Antitrypanosomal activity and evaluation of the mechanism of action of dehydrodieugenol isolated from Nectandra leucantha (Lauraceae) and its methylated derivative against Trypanosoma cruzi.
BACKGROUND
From a previous screening of Brazilian biodiversity for antiprotozoal activity, the hexane extract from leaves of Nectandra leucantha (Nees & Mart.) (Lauraceae) demonstrated activity against Trypanosoma cruzi. Chromatographic separation of this extract afforded bioactive dehydrodieugenol (1). Furthermore, methylated derivative 2 (dehydrodieugenol dimethyl ether) was prepared and also tested against T. cruzi.


OBJECTIVE
To examine the therapeutical potential of compounds 1 and 2 against T. cruzi as well as to elucidate the mechanism of action of bioactive compound 1 against T. cruzi.


METHODS
Crude hexane extract from leaves was subjected to chromatographic steps to afford bioactive compound 1. In order to analyze the effect of additional methyl group in the antiparasitic activity of 1, derivative 2 was prepared (both are no pan-assay interference compounds - PAINS). These compounds were evaluated in vitro against T. cruzi (trypomastigote and amastigote forms) and analyzed for the potential effect in host cells through the production of nitric oxide and reactive oxygen species. Finally, the plasma membrane effect of the most potent compound 1 was investigated in T. cruzi trypomastigotes.


RESULTS
Compounds 1 and 2 displayed activity against amastigotes of T. cruzi. Although both compounds promoted activity against intracellular amastigotes, the production of nitric oxide and reactive oxygen species of host cells were unaltered, suggesting an antiparasitic activity other than host cell activation. Considering 1 the most effective compound against T. cruzi, the interference in the plasma membrane of the trypomastigotes was investigated using the fluorescent probe SYTOX®Green. After a short-term incubation, the fluidity and integrity of the plasma membrane was completely altered, suggesting it as a primary target for compound 1 in T. cruzi.


CONCLUSIONS
Compounds 1 and 2 selectively eliminated the intracellular parasites without host cell activation and could be important scaffolds for the search of new hit compounds.
Center of Natural Sciences and Humanities, Federal University of ABC, Santo Andre, São Paulo 09210-180, Brazil; Institute of Food Chemistry, Technische Universität Braunschweig, Braunschweig, 38106, Germany.
Simone S
SS
Grecco
Center of Parasitology and Mycology, Adolfo Lutz Institute, São Paulo 01246-902, Brazil.
Thais A
TA
Costa-Silva
Institute of Food Chemistry, Technische Universität Braunschweig, Braunschweig, 38106, Germany.
Gerold
G
Jerz
Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of São Paulo, Diadema, São Paulo 09972-270, Brazil.
Fernanda S
FS
de Sousa
Center of Natural Sciences and Humanities, Federal University of ABC, Santo Andre, São Paulo 09210-180, Brazil; Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of São Paulo, Diadema, São Paulo 09972-270, Brazil.
Geanne A
GA
Alves Conserva
Center of Parasitology and Mycology, Adolfo Lutz Institute, São Paulo 01246-902, Brazil.
Juliana T
JT
Mesquita
Center of Parasitology and Mycology, Adolfo Lutz Institute, São Paulo 01246-902, Brazil.
Mariana K
MK
Galuppo
Center of Parasitology and Mycology, Adolfo Lutz Institute, São Paulo 01246-902, Brazil.
Andre G
AG
Tempone
LabMol, Laboratory for Molecular Modeling and Drug Design, Faculty of Pharmacy, Federal University of Goias, Goiânia, Goiás 74605-170, Brazil.
Bruno J
BJ
Neves
LabMol, Laboratory for Molecular Modeling and Drug Design, Faculty of Pharmacy, Federal University of Goias, Goiânia, Goiás 74605-170, Brazil.
Carolina H
CH
Andrade
Center of Natural Sciences and Humanities, Federal University of ABC, Santo Andre, São Paulo 09210-180, Brazil.
Rodrigo L O R
RL
Cunha
Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of São Paulo, Diadema, São Paulo 09972-270, Brazil.
Miriam
M
Uemi
Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of São Paulo, Diadema, São Paulo 09972-270, Brazil.
Patricia
P
Sartorelli
Center of Natural Sciences and Humanities, Federal University of ABC, Santo Andre, São Paulo 09210-180, Brazil. Electronic address: joao.lago@ufabc.edu.br.
João Henrique G
JH
Lago
D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001938:Brazil; D014355:Chagas Disease / Q000188:drug therapy*; D005054:Eugenol / Q000627:therapeutic use*; D019770:Lauraceae / Q000737:chemistry*; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D018515:Plant Leaves / Q000737:chemistry; D014349:Trypanosoma cruzi / Q000187:drug effects*
D000981:Antiprotozoal Agents; D010936:Plant Extracts; D005054:Eugenol
Antitrypanosomal; Nectandra leucantha; Neolignans; Plasma membrane permeabilization