Inhibitors for the bacterial ectonucleotidase Lp1NTPDase from Legionella pneumophila

Amelie Fiene, Younis Baqi, Enas M. Malik, Patrice Newton, Wenjin Li, Sang Yong Lee, Elizabeth L. Hartland, Christa E. Müller

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Legionella pneumophila is an aerobic, Gram-negative bacterium of the genus Legionella, which constitutes the major causative agent of Legionnaires’ disease. Recently a nucleoside triphosphate diphosphohydrolase (NTPDase) from L. pneumophila was identified and termed Lp1NTPDase; it was found to be a structural and functional homolog of mammalian NTPDases catalyzing the hydrolysis of ATP to ADP and ADP to AMP. Its activity is believed to contribute to the virulence of Legionella pneumophila. Therefore Lp1NTPDase inhibitors are considered as novel antibacterial drugs. However, only weakly potent compounds are available so far. In the present study, a capillary electrophoresis (CE)-based enzyme assay for monitoring the Lp1NTPDase activity was established. The enzymatic reaction was performed in a test tube followed by separation of substrate and products by CE and subsequent quantification by UV analysis. After kinetic characterization of the enzyme, a series of 1-amino-4-ar(alk)ylamino-2-sulfoanthraquinone derivatives structurally related to the anthraquinone dye Reactive Blue 2, a non-selective ecto-NTPDase inhibitor, was investigated for inhibitory activity on Lp1NTPDase using the CE-based enzyme assay. Derivatives bearing a large lipophilic substituent (e.g., fused aromatic rings) in the 4-position of the 1-amino-2-sulfoanthraquinone showed the highest inhibitory activity. Compounds with IC50 values in the low micromolar range were identified. The most potent inhibitor was 1-amino-4-[phenanthrene-9-yl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (28, PSB-16131), with an IC50-value of 4.24 μM. It represents the most potent Lp1NTPDase inhibitor described to date. These findings may serve as a starting point for further optimization. Lp1NTPDase inhibition provides a novel approach for the (immuno)therapy of Legionella infections.

Original languageEnglish
Pages (from-to)4363-4371
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume24
Issue number18
DOIs
Publication statusPublished - Sep 15 2016

Fingerprint

Legionella pneumophila
Capillary electrophoresis
Capillary Electrophoresis
Legionella
Cibacron Blue F 3GA
Enzyme Assays
Nucleosides
Adenosine Diphosphate
Inhibitory Concentration 50
Assays
Bearings (structural)
Enzymes
Gram-Negative Aerobic Bacteria
Derivatives
Legionnaires' Disease
Anthraquinones
Adenosine Monophosphate
Virulence
Hydrolysis
Bacteria

Keywords

  • Anthraquinone
  • Antibacterial drug
  • Capillary electrophoresis
  • Ecto-5′-nucleotidase
  • Legionnaires’ disease
  • Lpg1905
  • NTPDase
  • Reactive Blue 2
  • Structure–activity relationships
  • Synthesis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Inhibitors for the bacterial ectonucleotidase Lp1NTPDase from Legionella pneumophila. / Fiene, Amelie; Baqi, Younis; Malik, Enas M.; Newton, Patrice; Li, Wenjin; Lee, Sang Yong; Hartland, Elizabeth L.; Müller, Christa E.

In: Bioorganic and Medicinal Chemistry, Vol. 24, No. 18, 15.09.2016, p. 4363-4371.

Research output: Contribution to journalArticle

Fiene, A, Baqi, Y, Malik, EM, Newton, P, Li, W, Lee, SY, Hartland, EL & Müller, CE 2016, 'Inhibitors for the bacterial ectonucleotidase Lp1NTPDase from Legionella pneumophila', Bioorganic and Medicinal Chemistry, vol. 24, no. 18, pp. 4363-4371. https://doi.org/10.1016/j.bmc.2016.07.027
Fiene, Amelie ; Baqi, Younis ; Malik, Enas M. ; Newton, Patrice ; Li, Wenjin ; Lee, Sang Yong ; Hartland, Elizabeth L. ; Müller, Christa E. / Inhibitors for the bacterial ectonucleotidase Lp1NTPDase from Legionella pneumophila. In: Bioorganic and Medicinal Chemistry. 2016 ; Vol. 24, No. 18. pp. 4363-4371.
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abstract = "Legionella pneumophila is an aerobic, Gram-negative bacterium of the genus Legionella, which constitutes the major causative agent of Legionnaires’ disease. Recently a nucleoside triphosphate diphosphohydrolase (NTPDase) from L. pneumophila was identified and termed Lp1NTPDase; it was found to be a structural and functional homolog of mammalian NTPDases catalyzing the hydrolysis of ATP to ADP and ADP to AMP. Its activity is believed to contribute to the virulence of Legionella pneumophila. Therefore Lp1NTPDase inhibitors are considered as novel antibacterial drugs. However, only weakly potent compounds are available so far. In the present study, a capillary electrophoresis (CE)-based enzyme assay for monitoring the Lp1NTPDase activity was established. The enzymatic reaction was performed in a test tube followed by separation of substrate and products by CE and subsequent quantification by UV analysis. After kinetic characterization of the enzyme, a series of 1-amino-4-ar(alk)ylamino-2-sulfoanthraquinone derivatives structurally related to the anthraquinone dye Reactive Blue 2, a non-selective ecto-NTPDase inhibitor, was investigated for inhibitory activity on Lp1NTPDase using the CE-based enzyme assay. Derivatives bearing a large lipophilic substituent (e.g., fused aromatic rings) in the 4-position of the 1-amino-2-sulfoanthraquinone showed the highest inhibitory activity. Compounds with IC50 values in the low micromolar range were identified. The most potent inhibitor was 1-amino-4-[phenanthrene-9-yl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (28, PSB-16131), with an IC50-value of 4.24 μM. It represents the most potent Lp1NTPDase inhibitor described to date. These findings may serve as a starting point for further optimization. Lp1NTPDase inhibition provides a novel approach for the (immuno)therapy of Legionella infections.",
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N2 - Legionella pneumophila is an aerobic, Gram-negative bacterium of the genus Legionella, which constitutes the major causative agent of Legionnaires’ disease. Recently a nucleoside triphosphate diphosphohydrolase (NTPDase) from L. pneumophila was identified and termed Lp1NTPDase; it was found to be a structural and functional homolog of mammalian NTPDases catalyzing the hydrolysis of ATP to ADP and ADP to AMP. Its activity is believed to contribute to the virulence of Legionella pneumophila. Therefore Lp1NTPDase inhibitors are considered as novel antibacterial drugs. However, only weakly potent compounds are available so far. In the present study, a capillary electrophoresis (CE)-based enzyme assay for monitoring the Lp1NTPDase activity was established. The enzymatic reaction was performed in a test tube followed by separation of substrate and products by CE and subsequent quantification by UV analysis. After kinetic characterization of the enzyme, a series of 1-amino-4-ar(alk)ylamino-2-sulfoanthraquinone derivatives structurally related to the anthraquinone dye Reactive Blue 2, a non-selective ecto-NTPDase inhibitor, was investigated for inhibitory activity on Lp1NTPDase using the CE-based enzyme assay. Derivatives bearing a large lipophilic substituent (e.g., fused aromatic rings) in the 4-position of the 1-amino-2-sulfoanthraquinone showed the highest inhibitory activity. Compounds with IC50 values in the low micromolar range were identified. The most potent inhibitor was 1-amino-4-[phenanthrene-9-yl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (28, PSB-16131), with an IC50-value of 4.24 μM. It represents the most potent Lp1NTPDase inhibitor described to date. These findings may serve as a starting point for further optimization. Lp1NTPDase inhibition provides a novel approach for the (immuno)therapy of Legionella infections.

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