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RCB20, an experimental benzimidazole derivative, affects tubulin expression and induces gross anatomical changes in Taenia crassiceps cysticerci

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Abstract

Helminth β-tubulins are the targets of benzimidazole (BZM) carbamate compounds. The specificity of the interactions between such compounds and their in vivo targets depends on the presence of specific amino acid residues in the target molecules. To discover new and effective anthelmintic drugs, we used a medicinal chemistry approach to synthesize a series of BZM derivatives that exploited the BZM moiety as a template. We have previously found that one compound, 2-(trifluoromethyl)-1H-benzimidazole (RCB20), has better in vitro and in vivo activity than albendazole sulfoxide (ABZSO). In the present study, the effect of RCB20 and ABZSO treatment on expression of Taenia crassiceps cysticerci cytoskeletal proteins such as actin, myosin II, and tubulin isoforms was examined. The effects of RCB20 and ABZSO after 11 days treatment of the parasites was evaluated by light, confocal, and electron microscopy, and by immunochemistry and immunohistochemistry. The RCB20-induced effects were more rapid than the ABZSO-induced effects on the parasites. In the RCB20-treated parasites, we observed gross-structural damage at the whole parasite level, particularly in the inner tissues and flame cells. Changes in the expression patterns of the cytoskeletal proteins, as assessed by immunohistochemistry and immunoblotting, revealed that the most important drug-induced effect on the parasites was a reduction in the expression level of tyrosinated α-tubulins. Our research findings suggest that RCB20 treatment affected posttranslational modification of parasite α-tubulin molecules, which involved removal of the α-tubulin carboxy-terminal tyrosine.

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Abbreviations

ABZSO:

Albendazole sulfoxide

BZM:

Benzimidazole

RCB20:

2-(trifluoromethyl)-1H-benzimidazole derivative

CTT:

Carboxy-terminal tyrosine

DAPI:

Diamino-2-phenylindole

DMSO:

Dimethyl sulfoxide

F-actin:

Filamentous actin

G-actin:

Globular actin

FC:

Flame cells

HCM:

Heavy chain myosin

LSCM:

Laser scanning confocal microscopy

MT:

Microtubule

MAb:

Monoclonal antibody

RT:

Room temperature

PBS:

Phosphate-buffered saline solution

SEM:

Scanning electron microscopy

PTM:

Posttranslational modifications

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Acknowledgments

This research was funded by the following grants: IN201003, IN216107, IN201510, and IX200610 from the Dirección General de Asuntos del Personal Académico (DGAPA)-UNAM, and V43629-M 080093 from the Consejo Nacional de Ciencia y Tecnología. AMN received a postdoctoral fellowship from DGAPA-UNAM. Microscopic expertise was provided by Gabriel Ortiz from IFICE-UNAM. We also thank Laura Valverde-Islas and Carlos Ibarra for their technical help and advice during microscopic analysis of the parasites. Our thanks also go to M.L.I. Rafael Ibarra for his enthusiastic guidance on this manuscript.

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Authors and Affiliations

  1. Facultad de Medicina, Departamento de Microbiología y Parasitología, Universidad Nacional Autónoma de México, 04510, México, DF, Mexico

    Adrián Márquez-Navarro, América Pérez-Reyes, Olivia Reynoso-Ducoing & Javier R. Ambrosio

  2. Facultad de Medicina, Departamento de Biología Celular y Tisular, Universidad Nacional Autónoma de México, 04510, México, DF, Mexico

    Armando Zepeda-Rodríguez

  3. Facultad de Química, Departamento de Farmacia, Universidad Nacional Autónoma de México, México, DF, 04510, Mexico

    Alicia Hernández-Campos, Francisco Hernández-Luis & Rafael Castillo

  4. Unidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Instituto Mexicano del Seguro Social, 06720, México, DF, Mexico

    Lilian Yépez-Mulia

Authors
  1. Adrián Márquez-Navarro
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  2. América Pérez-Reyes
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  3. Armando Zepeda-Rodríguez
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  6. Francisco Hernández-Luis
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  8. Lilian Yépez-Mulia
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  9. Javier R. Ambrosio
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Correspondence to Javier R. Ambrosio.

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Márquez-Navarro, A., Pérez-Reyes, A., Zepeda-Rodríguez, A. et al. RCB20, an experimental benzimidazole derivative, affects tubulin expression and induces gross anatomical changes in Taenia crassiceps cysticerci. Parasitol Res 112, 2215–2226 (2013). https://doi.org/10.1007/s00436-013-3379-2

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