Evaluation of 2,6-difluoro-3-(oxazol-2-ylmethoxy)benzamide chemotypes as Gram-negative FtsZ inhibitors

Rosado-Lugo, Jesus D.; Sun, Yangsheng; Banerjee, Anamika; Cao, Yanlu; Datta, Pran K.; Zhang, Yongzheng; Yuan, Zhen; Parhi, Ajit K.

doi:10.1038/s41429-022-00531-9

Cited by 12 publications

(14 citation statements)

References 27 publications

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“…More recently, two molecules, TXA6101 and TXY6129, with substituted 2,6difluorobenzamide scaffold, have been shown to inhibit the polymerization of both E. coli and Klebsiella pneumoniae FtsZ. Moreover, despite being substrates for efflux pumps, TXA6101 induced morphological changes in K. pneumoniae (Rosado-Lugo et al 2022).…”

Section: Discussionmentioning

confidence: 99%

“…More recently, two molecules, TXA6101 and TXY6129, with substituted 2,6-difluorobenzamide scaffold, have been shown to inhibit the polymerization of both E. coli and Klebsiella pneumoniae FtsZ. Moreover, despite being substrates for efflux pumps, TXA6101 induced morphological changes in K. pneumoniae (Rosado-Lugo et al 2022). Studies in the past on the effects of PC190723 on E. coli have been confusing, with a few reports suggesting an effect on FtsZ polymerization resulting in cell filamentation (Kaul et al 2014), while others did not find any effect on EcFtsZ (Andreu et al 2010; Anderson et al 2012; Khare et al 2019).…”

Section: Discussionmentioning

confidence: 99%

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A salt bridge-mediated resistance mechanism to FtsZ inhibitor PC190723 revealed by a cell-based screen

Sharma

Poddar

Chakraborty

et al. 2022

Preprint

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Bacterial cell division proteins, especially the tubulin homolog FtsZ, have emerged as strong targets for developing new antibiotics. Several assays have been designed to screen for small molecules targeting FtsZ but rely upon a multitude of steps to validate the target and to ensure minimum toxicity to the eukaryotic cells. Here, we have utilized the fission yeast heterologous expression system to develop a single step cell-based assay to screen for small molecules that directly and specifically target the bacterial cell division protein FtsZ and are non-toxic to eukaryotic cells. As a proof-of-concept of the utility of this assay, we demonstrate the effect of the inhibitors sanguinarine, berberine and PC190723 on FtsZ. Though sanguinarine and berberine affect FtsZ polymerization, they exert a toxic effect on the cells. Further, using this assay system, we show that PC190723 affects Helicobacter pylori FtsZ function and gain new insights into the molecular determinants of resistance to PC190723. Based on sequence and structural analysis and site-specific mutations, we demonstrate that the presence of salt-bridge interactions between the central H7 helix and beta-sheet S10 and S7 mediate resistance to PC190723 in FtsZ. The single step in vivo cell-based assay using fission yeast enabled us to dissect the contribution of sequence-specific features of FtsZ and cell permeability effects associated with bacterial cell envelopes. Thus, our assay functions as a powerful tool to rapidly identify new molecules that specifically target the bacterial cell division protein FtsZ, or other polymeric bacterial cytoskeletal proteins, understand how they affect polymerization dynamics and study resistance determinants in targets.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A salt bridge-mediated resistance mechanism to FtsZ inhibitor PC190723 revealed by a cell-based screen

Sharma

Poddar

Chakraborty

et al. 2022

Preprint

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“…From the crystal structures of TXA707 and TXA6101 bound to FtsZ [ 162 ], it is interesting to note that TXA6101 shows structural flexibility compared to TXA707. This enables it to target the FtsZ from gram-negative bacteria as well, broadening the specificity of these compounds to previously less targeted class of bacteria with respect to anti-FtsZ therapy [ 163 ]. TXA6101 and TXA6129 ( Figure 7 and Table 1 ) show activity against Enterobacteriaceae family of pathogens when used in combination with efflux pump inhibitors [ 163 ].…”

Section: Challenges and Opportunitiesmentioning

confidence: 99%

Models versus pathogens: how conserved is the FtsZ in bacteria?

et al. 2023

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Combating anti-microbial resistance by developing alternative strategies is the need of the hour. Cell division, particularly FtsZ, is being extensively studied for its potential as an alternative target for anti-bacterial therapy. Bacillus subtilis and Escherichia coli are the two well-studied models for research on FtsZ, the leader protein of the cell division machinery. As representatives of gram-positive and gram-negative bacteria respectively, these organisms have provided an extensive outlook into the process of cell division in rod-shaped bacteria. However, research on other shapes of bacteria, like cocci and ovococci, lags behind that of model rods. Even though most regions of FtsZ show sequence and structural conservation throughout bacteria, the differences in FtsZ functioning and interacting partners establishes several different modes of division in different bacteria. In this review, we compare the features of FtsZ and cell division in the model rods B. subtilis and E. coli and the four pathogens - Staphylococcus aureus, Streptococcus pneumoniae, Mycobacterium tuberculosis, and Pseudomonas aeruginosa. Reviewing several recent articles on these pathogenic bacteria, we have highlighted the functioning of FtsZ, the unique roles of FtsZ-associated proteins, and the cell division processes in them. Further, we provide a detailed look at the anti-FtsZ compounds discovered and their target bacteria, emphasizing the need for elucidation of the anti-FtsZ mechanism of action in different bacteria. Current challenges and opportunities in the ongoing journey of identifying potent anti-FtsZ drugs have also been described.

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“…Although this cleft is thought to open as well in T-state EcFtsZ filaments [ 26 ], two charged residue pairs have been proposed to form salt bridges between helix H7 and the C-terminal beta sheet, preventing cleft access to PC190723 in resistant species [ 75 ]. An allosteric modulator equivalent to PC190723 is thus missing for E. coli FtsZ, in addition to the general problem in crossing the outer membrane of Gram-negative bacteria [ 76 ], although the analog TXA6101 is active on Klebsiella pneumoniae [ 77 ].…”

Section: The Ftsz Interdomain Cleftmentioning

confidence: 99%

The Search for Antibacterial Inhibitors Targeting Cell Division Protein FtsZ at Its Nucleotide and Allosteric Binding Sites

et al. 2022

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The global spread of bacterial antimicrobial resistance is associated to millions of deaths from bacterial infections per year, many of which were previously treatable. This, combined with slow antibiotic deployment, has created an urgent need for developing new antibiotics. A still clinically unexploited mode of action consists in suppressing bacterial cell division. FtsZ, an assembling GTPase, is the key protein organizing division in most bacteria and an attractive target for antibiotic discovery. Nevertheless, developing effective antibacterial inhibitors targeting FtsZ has proven challenging. Here we review our decade-long multidisciplinary research on small molecule inhibitors of bacterial division, in the context of global efforts to discover FtsZ-targeting antibiotics. We focus on methods to characterize synthetic inhibitors that either replace bound GTP from the FtsZ nucleotide binding pocket conserved across diverse bacteria or selectively bind into the allosteric site at the interdomain cleft of FtsZ from Bacillus subtilis and the pathogen Staphylococcus aureus. These approaches include phenotype screening combined with fluorescence polarization screens for ligands binding into each site, followed by detailed cytological profiling, and biochemical and structural studies. The results are analyzed to design an optimized workflow to identify effective FtsZ inhibitors, and new approaches for the discovery of FtsZ-targeting antibiotics are discussed.

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Evaluation of 2,6-difluoro-3-(oxazol-2-ylmethoxy)benzamide chemotypes as Gram-negative FtsZ inhibitors

Cited by 12 publications

References 27 publications

A salt bridge-mediated resistance mechanism to FtsZ inhibitor PC190723 revealed by a cell-based screen

A salt bridge-mediated resistance mechanism to FtsZ inhibitor PC190723 revealed by a cell-based screen

Models versus pathogens: how conserved is the FtsZ in bacteria?

The Search for Antibacterial Inhibitors Targeting Cell Division Protein FtsZ at Its Nucleotide and Allosteric Binding Sites

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