5-Nitrothiazol-Clofibric Acid Analogs as Potent Agents Against Nitazoxanide-Resistant Giardia lamblia: Modulation of Glycolytic Genes and Molecular Dynamics Studies.

Beatriz Hernández-Ochoa, Blanca Colin-Lozano, Gabriel Navarrete-Vázquez, Sergio Enríquez-Flores, Ignacio De la Mora-De la Mora, Verónica Pérez de la Cruz, Roberto Arreguin-Espinosa, Rosa Angélica Castillo-Rodríguez, José Antonio Velázquez-Aragón, Julieta Griselda Mendoza-Torreblanca, Noemí Cárdenas-Rodríguez, Daniel Ortega-Cuellar, Cindy Bandala, Abraham Vidal-Limon, Saúl Gómez-Manzo

Journal: Molecules (Basel, Switzerland) 2026;31(17):

PMID: 42738643

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Abstract

Giardiasis remains a significant global health issue, and its treatment is increasingly hampered by the emergence of drug resistance and cross-resistance between nitroimidazoles and nitazoxanide. In this study, we evaluated the in vitro antigiardial activity, cytotoxicity, and potential mechanisms of action of seven structural analogs (CLB 1-7) derived from a hybrid structure combining nitazoxanide and clofibric acid. Among them, compounds CLB-3 and CLB-5 emerged as the most potent candidates, demonstrating exceptional efficacy against a nitazoxanide-resistant (NTZr) strain of Giardia lamblia, with potencies up to 43 and 60 times higher than that of the reference drug, respectively. Furthermore, both compounds demonstrated high selectivity for the parasite, exhibiting low cytotoxicity in human Caco-2 cells. To investigate the mechanism of action, functional biochemical assays revealed that treatment with these derivatives significantly reduced the intracellular ATP levels and altered the ADP/ATP ratio, indicating a direct impact on the parasite's energy metabolism. These functional findings correlate closely with RT-qPCR transcriptional analyses and in silico molecular dynamics simulations; collectively, the data suggest a multi-target mechanism of action that may involve the disruption of energy homeostasis and the possible inhibition of key enzymes in the glycolytic pathway (PFOR, G6PD::6PGL, PK, and PPDK). In summary, these results identify compounds CLB-3 and CLB-5 as highly promising chemotherapeutic candidates for combating refractory giardiasis.

Address: Laboratorio de Investigación en Ciencias Ómicas y Epidemiología Microbiana, Hospital Infantil de México Federico Gómez, Secretaría de Salud, Mexico City 06720, Mexico.; Departamento de Química, Universidad La Salle, Mexico City 06140, Mexico.; Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Cuernavaca 62209, Morelos, Mexico.; Laboratorio de Biomoléculas y Salud Infantil, Instituto Nacional de Pediatría, Secretaría de Salud, Mexico City 04530, Mexico.; Neurobiochemistry and Behavior Laboratory, National Institute of Neurology and Neurosurgery "Manuel Velasco Suárez", Mexico City 14269, Mexico.; Departamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de México, México City 04510, Mexico.; Centro de Investigación en Ciencia Aplicada y Tecnología Avanzada (CICATA) Unidad Morelos, Instituto Politécnico Nacional, Xochitepec 62790, Morelos, Mexico.; Laboratorio de Oncología Experimental, Instituto Nacional de Pediatría, Mexico City 04530, Mexico.; Laboratorio de Neurociencias, Instituto Nacional de Pediatría, Secretaría de Salud, Mexico City 04530, Mexico.; Laboratorio de Nutrición Experimental, Instituto Nacional de Pediatría, Secretaría de Salud, Mexico City 04530, Mexico.; Laboratorio de Neurociencia Traslacional, Escuela Superior de Medicina, Instituto Politécnico Nacional, Mexico City 11340, Mexico.; Red de Estudios Moleculares Avanzados, Clúster Científico y Tecnológico BioMimic®, Instituto de Ecología A.C. (INECOL), Xalapa 91073, Veracruz, Mexico.; Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Secretaría de Salud, Mexico City 04530, Mexico.

DOI: 10.3390/molecules31172980

MeSH Terms: Giardia lamblia, Thiazoles, Humans, Nitro Compounds, Molecular Dynamics Simulation, Caco-2 Cells, Drug Resistance, Glycolysis, Antiprotozoal Agents, Molecular Docking Simulation