Antileishmanial activity of crude extract and coumarin from Calophyllum brasiliense leaves against Leishmania amazonensis.

Infections by protozoans of the genus Leishmania are a major worldwide health problem, with high endemicity in developing countries. The drugs of choice for the treatment of leishmaniasis are the pentavalent antimonials, which show renal and cardiac toxicity. As part of a search for new drugs against leishmaniasis, we evaluated the in vitro leishmanicidal activity of the (-) mammea A/BB. The compound (-) mammea A/BB is a coumarin-type mammea purified from a dichloromethane crude extract of leaves of Calophyllum brasiliense Cambess (Clusiaceae). The isolated compound was characterized using spectral analyses by UV, infrared, nuclear magnetic resonance of (1)H, (13)C, distortionless enhancement by polarization transfer, correlation spectroscopy, heteronuclear multiple bond correlation, and heteronuclear multiple quantum coherence. The compound (-) mammea A/BB showed significant activity against promastigote and amastigote forms of L. amazonensis, with IC(50) (50% inhibition concentration of cell growth) at a concentration of 3.0 and 0.88 mug/ml and IC(90) (90% inhibition concentration of cell growth) of 5.0 and 2.3 microg/ml, respectively. The coumarin (-) mammea A/BB showed no cytotoxicity against J774G8 macrophages in culture, when it was tested at high concentrations that inhibited promastigote forms. Electron microscopy studies revealed considerable ultrastructural changes when promastigote forms of L. amazonensis were treated with 3.0 microg/ml of the coumarin (-) mammea A/BB for 72 h. We observed significant changes such as mitochondrial swelling with concentric membranes in the mitochondrial matrix and intense exocytic activity in the region of the flagellar pocket. Other alterations included the appearance of binucleate cells and multiple cytoplasmic vacuolization. These results showed that (-) mammea A/BB is a potent growth inhibitor of L. amazonensis and caused important changes in the parasite's ultrastructure. This study provided new perspectives on the development of novel drugs with leishmanicidal activity obtained from natural products.

Antileishmanial activity of Eugenol-rich essential oil from Ocimum gratissimum.

Leishmaniasis is a group of diseases with a large spectrum of clinical manifestations caused by protozoans of the genus Leishmania. Here we demonstrate the leishmanicidal activity of the essential oil of Ocimum gratissimum as well as its main constituent, eugenol. The eugenol-rich essential oil of O. gratissimum progressively inhibited Leishmania amazonensis growth at concentrations ranging from 100 to 1000 microg/ml. The IC50 (sub-inhibitory concentration) of the essential oil for promastigotes and amastigotes were respectively 135 and 100 microg/ml and the IC50 of eugenol was 80 microg/ml for promastigote forms. L. amazonensis exposed to essential oil at concentrations corresponding to IC50 for promastigotes and for amastigotes underwent considerable ultrastructural alterations, as shown by transmission electron microscopy. Two or more nuclei or flagella were observed in 31% and 23.3% of treated amastigote and promastigote forms, respectively, suggesting interference in cell division. Considerable mitochondrial swelling was observed in essential oil-treated promastigotes and amastigotes, which had the inner mitochondrial membrane altered, with a significant increase in the number of cristae; in some amastigotes the mitochondrial matrix became less electron-dense. The minimum inhibitory concentration for both promastigotes and amastigotes was 150 microg/ml. Pretreatment of mouse peritoneal macrophages with 100 and 150 microg/ml essential oil reduced the indices of association between promastigotes and the macrophages, followed by increased in nitric oxide production by the infected macrophages. The essential oil showed no cytototoxic effects against mammalian cells. This set of results suggests that O. gratissimum essential oil and its compounds could be used as sources for new antileishmanial drugs.

Avaliação farmacológica de princípios ativos presentes no chá de Erva-de-passarinho - Phoradendron latifolium (SW) Griseb

Nossos experimentos foram realizados com o intuito de avaliar informações populares que indicam que o chá de Erva-de-Passarinho é altamente vasoconstritor, assim como informações científicas referentes a outras espécies da mesma família que, segundo vários autores, apresentam efeitos hiper e hipotensores. O Phoradendron latifolium (SW) Griseb. por nós utilizado foi coletado em Maringá -PR, onde encontra-se parasitando a Figueira Branca entre outras árvores. Realizamos ensaios com o chá liofilizado de Erva-de-Passaronho sobre a motilidade do duodeno isolado de coelhos e sobre a pressão arterial de cães anestesiados com Nembutal. Observamos que o chá promoveu redução da motilidade duodenal até o máximo de 80% das contrações iniciais. Na Pressão Arterial o chá levou ao aparecimento de uma resposta bifásica, ocorrendo inicialmente uma hipertensão seguida de hipotenso que não foi bloqueada pela atropina ou propanolol. Na presença de cocaína foi suprimida a hipertensão. Podemos concluir que o efeito hipotensor não está relacionado com receptores muscarínicos ou adrenérgicos tipo e que o efeito hipertensor seja provavelmente devido a uma substância do tipo da tiramina.

Contribuição ao estudo fitoquímico do Phoradendron latífolíum (SW) Griseb. (Erva-de-passarinho)

Phoradendron latifolium (SW) Griseb. is one of many species of mistletoe growing wild in Brazil. The plant used in our experiments was gathered in Maringá-Paraná where it is found growing on the stacks and branches of different hosts.Two ethanolic extracts were prepared using dried and milled leaves. The solvent was evaporated in vacuum. The residue of the first extract was dissolved with propylene glicol, the other residue, after washed with chloroform was dissolved in the same vehicle. Both extracts showed hipotensive response when tested on nembutal anaesthetized rats. The phytochemical screening revealed the presence of volatile and non volatile acids, amino groups, steroids, phenols, gums, mucilage, condensed tannins and heterosidic anthraquinones, flavones and non-haemolytic saponines.

O Phoradendron latifolium (SW) Griseb. objeto de nossa pesquisa é um hemiparasita nativo da região de Maringá-PR onde é encontrado em vários hospedeiros. Utilizamos em nossos experimentos vegetais que tinham como hospedeiro a Figueira Branca (Ficus sp; Moraceae). Foram preparados extratos etanólicos das folhas estabilizadas a 45oC sendo que um deles foi evaporado a pressão reduzida e o resíduo dissolvido em propilenoglicol. O outro teve seu resíduo obtido após evaporação do etanol lavado com clorofórmio objetivando a eliminação de substâncias orgânicas de baixa popularidade, sendo também dissolvido em propilenoglicol. Observou-se que ambos extratos continham princípios ativos hipotensores quando testados na PA de ratos. A marcha fitoquímica evidenciou a presença de ácidos fixos e voláteis, amino grupos, esteróides, fenóis, gomas, mucilagens, taninos condensados, heterosídeos antraquinônicos, flavônicos e saponínicos não hemolíticos.