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Altered drug influx/efflux and enhanced metabolic activity in triclabendazole-resistant liver flukes

Published online by Cambridge University Press:  16 June 2005

L. I. ALVAREZ ,
H. D. SOLANA ,
M. L. MOTTIER ,
G. L. VIRKEL ,
I. FAIRWEATHER  and
C. E. LANUSSE
L. I. ALVAREZ
Affiliation:
Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Campus Universitario, 7000, Tandil, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
H. D. SOLANA
Affiliation:
Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Campus Universitario, 7000, Tandil, Argentina Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CICPBA), Buenos Aires, Argentina
M. L. MOTTIER
Affiliation:
Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Campus Universitario, 7000, Tandil, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
G. L. VIRKEL
Affiliation:
Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Campus Universitario, 7000, Tandil, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
I. FAIRWEATHER
Affiliation:
School of Biology and Biochemistry, Medical Biology Centre, The Queen's University of Belfast, Belfast, Northern Ireland, BT9 7BL, UK
C. E. LANUSSE
Affiliation:
Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Campus Universitario, 7000, Tandil, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
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Abstract

Triclabendazole (TCBZ) is a halogenated benzimidazole compound that possesses high activity against immature and adult stages of the liver fluke, Fasciola hepatica. The intensive use of TCBZ in endemic areas of fascioliasis has resulted in the development of liver flukes resistant to this compound. TCBZ sulphoxide (TCBZSO) and TCBZ sulphone (TCBZSO2) are the main molecules recovered in the bloodstream of TCBZ-treated animals. In order to gain some insight into the possible mechanisms of resistance to TCBZ, the goals of the work described here were: to compare the ex vivo transtegumental diffusion of TCBZ parent drug and its sulpho-metabolites (TCBZSO and TCBZSO2) into TCBZ-susceptible and -resistant liver flukes; and to assess the comparative pattern of TCBZ biotransformation by TCBZ-susceptible and -resistant F. hepatica. For the tegumental diffusion studies, TCBZ-susceptible (Cullompton) and -resistant (Sligo) adult flukes collected from untreated infected sheep were incubated (15–180 min) in KRT buffer containing either TCBZ, TCBZSO or TCBZSO2 (5 nmol.ml−1). For the metabolism studies, microsomal fractions obtained from TCBZ-susceptible and -resistant flukes were incubated for 60 min with TCBZ (40 μM), and the amount of the formed metabolic product (TCBZSO) was measured. Drug/metabolite concentrations were quantified by HPLC. All the assayed TCBZ-related molecules penetrated through the tegument of both TCBZ-susceptible and -resistant flukes. However, significantly lower (approximately 50%) concentrations of TCBZ and TCBZSO were recovered within the TCBZ-resistant flukes compared to the TCBZ-susceptible ones over the 180 min incubation period. The rate of TCBZ sulphoxidative metabolism into TCBZSO was significantly higher (39%) in TCBZ-resistant flukes. The flavin-monooxigenase (FMO) enzyme system appears to be the main metabolic pathway involved in the formation of TCBZSO in both TCBZ-susceptible and -resistant flukes. The altered drug influx/efflux and enhanced metabolic capacity identified in TCBZ-resistant liver flukes may account for the development of resistance to TCBZ.

Keywords

Fasciola hepaticatriclabendazoleresistance mechanismsdrug diffusion and metabolism
Type
Research Article
Information
Parasitology , Volume 131 , Issue 4 , October 2005 , pp. 501 - 510
Copyright
© 2005 Cambridge University Press

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