title: In vitro and experimental therapeutic studies of the calcium channel blocker bepridil: detection of viable Leishmania (L.) chagasi by real-time PCR.
abstract: The need for novel and efficacious drugs against neglected parasitic diseases, such as Leishmaniasis and American Trypanosomiasis, is certainly apparent. In this work, we evaluated the in vitro potential of the calcium channel blocker bepridil against Leishmania spp. and Trypanosoma cruzi parasites and exploited an experimental assay using a hamster model with Leishmania (L.) chagasi, with a real-time PCR method for therapeutic evaluation. Bepridil was in vitro effective against promastigotes and intracellular amastigotes of L. (L.) chagasi, with 50% inhibitory concentration (IC(50)) values of 3.81 and 21.55 μM, respectively. Leishmania (L.) amazonensis, L. (L.) major and L. (V.) braziliensis promastigotes and T. cruzi trypomastigotes were also susceptible to bepridil, with in vitro selectivity toward parasites and IC(50) values in the range of 3 to 7 μM. The mammalian cytotoxicity using LLC-MK2 cells resulted in an IC(50) value of 62.67 μM. However, bepridil showed lack of activity at 12 mg/kg in the experimental hamster model infected with L. (L.) chagasi parasites. However, the real-time PCR was a promising tool for the accurate and fast quantification of RNA of living parasites in the liver and spleen of infected hamsters after treatment, eliminating time-consuming light microscopy evaluations.
authors.affiliation: Laboratory of Applied Toxinology on Anti-Parasitic Drugs, Department of Parasitology, Instituto Adolfo Lutz Avenida Dr. Arnaldo, 351, 8 andar. Cerqueira César, CEP 01246-902 São Paulo, SP, Brazil.
authors.forename: Juliana Q
authors.initials: JQ
authors.lastname: Reimão
authors.forename: Fábio A
authors.initials: FA
authors.lastname: Colombo
authors.forename: Vera L
authors.initials: VL
authors.lastname: Pereira-Chioccola
authors.forename: André G
authors.initials: AG
authors.lastname: Tempone
mesh_terms: D000818:Animals; D015764:Bepridil / Q000494:pharmacology* / Q000627:therapeutic use; D002121:Calcium Channel Blockers / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003199:Computer Systems; D006224:Cricetinae; D004195:Disease Models, Animal; D005260:Female; D020128:Inhibitory Concentration 50; D007891:Leishmania / Q000145:classification / Q000187:drug effects; D018314:Leishmania infantum / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000188:drug therapy*; D008253:Macaca mulatta; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000469:parasitology; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D016053:RNA, Protozoan / Q000302:isolation & purification; D014349:Trypanosoma cruzi / Q000187:drug effects
chemical_list: D002121:Calcium Channel Blockers; D016053:RNA, Protozoan; D015764:Bepridil