Abstract
Schistosomiasis, caused by a blood fluke of the genus Schistosoma, afflicts over 230 million people worldwide. Treatment of the disease relies on just one drug, praziquantel. Cnicin (Cn) is the sesquiterpene lactone found in blessed thistle (Centaurea benedicta) that showed antiparasitic activities but has not been evaluated against Schistosoma. However, cnicin has poor water solubility, which may limit its antiparasitic activities. To overcome these restrictions, inclusion complexes with cyclodextrins may be used. In this work, we evaluated the in vitro and in vivo antischistosomal activities of cnicin and its complexes with β-cyclodextrin (βCD) and 2-hydroxypropyl-β-cyclodextrin (HPβCD) against Schistosoma mansoni. Cnicin were isolated from C. benedicta by chromatographic fractionation. Complexes formed by cnicin and βCD (Cn/βCD), as well as by cnicin and HPβCD (Cn/HPβCD), were prepared by coprecipitation and characterized. In vitro schistosomicidal assays were used to evaluate the effects of cnicin and its complexes on adult schistosomes, while the in vivo antischistosomal assays were evaluated by oral and intraperitoneal routes. Results showed that cnicin caused mortality and tegumental alterations in adult schistosomes in vitro, also showing in vivo efficacy after intraperitoneal administration. The oral treatment with cnicin or Cn/βCD showed no significant worm reductions in a mouse model of schistosomiasis. In contrast, Cn/HPβCD complex, when orally or intraperitoneally administered to S. mansoni-infected mice, decreased the total worm load, and markedly reduced the number of eggs, showing high in vivo antischistosomal effectiveness. Permeability studies, using Nile red, indicated that HPβCD complex may reach the tegument of adult schistosomes in vivo. These results demonstrated the antischistosomal potential of cnicin in preparations with HPβCD.
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Acknowledgments
The authors are grateful to the FAPEMIG (Grant numbers PPM 00296/16, APQ 03536-16), CNPq (Grant numbers 487221/2012-5, 311913/2017-2, 437418/2018-9), and FAPESP (Grant 2016/22488-3) for financial support, as well as to CAPES, PIBIC/CNPq/UFJF, and CNPq for fellowships. We are also grateful to Dr. Pedro L. Pinto for assistance with S. mansoni life cycle maintenance at the Adolfo Lutz Institute (São Paulo, SP, Brazil), as well as to Lorena Rodrigues Riani (NIPPAN), Lívia Mara Silva, and Carolina Gasparetto Silva (CENTRALBIO) for technical assistance in UFJF. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior- Brazil (CAPES).
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Lucas S. Queiroz: investigation, formal analysis, and writing—original draft. Everton Allan Ferreira: investigation and formal analysis. Ana C Mengarda: investigation and formal analysis. Ayla das C. Almeida: investigation, formal analysis, and writing—original draft. Priscila de F. Pinto: investigation and formal analysis. Elaine S. Coimbra: investigation and formal analysis. Josué de Moraes: conceptualization, investigation, formal analysis, writing—original draft, and funding acquisition. Ângelo M. L. Denadai: conceptualization, investigation, formal analysis, writing—original draft, and funding acquisition. Ademar A. da Silva Filho: conceptualization, investigation, formal analysis, writing—original draft, resources, project administration, and funding acquisition.
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All experiments were conducted in conformity with the Brazilian Law for Guidelines for Care and Use of Laboratory Animals (Law 11790/2008). The protocol for experimental design was approved by the Comissão de Ética no Uso de Animais (CEUA), Brazil (Protocols ≠ CEUA 031/2017 and ≠ CEUA 007/2018). Animal studies are reported in compliance with the ARRIVE guidelines.
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Queiroz, L.S., Ferreira, E.A., Mengarda, A.C. et al. In vitro and in vivo evaluation of cnicin from blessed thistle (Centaurea benedicta) and its inclusion complexes with cyclodextrins against Schistosoma mansoni. Parasitol Res 120, 1321–1333 (2021). https://doi.org/10.1007/s00436-020-06963-2
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DOI: https://doi.org/10.1007/s00436-020-06963-2