Streptococcus pneumononiae gyrase ATPase: Development and validation of an assay for inhibitor discovery and characterization

Miller, J. Richard; Herberg, John T.; Tomilo, Mark; McCroskey, Mark C.; Feilmeier, Bradley

doi:10.1016/j.ab.2007.02.029

Cited by 8 publications

(6 citation statements)

References 33 publications

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“…Microcalorimetry provides an accurate thermodynamic evaluation of the interaction in terms of enthalpy change, entropy change, and Gibbs Free energy change along with the stoichiometry and binding affinity from a titration. ITC is an effective tool to thermodynamically characterize the binding of small molecules to macromolecules. −, Figure A depicts the representative ITC thermogram for the complexation of the 12b with EcTopo 1A. In the upper panel, the raw data for the sequential injection of EcTopo 1A into 12b are depicted.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Biological Evaluation of Novel Bisbenzimidazoles as Escherichia coli Topoisomerase IA Inhibitors and Potential Antibacterial Agents

et al. 2014

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Novel bisbenzimidazole inhibitors of bacterial type IA topoisomerase are of interest for the development of new antibacterial agents that are impacted by target-mediated cross resistance with fluoroquinolones. The present study demonstrates the successful synthesis and evaluation of bisbenzimidazole analogues as Escherichia coli topoisomerase IA inhibitors. 5-(4-Propylpiperazin-1-yl)-2-[2'-(4-ethoxyphenyl)-5'-benzimidazolyl]benzimidazole (12b) showed significant relaxation inhibition activity against EcTopo 1A (IC50 = 2 ± 0.005 μM) and a tendency to chelate metal ion. Interestingly, these compounds did not show significant inhibition of E. coli DNA gyrase and hTop 1 even up to 100 μM. Compound 12b has shown lowest MIC against E. coli strains among 24 compounds evaluated. The binding affinity constant and binding free energy of 12b with EcTopo 1A was observed 6.8 × 10(6) M(-1) and -10.84 kcal mol(-1) from isothermal titration calorimetry (ITC), respectively. In vivo mouse systemic infection and neutropenic thigh model experimental results confirmed the therapeutic efficacy of 12b, suggesting further development of this class of compounds as antibacterial agents.

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Section: Resultsmentioning

confidence: 99%

Synthesis and Biological Evaluation of Novel Bisbenzimidazoles as Escherichia coli Topoisomerase IA Inhibitors and Potential Antibacterial Agents

et al. 2014

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“…148−151 This mineral phosphate release can also be assessed by use of 7-methyl-6-thioguanosine and purine nucleoside phosphorylase, which leads to the occurrence of a product with increased absorbance at 360 nm. 132,152,153 The binding of tritium-labeled [ 3 H]dihydronovobiocin to a biotin-labeled 43-kDa fragment of GyrB (biotin−GyrB43) was used as an assay amenable to high-throughput screening for DNA gyrase inhibitors. 154 Fluorescence polarization-based assays are also possible since derivatization, with a fluorescent component, of known inhibitors can provide probes that retain an affinity for the type IIA topoisomerases.…”

Section: Assays For Bacterial Type Iia Topoisomerase Inhibitionmentioning

confidence: 99%

Non-quinolone Inhibitors of Bacterial Type IIA Topoisomerases: A Feat of Bioisosterism

2013

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“…Gyrase is the cellular target of several different classes of bacteriocidal agents, including the highly-successful fluoroquinolone antibiotics, coumarins and cyclothialidines that are currently undergoing drug development, as well as a range of naturally-occurring bacterial toxins [22, 97]. Much of the current research on gyrase inhibitors focuses on modifying existing scaffolds to address issues with drug resistance, toxic side effects and poor cellular penetration, as well as to broaden the range of bacterial species that compounds are active against [22, 97, 98]. Structure-aided design and virtual screening techniques have proven somewhat successful in delivering new classes of gyrase inhibitors, some of which show broad-spectrum antibacterial activity and reduced rates of resistance relative to exisiting drugs [99, 100].…”

Section: Existing Inhibitors and Current Screening Strategiesmentioning

confidence: 99%

Architecture and Conservation of the Bacterial DNA Replication Machinery, an Underexploited Drug Target

Robinson¹,

Causer²,

Dixon

2012

CDT

105

124

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New antibiotics with novel modes of action are required to combat the growing threat posed by multi-drug resistant bacteria. Over the last decade, genome sequencing and other high-throughput techniques have provided tremendous insight into the molecular processes underlying cellular functions in a wide range of bacterial species. We can now use these data to assess the degree of conservation of certain aspects of bacterial physiology, to help choose the best cellular targets for development of new broad-spectrum antibacterials.DNA replication is a conserved and essential process, and the large number of proteins that interact to replicate DNA in bacteria are distinct from those in eukaryotes and archaea; yet none of the antibiotics in current clinical use acts directly on the replication machinery. Bacterial DNA synthesis thus appears to be an underexploited drug target. However, before this system can be targeted for drug design, it is important to understand which parts are conserved and which are not, as this will have implications for the spectrum of activity of any new inhibitors against bacterial species, as well as the potential for development of drug resistance. In this review we assess similarities and differences in replication components and mechanisms across the bacteria, highlight current progress towards the discovery of novel replication inhibitors, and suggest those aspects of the replication machinery that have the greatest potential as drug targets.

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Streptococcus pneumononiae gyrase ATPase: Development and validation of an assay for inhibitor discovery and characterization

Cited by 8 publications

References 33 publications

Synthesis and Biological Evaluation of Novel Bisbenzimidazoles as Escherichia coli Topoisomerase IA Inhibitors and Potential Antibacterial Agents

Synthesis and Biological Evaluation of Novel Bisbenzimidazoles as Escherichia coli Topoisomerase IA Inhibitors and Potential Antibacterial Agents

Non-quinolone Inhibitors of Bacterial Type IIA Topoisomerases: A Feat of Bioisosterism

Architecture and Conservation of the Bacterial DNA Replication Machinery, an Underexploited Drug Target

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