title: A lupane-triterpene isolated from Combretum leprosum Mart. fruit extracts that interferes with the intracellular development of Leishmania (L.) amazonensis in vitro.
abstract: BACKGROUND
  3beta,6beta,16beta-trihydroxylup-20(29)-ene is a lupane triterpene isolated from Combretum leprosum fruit. The lupane group has been extensively used in studies on anticancer effects; however, its possible activity against protozoa parasites is yet poorly known. The high toxicity of the compounds currently used in leishmaniasis chemotherapy stimulates the investigation of new molecules and drug targets for antileishmanial therapy.
  
  
  METHODS
  The activity of 3beta,6beta,16beta-trihydroxylup-20(29)-ene was evaluated against Leishmania (L.) amazonensis by determining the cytotoxicity of the compound on murine peritoneal macrophages, as well as its effects on parasite survival inside host cells. To evaluate the effect of this compound on intracellular amastigotes, cultures of infected macrophages were treated for 24, 48 and 96 h and the percentage of infected macrophages and the number of intracellular parasites was scored using light microscopy.
  
  
  RESULTS
  Lupane showed significant activity against the intracellular amastigotes of L. (L.) amazonensis. The treatment with 109 μM for 96 h reduced in 80 % the survival index of parasites in BALB/c peritoneal macrophages. At this concentration, the triterpene caused no cytotoxic effects against mouse peritoneal macrophages. Ultrastructural analyses of L. (L.) amazonensis intracellular amastigotes showed that lupane induced some morphological changes in parasites, such as cytosolic vacuolization, lipid body formation and mitochondrial swelling. Bioinformatic analyses through molecular docking suggest that this lupane has high-affinity binding with DNA topoisomerase.
  
  
  CONCLUSIONS
  Taken together, our results have showed that the lupane triterpene from C. leprosum interferes with L. (L.) amazonensis amastigote replication and survival inside vertebrate host cells and bioinformatics analyses strongly indicate that this molecule may be a potential inhibitor of topoisomerase IB. Moreover, this study opens major prospects for the development of novel chemotherapeutic agents with leishmanicidal activity.
authors.affiliation: Malaria and Leishmaniasis Bioassays platform, Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, Rondônia, Brazil. carolinateles@fiocruz.br.
authors.forename: Carolina Bioni Garcia
authors.initials: CB
authors.lastname: Teles
authors.affiliation: Center of Studies for Biomolecules Applied to Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, Rondônia, Brazil. leandroqmc@hotmail.com.
authors.forename: Leandro Soares
authors.initials: LS
authors.lastname: Moreira-Dill
authors.affiliation: Laboratory of Insect Bioecology, Universidade Federal de Rondônia (UNIR), Porto Velho, Rondônia, Brazil. alealsil@unir.br.|Laboratory of Medical Entomology, Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, Rondônia, Brazil. alealsil@unir.br.
authors.forename: Alexandre de Almeida
authors.initials: Ade A
authors.lastname: Silva
authors.affiliation: Research Laboratory of Chemistry of Natural Products, Universidade Federal de Rondônia (UNIR), Porto Velho, Rondônia, Brazil. vfacundo@unir.br.
authors.forename: Valdir Alves
authors.initials: VA
authors.lastname: Facundo
authors.affiliation: Structural Biochemistry Laboratory, PUC, Rio Grande do Sul, Porto Alegre, Brazil. walter@azevedolab.net.
authors.forename: Walter F
authors.initials: WF
authors.lastname: de Azevedo
authors.affiliation: Laboratory of Molecular Epidemiology, Research Institute for Tropical Diseases in Rondônia (IPEPATRO), Porto Velho, Rondônia, Brazil. isjcavalli@uesc.br.
authors.forename: Luiz Hildebrando Pereira
authors.initials: LH
authors.lastname: da Silva
authors.affiliation: Laboratory of Cellular Ultrastructure Hertha Meyer, Institute of Biophysics, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil. motta@biof.ufrj.br.
authors.forename: Maria Cristina M
authors.initials: MC
authors.lastname: Motta
authors.affiliation: Center of Studies for Biomolecules Applied to Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, Rondônia, Brazil. stabeli@fiocruz.br.
authors.forename: Rodrigo Guerino
authors.initials: RG
authors.lastname: Stábeli
authors.affiliation: Department of Biological Sciences, Universidade Estadual de Santa Cruz (UESC), Ilheús, Bahia, Brazil. isjcavalli@uesc.br.
authors.forename: Izaltina
authors.initials: I
authors.lastname: Silva-Jardim
mesh_terms: D000818:Animals; D029765:Combretum / Q000737:chemistry*; D003593:Cytoplasm / Q000469:parasitology; D004264:DNA Topoisomerases, Type I / Q000187:drug effects; D005260:Female; D005638:Fruit / Q000737:chemistry; D066298:In Vitro Techniques; D007894:Leishmania mexicana / Q000187:drug effects*; D007896:Leishmaniasis / Q000188:drug therapy / Q000469:parasitology*; D017737:Macrophages, Peritoneal / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D014315:Triterpenes / Q000302:isolation & purification / Q000494:pharmacology*; D053978:Pentacyclic Triterpenes
chemical_list: D004264:DNA Topoisomerases, Type I; D010936:Plant Extracts; D014315:Triterpenes; C480546:lupane; D053978:Pentacyclic Triterpenes