Short communication Antileishmanial activity of furoquinolines and coumarins from Helietta apiculata Maria Elena Ferreira a , Antonieta Rojas de Arias b , Gloria Yaluff a , Ninfa Vera de Bilbao a , Hector Nakayama a , Susana Torres a , Alicia Schinini a , Isabelle Guy c , Horacio Heinzen d , Alain Fournet e,Ã a Instituto de Investigaciones en Ciencias de la Salud, Department of Tropical Medicine, Asuncio ´n, Paraguay b Centro para el Desarrollo de la Investigacio ´n Cientı ´fica (CEDIC/FMB/Diaz Gill Medicina Laboratorial), Asuncio ´n, Paraguay c Substances d’Origine Naturelle et Analogues Structuraux, Faculte ´ de Pharmacie, Universite´ d’Angers, Angers, France d Ca ´tedra de Farmacognosia y Productos Naturales, Facultad de Quı ´mica, Montevideo, Uruguay e IRD US 084, Laboratoire de Pharmacognosie, Faculte ´ de Pharmacie, rue J.B. Cle´ment, 92296 Chˆ atenay-Malabry cedex, France article info Keywords: Helietta apiculata Rutaceae Furoquinolines Coumarins Leishmania amazonensis Oral administration abstract The bark infusion of H. apiculata are used to treat wound healing related to cutaneous leishmaniasis and as anti-inflammatory. Aim of the study: To isolate, purify active constituents of H. apiculata stem bark, and evaluate their in vitro and in vivo antileishmanial activities. Materials and methods: Isolation by chromatographic methods and chemical identification of furoquinoline alkaloids and coumarins, then evaluation of the in vitro leishmanicidal activity of these compounds against three strains of Leishmania sp. promastigotes and in vivo against Leishmania amazonensis in Balb/c mice. Results: Furoquinoline alkaloids and coumarins presented a moderate in vitro activity against promastigote forms of Leishmania sp. with IC 50 values in the range between 17 and 450 mg/ml. Balb/ c mice infected with Leishmania amazonensis were treated with g-fagarine by oral route, or with 3-(1’- dimethylallyl)-decursinol or (-)-heliettin by subscutaneous route for 14 days at 10mg/kg daily. In these conditions, g-fagarine, 3-(1’-dimethylallyl)-decursinol and (-)-heliettin showed the same efficacy as the reference drug reducing by 97.4, 95.6 and 98.6% the parasite loads in the lesion, respectively. Conclusion: These compounds showed significant efficacy in L. amazonensis infected mice, providing important knowledge to improve its potential role for a future use in the treatment of cutaneous leishmaniasis. & 2009 Elsevier GmbH. All rights reserved. Introduction Helietta apicultata Benth. (Rutaceae) is a South American tree or shrub growing in Paraguay, Brazil and Argentina commonly called ) yvyra ovi * in Paraguay or ) canela-de-veado * in Brazil. The infusion of bark of H. apiculata are used in Paraguay as wound healing that could be related to leishmanicidal ulcers, and also as anti-inflamma- tory when are associated with the hierba mate, Ilex paraguayensis St Hillaire (Aquafoliaceae). Following the evaluation of the leishmanici- dal activity of the quinolines (Fournet et al., 1993) we have found a common plant growing in Paraguay and containing analogs of these compounds, the furoquinoline alkaloids. In various studies (Goloub- kova et al. 1998) from Helietta apiculata collected in Argentina and Brazil, furoquinoline alkaloids and coumarins were isolated and identified. In a preliminary screening, alkaloidal extract of the stem bark of H. apiculata displayed an in vitro activity at a concentration of 50 mg/ml against three strains of promastigote forms of Leishmania species, L. braziliensis, L. amazonensis and L. donovani. Activity-directed fractiona- tion and purification of the alkaloidal and coumarins constituents of the stem bark gave five furoquinoline alkaloids and four coumarins identified by their physical and spectral data. In the present study, we describe the in vitro antileishmanial activity of the isolated compounds, and the efficacy of g-fagarine and two coumarins, (+)-3-(1’-dimethyllallyl)-decursinol and (-)-heliettin when they are administered by the oral route (g À fagarine) or by subcutaneous (coumarins) via in the Leishmania amazonensis infected Balb/c mice. Materials and methods General experimental procedures UV spectra were measured on a Shimadzu UV 1601 spectro- meter in MeOH. The 1 H-NMR and 13 C-NMR spectra were obtained ARTICLE IN PRESS Contents lists available at ScienceDirect journal homepage: www.elsevier.de/phymed Phytomedicine 0944-7113/$ - see front matter & 2009 Elsevier GmbH. All rights reserved. doi:10.1016/j.phymed.2009.09.009 Ã Corresponding author. Tel.: +33 0 1 46 83 59 69; fax: +33 0 1 46 83 53 99. E-mail address: alain.fournet@ird.fr (A. Fournet). Phytomedicine 17 (2010) 375–378