title: Cyclobenzaprine Raises ROS Levels in Leishmania infantum and Reduces Parasite Burden in Infected Mice.
abstract: The leishmanicidal action of tricyclic antidepressants has been studied and evidences have pointed that their action is linked to inhibition of trypanothione reductase, a key enzyme in the redox metabolism of pathogenic trypanosomes. Cyclobenzaprine (CBP) is a tricyclic structurally related to the antidepressant amitriptyline, differing only by the presence of a double bond in the central ring. This paper describes the effect of CBP in experimental visceral leishmaniasis, its inhibitory effect in trypanothione reductase and the potential immunomodulatory activity.
  
  
  
  In vitro antileishmanial activity was determined in promastigotes and in L. infantum-infected macrophages. For in vivo studies, L. infantum-infected BALB/c mice were treated with CBP by oral gavage for five days and the parasite load was estimated. Trypanothione reductase activity was assessed in the soluble fraction of promastigotes of L. infantum. For evaluation of cytokines, L. infantum-infected macrophages were co-cultured with BALB/c splenocytes and treated with CBP for 48 h. The supernatant was analyzed for IL-6, IL-10, MCP-1, IFN-γ and TNF-α. CBP demonstrated an IC50 of 14.5±1.1μM and an IC90 of 74.5±1.2 μM in promastigotes and an IC50 of 12.6±1.05 μM and an IC90 of 28.7±1.3 μM in intracellular amastigotes. CBP also reduced the parasite load in L. infantum-infected mice by 40.4±10.3% and 66.7±10.5% in spleen at 24.64 and 49.28 mg/kg, respectively and by 85.6±5.0 and 89.3±4.8% in liver at 24.64 and 49.28mg/kg, after a short-term treatment. CBP inhibited the trypanothione reductase activity with a Ki of 86 ± 7.7 μM and increased the ROS production in promastigotes. CBP inhibited in 53% the production of IL-6 in infected macrophages co-culture.
  
  
  
  To the best of our knowledge, this study is the first report of the in vivo antileishmanial activity of the FDA-approved drug CBP. Modulation of immune response and induction of oxidative stress in parasite seem to contribute to this efficacy.
authors.affiliation: Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil.
authors.forename: Edézio Ferreira
authors.initials: EF
authors.lastname: Cunha-Júnior
authors.affiliation: Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil.
authors.forename: Valter Viana
authors.initials: VV
authors.lastname: Andrade-Neto
authors.affiliation: Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, São Paulo, Brazil.|Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Marta Lopes
authors.initials: ML
authors.lastname: Lima
authors.affiliation: Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Thais Alves
authors.initials: TA
authors.lastname: da Costa-Silva
authors.affiliation: Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, São Paulo, Brazil.
authors.forename: Andres J
authors.initials: AJ
authors.lastname: Galisteo Junior
authors.affiliation: Centro de Investigaciones Biológicas (CSIC), Unidad Asociada Interacciones, Metabolismo y Bioanálisis CSIC-CEU, Madrid, Spain.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Abengózar
authors.affiliation: Center for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad CEU San Pablo, Madrid, Spain.
authors.forename: Coral
authors.initials: C
authors.lastname: Barbas
authors.affiliation: Centro de Investigaciones Biológicas (CSIC), Unidad Asociada Interacciones, Metabolismo y Bioanálisis CSIC-CEU, Madrid, Spain.
authors.forename: Luis
authors.initials: L
authors.lastname: Rivas
authors.affiliation: Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil.
authors.forename: Elmo Eduardo
authors.initials: EE
authors.lastname: Almeida-Amaral
authors.affiliation: Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Andre Gustavo
authors.initials: AG
authors.lastname: Tempone
authors.affiliation: Laboratório de Bioquímica de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil.
authors.forename: Eduardo Caio
authors.identifier: 0000-0003-2240-4519
authors.initials: EC
authors.lastname: Torres-Santos
mesh_terms: D000639:Amitriptyline / Q000008:administration & dosage / Q000031:analogs & derivatives*; D000818:Animals; D000981:Antiprotozoal Agents / Q000008:administration & dosage*; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D016753:Interleukin-10 / Q000235:genetics / Q000276:immunology; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology; D018314:Leishmania infantum / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D007898:Leishmaniasis, Visceral / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D059208:Parasite Load; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
chemical_list: D000981:Antiprotozoal Agents; D015850:Interleukin-6; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D000639:Amitriptyline; C004704:cyclobenzaprine; D007371:Interferon-gamma