Targeting chalcone binding sites in living Leishmania using a reversible fluorogenic benzochalcone probe

Autor: Ariane S. Batista, Suellen D.S. Oliveira, Sébastien Pomel, Pierre-Henri Commere, Valérie Mazan, Moses Lee, Philippe M. Loiseau, Bartira Rossi-Bergmann, Eric Prina, Romain Duval
Přispěvatelé: Prina, Eric, Universidade Federal do Estado do Rio de Janeiro (UNIRIO), University of Illinois [Chicago] (UIC), University of Illinois System, Université Paris Saclay (COmUE), Biomolécules : Conception, Isolement, Synthèse (BioCIS), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY), Cytométrie (plateforme) - Flow Cytometry (platform), Institut Pasteur [Paris] (IP), Laboratoire d'innovation moléculaire et applications (LIMA), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Georgia State University, University System of Georgia (USG), Parasitologie moléculaire et Signalisation / Molecular Parasitology and Signaling, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Mère et enfant en milieu tropical : pathogènes, système de santé et transition épidémiologique (MERIT - UMR_D 216), Institut de Recherche pour le Développement (IRD)-Université Paris Cité (UPCité), The authors wish to acknowledge the following contributors: Prof. G. Lewin (BioCIS, Université Paris-Sud 11) for the kind gift of chalcones 4, 5 and 7, Dr. P. Leal (Universidade Federal de Santa Catarina) for the generous gift of xanthoxyline, synthetic precursor of flavokawain A 2 and chalcone 3, Prof. K. Miranda and Prof. F. Gomez (Universidade Federal de Rio de Janeiro) for providing with anti-vacuolar-type proton pyrophosphatase antibodies and for stimulating discussions regarding ACC, Prof. R. Coutinho-Silva (Universidade Federal de Rio de Janeiro) for helping with the promastigote microscopy experiments, Dr J. Moreira (Owkin) for translating the CNPq scientic proposal into Portuguese. Grant from the Conselho Nacional de Desenvolvimento Cientifco e Tecnologico, CNPq (401897/2010-9 to R. D.) is acknowledged., Mère et enfant en milieu tropical : pathogènes, système de santé et transition épidémiologique (MERIT - UMR_D 261)
Rok vydání: 2022
Předmět:
Zdroj: Biomedicine and Pharmacotherapy
Biomedicine and Pharmacotherapy, 2022, 149, pp.112784. ⟨10.1016/j.biopha.2022.112784⟩
ISSN: 0753-3322
1950-6007
DOI: 10.1016/j.biopha.2022.112784
Popis: International audience; Chalcones (1,3-diphenyl-2-propen-1-ones) either natural or synthetic have a plethora of biological properties including antileishmanial activities, but their development as drugs is hampered by their largely unknown mechanisms of action. We demonstrate herein that our previously described benzochalcone fluorogenic probe (HAB) could be imaged by fluorescence microscopy in live Leishmania amazonensis promastigotes where it targeted the parasite acidocalcisomes, lysosomes and the mitochondrion. As in the live zebrafish model, HAB formed yellow-emitting fluorescent complexes when associated with biological targets in Leishmania. Further, we used HAB as a reversible probe to study the binding of a portfolio of diverse chalcones and analogues in live promastigotes, using a combination of competitive flow cytometry analysis and cell microscopy. This pharmacological evaluation suggested that the binding of HAB in promastigotes was representative of chalcone pharmacology in Leishmania, with certain exogenous chalcones exhibiting competitive inhibition (ca. 20-30%) towards HAB whereas non-chalconic inhibitors showed weak capacity (ca. 3-5%) to block the probe intracellular binding. However, this methodology was restricted by the strong toxicity of several competing chalcones at high concentration, in conjunction with the limited sensitivity of the HAB fluorophore. This advocates for further optimization of this undirect target detection strategy using pharmacophore-derived reversible fluorescent probes.
Databáze: OpenAIRE
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