM-1 enzymes, and their fractional inhibitory concentration index (FICI) was ascertained in combination with imipenem. The withaferin A and mangiferin-NDM-1 docking complexes were analyzed for structural stability by molecular dynamic simulation analysis using GROMACS for 100 ns. The molecular properties of the natural compounds were then calculated using density functional theory (DFT). Withaferin A and mangiferin showed promising inhibitory activity and can be a natural compound candidate inhibitor synergistically used along with carbapenems against NDM-1 producing A. baumannii.
研究领域:
微生物学
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药物化学
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计算生物学
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DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2018
·
IF 3.929
Jianpeng Su,Jiayun Liu,Cheng Chen,Yuejuan Zhang,Kewu Yang
The superbug infection caused by New Delhi metallo-β-lactamase (NDM-1) has grown into an emerging threat, labelling and inhibition of NDM-1 has proven challenging due to its shuttling between pathogenic bacteria. Here, we report a potent covalent scaffold, ebsulfur, for targeting the protein in vitro and in vivo . Enzymatic kinetic study indicated that eighteen ebsulfurs gained except 1a – b and 1f inhibited NDM-1, exhibiting an IC 50 value ranging of 0.16–9 μM, and 1g was found to be the best, dose- and time-dependent inhibitor with an IC 50 of 0.16 μM. Also, these ebsulfurs effectively restored the antibacterial activity of cefazolin against E. coli expressing NDM-1, and the best effect was observed to be from 1g , 1i and 1n , resulting in an 256-fold reduction in MIC of the antibiotic at a dose of 16 μg/mL. The equilibrium dialysis study implied that the ebsulfur disrupted the coordination of one Zn(II) ion at active site of NDM-1. Labelling of NDM-1 using a constructed fluorescent ebsulfur Ebs - R suggested that the inhibitor covalently bound to the target through SDS-PAGE analysis in vitro . Also, labelling NDM-1 in living E. coli cells with Ebs - R by confocal microscopic imaging showed the real-time distribution change process of intracellular recombinant protein NDM-1. Moreover, the cytotoxicity of these ebsulfurs against L929 mouse fibroblastic cells was tested, and their capability to restore antibacterial activity of antibiotic against clinical strains E. coli EC08 producing NDM-1 was determined. The ebsulfur scaffold proposed here is valuable for development of the covalent irreversible inhibitors of NDM-1, and also for labelling the target in vitro and in vivo .
研究领域:
微生物学
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药理学
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生物化学
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DOI
PubMed
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2020
·Vol. 167
·期 6
·067525
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IF 3.721
Yi Xiu,Ruiping Luo,Baoqing Han,Lu Liu,Hongsu Wang
There is an urgent need to develop convenient, rapid and sensitive detection methods to determine antibiotic residues in food. New Delhi metal-β-lactamase-1 (NDM-1) is a β-lactamase enzyme that catalyzes removal of all β-lactam antibiotics other than aztreonam via a spectral hydrolysis mechanism. In the present study, a new biosensor based on NDM-1 was designed and tested to detect penicillin antibiotics namely penicillin sodium (Pen G) and ampicillin sodium (AMP) without using any pH redox probe. The glassy carbon electrode (GCE) was modified by electro-polymerization of methylene blue (PMB), and the biosensor (NDM-1/PMB/GCE) was prepared by immobilizing NDM-1 enzyme with glutaraldehyde (GA). Penicillin was detected by differential pulse voltammetry (DPV). This study seems to be one of the first kind to apply NDM-1 enzyme to detect antibiotic residues in food. It can be used repeatedly and can successfully detect penicillin in actual milk samples by spike recovery experiment.
研究领域:
食品科学
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生物化学
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DOI
高影响因子
Chemical Biology & Drug Design
·
2021
·Vol. 99
·期 2
·362-372
·
IF 2.817
Jia-Zhu Chigan,Jia-Qi Li,Huan-Huan Ding,Yin-Sui Xu,Lu Liu,Cheng Chen,Ke-Wu Yang
Bacterial resistance caused by metallo-β-lactamases (MβLs) has become an emerging public health threat, and the development of MβLs inhibitor is an effective way to overcome the resistance. In this study, thirteen novel O -aryloxycarbonyl hydroxamates were constructed and assayed against MβLs. The obtained molecules specifically inhibited imipenemase-1 (IMP-1) and New Delhi metallo-β-lactamase-1, exhibiting an IC 50 value in the range of 0.10–18.42 and 0.23–22.33 μM, respectively. The hydroxamate 5 was found to be the most potent inhibitor, with an IC 50 of 0.1 and 0.23 μM using meropenem and cefazolin as substrates. ICP-MS analysis showed that 5 did not coordinate to the Zn(II) ions at the active site of IMP-1, while the rapid dilution, thermal shift and MALDI-TOF assays revealed that the hydroxamate formed a covalent bond with the enzyme. Cytotoxicity assays indicated that the hydroxamates have low toxicity in MCF-7 cells. This work provided a potent scaffold for the development of MβLs inhibitors.
研究领域:
微生物学
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药物化学
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感染病学
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2023
·
IF 4.8
Hongyu Gan,Yaowen Liang,Yun Tang,Manna Huang,Yiqian Wan
A practical electrochemical method for the determination of the inhibitory activity of New Delhi metallo-β-lactamase-1 (NDM-1) inhibitors was established with a bare pencil graphite electrode (PGE) as the working electrode and imipenem as the substrate. To validate the method, the well-known NDM-1 inhibitors EDTA and L -captopril were selected for the corresponding assays with IC 50 values of 1.22 ± 0.20 μM and 22.92 ± 1.19 μM, respectively. These results were consistent with those (5.48 ± 0.32 μM for EDTA and 69.13 ± 6.01 μM for L- captopril) from the UV–vis high-throughput assay method. Further investigation regarding the steady kinetics of a typical competitive inhibitor of L -captopril showed that the two methods provided K i values at the same level, i.e., they are 41.5 ± 3.8 μM (DPV-PGE) and 52.5 ± 2.5 μM (UV–vis), respectively.
研究领域:
生物化学
·
药物化学
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产品关联
DOI
高影响因子
CHEMICAL & PHARMACEUTICAL BULLETIN
·
2021
·Vol. 69
·期 12
·1179-1183
·
IF 1.645
Taichi Kamo,Keiichi Kuroda,Shota Kondo,Usaki Hayashi,Satoshi Fudo,Tomoki Yoneda,Akiko Takaya,Michiyoshi Nukaga,Tyuji Hoshino
Metallo-β-lactamases (MBLs) are significant threats to humans because they deteriorate many kinds of β-lactam antibiotics and are key enzymes responsible for multi-drug resistance of bacterial pathogens. As a result of in vitro screening, two compounds were identified as potent inhibitors of two kinds of MBLs: imipenemase (IMP-1) and New Delhi metallo-β-lactamase (NDM-1). The binding structure of one of the identified compounds was clarified by an X-ray crystal analysis in complex with IMP-1, in which two possible binding poses were observed. Molecular dynamics (MD) simulations were performed by building two calculation models from the respective binding poses. The compound was stably bound to the catalytic site during the simulation in one pose. The binding model between NDM-1 and the compound was constructed for MD simulation. Calculation results for NDM-1 were similar to those of IMP-1. The simulation suggested that the binding of the identified inhibitory compound was also durable in the catalytic site of NDM-1. The compound will be a sound basis for the development of the inhibitors for MBLs.
研究领域:
微生物学
·
结构生物学
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药物化学
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DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2020
·
IF 4.831
De-Yun Cui,Yi Yang,Meng-Meng Bai,Jiang-Xue Han,Cong-Cong Wang,Hong-Tao Kong,Bo-Yuan Shen,Da-Chao Yan,Chun-Ling Xiao,Yi-Shuang Liu,En Zhang
New Delhi Metallo-β-lactamase-1 (NDM-1), a Zn (II)-dependent enzyme, can catalyze the hydrolysis of almost all β-lactam antibiotics including carbapenems, resulting in bacterial antibiotic resistance, which threatens public health globally. Based on our finding that H 2 dedpa is as an efficient NDM-1 inhibitor, a series of H 2 dedpa derivatives was systematically prepared. These compounds exhibited significant activity against NDM-1, with IC 50 values 0.06–0.94 μM. In vitro , compounds 6k and 6n could restore the activity of meropenem against Klebsiella pneumoniae , Escherichia coli and Proteus mirabilis possessing either NDM or IMP. In particular, the activity of meropenem against E. coli producing NDM-4 could be improved up to 5333 times when these two compounds were used. Time–kill cell-based assays showed that 99.9% of P. mirabilis were killed when treated with meropenem in combination with compound 6k or 6n . Furthermore, compounds 6k and 6n were nonhemolytic (HC 50 > 1280 μg/mL) and showed low toxicity toward mammalian (HeLa) cells. Mechanistic studies indicated that compounds 6k and 6n inhibit NDM-1 by chelating the Zn 2+ ion of the enzyme.
研究领域:
微生物学
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药物化学
·
感染病学
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DOI
高影响因子
Materials Today Bio
·
2026
·
IF 11
Zhaoyou Chu,Shu Fang,Wanyue Fu,Jun Liang,Yayun Wu,Ling Xu,Hanqing Zhang,Jiyang Wang,Haisheng Qian,Min Shao
New Delhi metallo-β-lactamase (NDM)-producing drug-resistant bacteria often form stubborn biofilms and cause severe pneumonia, resulting in conventional antibiotic failure. In this study, we developed gelatin-genipin microspheres co-loaded with EDTA and meropenem (GEM) for targeted treatment of NDM pneumonia. EDTA exerted dual functions: it chelated Zn 2+ to inactivate NDM enzyme and restore meropenem activity (FIC = 0.046875), while also disrupting bacterial biofilms through chelation. GEM microspheres enable accelerated drug release in the artificial phlegm environment. In a mouse model of NDM pneumonia, GEM microspheres can enhance the recruitment of neutrophils and M1 cells at the early stage (12 h) through bacterial clearance, modulate the secretion of interferon-γ by CD8 + T cells and natural killer cells, and reshape the pulmonary immune microenvironment. Furthermore, GEM treatment reduced pathological pulmonary immune cell infiltration and excessive inflammation at late time points (days 3-7). Taken together, GEM microspheres integrate biofilm disruption, enzyme inhibition, synergistic bactericidal effects, and immunoregulation, representing a promising strategy against NDM drug-resistant bacterial pneumonia.
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PubMed
高影响因子
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