Synthesis and biological evaluation of phosphate isosters of fosmidomycin and analogs as inhibitors of Escherichia coli and Mycobacterium smegmatis 1-deoxyxylulose 5-phosphate reductoisomerases

Munier, Mathilde; Tritsch, Denis; Krebs, Fanny S.; Esque, Jérémy; Hemmerlin, Andréa; Rohmer, Michel; Stote, Roland H.; Grosdemange-Billiard, Catherine

doi:10.1016/j.bmc.2016.11.040

Cited by 10 publications

(16 citation statements)

References 36 publications

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“…Fosmidomycin is known to enter cells by the glycerol-3-phosphate transporter, GlpT [ 16 , 34 , 71 – 75 ]. However, studies of fosmidomycin entry mechanism are largely conducted in rich growth medium, where the effects of GlpT deletion are dramatic ( Fig 6A , [ 16 , 71 , 74 , 76 , 77 ]).…”

Section: Resultsmentioning

confidence: 99%

“…IspC is the first committed step of the MEP pathway, catalyzing the conversion of DXP to MEP. Fosmidomycin and analogs are potent, selective IspC inhibitors that show potent antimicrobial activity against many Gram-negative bacteria [ 16 , 25 , 29 , 33 , 34 ]. The antimicrobial effects of fosmidomycin have been studied extensively in rich growth medium, however, fosmidomycin activity is not well-studied in nutrient limitation conditions thought to be more relevant to the in vivo growth environments of pathogens during infection [ 35 – 37 ].…”

Section: Introductionmentioning

confidence: 99%

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Growth medium-dependent antimicrobial activity of early stage MEP pathway inhibitors

et al. 2018

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The in vivo microenvironment of bacterial pathogens is often characterized by nutrient limitation. Consequently, conventional rich in vitro culture conditions used widely to evaluate antibacterial agents are often poorly predictive of in vivo activity, especially for agents targeting metabolic pathways. In one such pathway, the methylerythritol phosphate (MEP) pathway, which is essential for production of isoprenoids in bacterial pathogens, relatively little is known about the influence of growth environment on antibacterial properties of inhibitors targeting enzymes in this pathway. The early steps of the MEP pathway are catalyzed by 1-deoxy-d-xylulose 5-phosphate (DXP) synthase and reductoisomerase (IspC). The in vitro antibacterial efficacy of the DXP synthase inhibitor butylacetylphosphonate (BAP) was recently reported to be strongly dependent upon growth medium, with high potency observed under nutrient limitation and exceedingly weak activity in nutrient-rich conditions. In contrast, the well-known IspC inhibitor fosmidomycin has potent antibacterial activity in nutrient-rich conditions, but to date, its efficacy had not been explored under more relevant nutrient-limited conditions. The goal of this work was to thoroughly characterize the effects of BAP and fosmidomycin on bacterial cells under varied growth conditions. In this work, we show that activities of both inhibitors, alone and in combination, are strongly dependent upon growth medium, with differences in cellular uptake contributing to variance in potency of both agents. Fosmidomycin is dissimilar to BAP in that it displays relatively weaker activity in nutrient-limited compared to nutrient-rich conditions. Interestingly, while it has been generally accepted that fosmidomycin activity depends upon expression of the GlpT transporter, our results indicate for the first time that fosmidomycin can enter cells by an alternative mechanism under nutrient limitation. Finally, we show that the potency and relationship of the BAP-fosmidomycin combination also depends upon the growth medium, revealing a striking loss of BAP-fosmidomycin synergy under nutrient limitation. This change in BAP-fosmidomycin relationship suggests a shift in the metabolic and/or regulatory networks surrounding DXP accompanying the change in growth medium, the understanding of which could significantly impact targeting strategies against this pathway. More generally, our findings emphasize the importance of considering physiologically relevant growth conditions for predicting the antibacterial potential MEP pathway inhibitors and for studies of their intracellular targets.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Growth medium-dependent antimicrobial activity of early stage MEP pathway inhibitors

et al. 2018

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“…In an attempt to improve the efficiency of such inhibitors, numerous analogues of fosmidomycin have been synthesized. From these results, it is clear that neither the retro-hydroxamate chelating moiety nor the phosphonate anchoring group can be replaced without a drastic loss of activity except for the inversion of the retrohydroxamate into a hydroxamate (compounds 4a,b , Scheme 1 ) [ 8 ] and for the replacement of a phosphonate with a phosphate group (compounds 5a,b and 6a,b , Scheme 1 ) as we previously reported [ 9 ].…”

Section: Introductionmentioning

confidence: 69%

“…The synthesis of the diethyl α,α-difluorophosphonate 14 was achieved by previously reported methods [ 21 ]. Formylation with the mixed acetyl/formyl anhydride generated in situ from a formic acid and acetic anhydride mixture led to the N -formylated compound 15a , which was obtained as a mixture of conformers due to the restricted rotation around the C-N bond [ 9 , 24 , 25 , 26 ] and the large dipole moment of the C-F bond [ 27 ]. Acetylation with a mixture of acetic anhydride and pyridine gave the N -acetylated analogue 15b as previously described [ 21 ].…”

Section: Resultsmentioning

confidence: 99%

“…Even if the phosphate derivatives (compounds 5a,b , Scheme 1 ) were previously shown to be more potent inhibitors of the Synechocystis DXR than their phosphonate analogues [ 10 ], this conclusion is not valid for all DXRs, as we have shown that they are less efficient against the E. coli and Mycobacterium smegmatis DXRs [ 9 ]. Although a phosphonate group is considered to be isosteric to a phosphate group, some differences such as the pKa values and the C-O-P/C-C-P bond angles might impact their binding in the enzyme active site.…”

Section: Introductionmentioning

confidence: 87%

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α,α-Difluorophosphonohydroxamic Acid Derivatives among the Best Antibacterial Fosmidomycin Analogues

Dreneau¹,

Krebs²,

Munier³

et al. 2021

Molecules

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Three α,α-difluorophosphonate derivatives of fosmidomycin were synthesized from diethyl 1,1-difluorobut-3-enylphosphonate and were evaluated on Escherichia coli. Two of them are among the best 1-deoxy-d-xylulose 5-phosphate reductoisomerase inhibitors, with IC50 in the nM range, much better than fosmidomycin, the reference compound. They also showed an enhanced antimicrobial activity against E. coli on Petri dishes in comparison with the corresponding phosphates and the non-fluorinated phosphonate.

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Recent Insights Into Mechanism and Structure of MEP Pathway Enzymes and Implications for Inhibition Strategies

DeColli

Johnston

Meyers

2020

Comprehensive Natural Products III

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Synthesis and biological evaluation of phosphate isosters of fosmidomycin and analogs as inhibitors of Escherichia coli and Mycobacterium smegmatis 1-deoxyxylulose 5-phosphate reductoisomerases

Cited by 10 publications

References 36 publications

Growth medium-dependent antimicrobial activity of early stage MEP pathway inhibitors

Growth medium-dependent antimicrobial activity of early stage MEP pathway inhibitors

α,α-Difluorophosphonohydroxamic Acid Derivatives among the Best Antibacterial Fosmidomycin Analogues

Recent Insights Into Mechanism and Structure of MEP Pathway Enzymes and Implications for Inhibition Strategies

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