Phytomedicine 19 (2012) 413–417 Contents lists available at SciVerse ScienceDirect Phytomedicine j ourna l ho mepage: www.elsevier.de/phymed Anti-leishmanial activity of alkaloidal extracts obtained from different organs of Aspidosperma ramiflorum Ananda de Castro Cunha a , Talita Perez Cantuaria Chierrito a , Gerzia Maria de Carvalho Machado b , Leonor Laura Pinto Leon b , Cleuza Conceic ¸ ão da Silva c , Julio Cesar Tanaka c , Lauro Mera de Souza d , Regina Aparecida Correia Gonc ¸ alves a , Arildo José Braz de Oliveira a,∗ a Departamento de Farmácia, Universidade Estadual de Maringá, Avenida Colombo, 5790, CEP 87.020-900, Maringá, PR, Brazil b Departamento de Imunologia, Instituto Oswaldo Cruz - Fiocruz, Rio de Janeiro, RJ, Brazil c Departamento de Química, Universidade Estadual de Maringá, Maringá, PR, Brazil d Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR, Brazil a r t i c l e i n f o Keywords: Leishmania Aspidosperma ramiflorum Monoterpenoid indole alkaloids a b s t r a c t The present study was designated to evaluate semi-quantitative antileishmanial activity of alkaloidal extracts that were obtained from 1 g of different parts of Aspidosperma ramiflorum (leaves, roots, seeds, and stem barks). Alkaloidal extracts of barks and leaves presented a good activity against the extracel- lular form (promastigotes) of Leishmania (L.) amazonensis. It is known that compounds responsible for the antileishmanial activity in the alkaloidal extracts from A. ramiflorum are the monoterpenoid indole alkaloids ramiflorine A and ramiflorine B, therefore extracts obtained from different plant parts were analyzed by electrospray ionization mass spectrometry (ESI-MS) in order to evidence the presence of these bioactive alkaloids. Based on these findings, alkaloidal extract from leaves was fractionated on preparative thin-layer chromatography in a bioassay-guided fractionation affording individual purified ramiflorines A and B. Both ramiflorines A and B showed significant activity against Leishmania (L.) ama- zonensis (LD 50 values of 18.5 ± 6.5 g/ml and 12.63 ± 5.52 g/ml, respectively). Our results are showing that alkaloidal extract from leaves is a promising alternative to the use of stem barks from A. ramiflorum. © 2011 Elsevier GmbH. All rights reserved. Introduction Leishmaniasis is a vector borne disease caused by an intra- macrophage protozoa, Leishmania (Order: Kinetoplastidae, Family: Trypanosomatidae, Genus: Leishmania), that is generally trans- mitted by sandflies, either Phlebotomus (Old world) or Lutzomyia (New World). Leishmania are among the most diverse of human pathogens, in terms of both geographical distribution and variety of clinical manifestations. The disease endemicity extends to over 88 countries, the major group (n = 72) belonging to the developing world while 13 belongs to the category of least developed coun- tries; sadly, its public health impact remains grossly neglected. Globally, the population at risk is 350 million, overall prevalence being 12 million, 2 million new cases occur annually and the Disability Adjusted Life Years (DALY) burden is considered to be 860,000 men and 1.2 million women (WHO 2010). This protozoa cause a disease with different clinical forms, among them cutaneous, hyperergic, mucocutaneous, and anergic diffuse leishmaniasis (Leon et al. 1990). The disease is endemic in ∗ Corresponding author. Tel.: +55 44 3011 4871; fax: +55 44 3011 4999. E-mail address: ajboliveira@uem.br (A.J.B.d. Oliveira). some geographical areas of Brazil, where it constitutes a serious health problem (Lonardoni et al. 1999; Leon et al. 2002). Despite a few research achievements, first-line chemotherapy is still based on pentavalent antimonials developed more than 50 years ago, which are toxic and prone to drug resistance. In some trials, alternative pharmaceutical formulations have been used to reduce the toxicity of these drugs (Frezard et al. 2000). Second- line drugs, such Amphotericin B, are more toxic and Amphotericin B’s lipid formulation is too expensive for routine use in underde- veloped countries (Murray 2001). Thus, there is a strong need for safer, cheaper and effective treatments against leishmaniasis. In order to find new drugs against leishmaniasis, we have stud- ied alkaloidal extracts (AEs) of Brazilian plants (Ferreira et al. 2004) such as Aspidosperma ramiflorum Muell. Arg. (Apocynaceae), com- monly known as “guatambu”, a tree which grows from 12 m to 30 m in height and is native to the forests in Southeastern Brazil (Lorenzi 1992). The basic crude extract from stem barks of A. ramiflorum showed a good antileishmanial activity (Ferreira et al. 2004) which we attributed to the presence of indole alkaloids and soon after we described the fractionation, purification and isolation of alkaloids responsible for the activity against Leishmania (L.) amazonensis (Tanaka et al. 2007). Our results revealed that dimeric corynanthe 0944-7113/$ – see front matter © 2011 Elsevier GmbH. All rights reserved. doi:10.1016/j.phymed.2011.12.004