LEISHMANICIDAL ACTIVITY OF PYCNOPORUS SANGUINEUS 497 Copyright © 2006 John Wiley & Sons, Ltd. Phytother. Res. 20, 497–499 (2006) DOI: 10.1002/ptr Copyright © 2006 John Wiley & Sons, Ltd. Received 3 September 2005 Accepted 26 January 2006 Leishmanicidal Activity of Pycnoporus sanguineus Edwin Correa 1 , Diana Cardona 1 *, Winston Quiñones 1 , Fernando Torres 1 , Ana E. Franco 2 , Ivan D. Vélez 3 , Sara Robledo 3 and Fernando Echeverri 1 * 1 Grupo de Química Orgánica de Productos Naturales-SIU, Universidad de Antioquia, P.O. Box 1226, Medellín-Colombia 2 Instituto de Biología Universidad de Antioquia, P.O. Box 1226, Medellín-Colombia 3 Programa para el Control y Estudio de Enfermedades Tropicales-SIU, Universidad de Antioquia P.O. Box 1226, Medellin-Colombia In the search for antiparasite compounds from the Colombian flora, an active compound against Leishmania (Viannia) panamensis amastigotes was isolated from the fungi Pycnoporus sanguineus. The structural elucid- ation was achieved with spectroscopic methods ( 1 H and 13 C NMR and MS). This compound was identified as ergosterol 5,8-endoperoxide. Copyright © 2006 John Wiley & Sons, Ltd. Keywords: Pycnoporus sanguineus; ergosterol endoperoxide; leishmanicidal activity. * Correspondence to: Fernando Echeverri, Grupo de Química Orgánica de Productos Naturales-SIU, Universidad de Antioquia, P.O. Box 1226, Medellín-Colombia E-mail: echeveri@quimbaya.udea.edu.co, dpcardona@gmail.com Contract/grant sponsor: Colciencias (Colombia). Contract/grant sponsor: Universidad de Antioquia. (Fig. 1) (Table 1). After the bioassay, 250.0 g of fresh fruiting body was cut in small pieces and macerated for 24 h in acetone, filtered and evaporated to dryness under vacuum. A portion of the extract was partitioned with ethyl acetate, dried over sodium sulphate, filtered and evaporated, yielding 1.5 g of a deep red solid; after that it was evaluated in vitro against L. (V.) panamensis intracellular amastigotes. Then, the extract was puri- fied using silica gel 60 column chromatography (Merck) eluted with CH 2 Cl 2 :EtOAC (2:1 v/v), CH 2 Cl 2 :EtOAC (1:1) and CH 2 Cl 2 :EtOAC (1:2). Less polar fractions were combined and fractionated on a lipophilic Sephadex LH-20 (Sigma) column, eluting with hexane:CH 2 Cl 2 :MeOH (2:1:1) and collecting fractions of 10 mL; thus, four new fractions were collected. The second one showed three compounds on TLC (silica gel 60 F 254 Merck), from which, compound 1 was puri- fied as a white solid (25.0 mg) by column chromatogra- phy on silica gel 60 eluted with benzene: ethyl acetate (100:0, 96:4, 92:8, 88:12, 84:16, 80:20). This compound was the most abundant in the extract prepared with hexane. INTRODUCTION The genus Pycnoporus (Polyporaceae) is constituted by four species, Pycnoporus sanguineus (L. Fr.) Murr., P. cinnabarinus (Jacq. Fr.) Karst., P. puniceus (Fr.) Ryv. and P. coccineus (Fr.) Bondartsev. & Singer (Gilbertson and Ryvarden, 1987; Ryvarden and Johansen, 1980) and is characterized by an orange-red to cinnabar colour of the pileus surface and pores. The members of the genus are known as white rot fungi. From the pantro- pical specie Pycnoporus sanguineus, several triterpenes and intense coloured phenaxozine-3-one compounds were isolated previously (Yokohama and Natori, 1975; Achenbach and Blümm, 1991). These compounds dis- play a resemblance to the pterin nucleus, a component of the folic acid moiety which is a target in the research for new antiparasite drugs (Ouellettea et al., 2002; Cunningham and Beverley, 2001). Using this structural approach, a bioassay-guided fractionation of P. san- guineus was carried out to determine activity against amastigotes of Leishmania (Viannia) panamensis. MATERIALS AND METHODS Fungi. Pycnoporus sanguineus was collected in Medellín City in November 2003 and a voucher specimen is held in the Herbarium of Universidad de Antioquia (HUA) under number 118775. The fungi were identified by one of the authors (A.E.F.). Extraction and isolation. Initially, 41.4 g of dry fruiting body was ground, extracted with hexane, evaporated and then the antileishmanial activity was determined Figure 1. Bioguided isolation of ergosterol endoperoxide 1. PHYTOTHERAPY RESEARCH Phytother. Res. 20, 497–499 (2006) Published online 18 April 2006 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ptr.1890