ABSTRACT
BACKGROUND: Leishmaniasis comprises several infectious diseases caused by protozoa parasites of Leishmania genus. In recent years, there has been a growing interest in the therapeutic use of natural products to treat parasitic diseases. Among them Croton cajucara Benth. (Euphorbiaceae) is a plant found in the Amazonian region with a history of safe use in folk medicine. PURPOSE: The purpose of this study was to investigate the effects of clerodane diterpenes, trans-dehydrocrotonin (DCTN), trans-crotonin (CTN) and acetylaleuritolic acid (AAA) obtained from powdered bark of C. cajucara against promastigotes, axenic and intracellular amastigotes of Leishmania amazonensis. Furthermore, the effects of DCTN and CTN on the trypanotiona reductase enzyme were also investigated. The extraction of the terpenes was carried out as previously reported (Maciel et al., 1998; 2003). METHODS: The effect of the isolated compounds (DCTN, CTN and AAA) from the bark of C. cajucara was assessed in vitro against promastigotes, axenic amastigotes and intracellular amastigotes of L. amazonensis by counting of remaining parasites in a Neubauer chamber in comparison to pentamidine used as standard drug. The action of natural products on trypanothione reductase was assessed using soluble protein fraction of promastigotes. The assays were performed by incubation with HEPES, EDTA, NADPH and trypanothione disulfide to quantify the NAPH consumption by TryR. RESULTS: The results showed very high efficacy, especially of the diterpene DCTN, against promastigotes (IC50 = 6.30 ± 0.06 µg/ml) and axenic amastigotes (IC50 = 19.98 ± 0.05 µg/ml) of L. amazonenesis. The cytotoxic effect of the best active natural product was evaluated on mouse peritoneal infected macrophages (IC50 = 0.47 ± 0.03 µg/ml in 24 h of culture), and the treatment revealed that DCTN never reaches toxic concentrations while reducing the infection and, most importantly, with no toxicity (>100 µg/ml with 0% of macrophage kill) when compared to pentamidine (37.5 µg/ml with 100% of macrophage kill). Furthermore, all of the natural products assayed on the trypanothione reductase enzyme inhibited the enzyme activity compared to the control. CONCLUSION: Clerodane diterpenes from C. cajucara showed promising in vitro antileishmanial effects against L. amazonensis, specially the DCTN with no macrophage toxicity up to the assayed concentration. In addition, the action on trypanothione reductase enzyme revealed a possible mechanism of action.
Subject(s)
Antiprotozoal Agents/pharmacology , Croton/chemistry , Diterpenes, Clerodane/pharmacology , Leishmania/drug effects , NADH, NADPH Oxidoreductases/antagonists & inhibitors , Animals , Antiprotozoal Agents/isolation & purification , Diterpenes , Diterpenes, Clerodane/isolation & purification , Inhibitory Concentration 50 , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/parasitology , Medicine, Traditional , Mice , Mice, Inbred BALB C , Molecular Structure , Plant Bark/chemistry , TriterpenesABSTRACT
Chromatographic fractionation of the chloroform extracts from the stem bark and stems of Guettarda grazielae resulted in the isolation of a new iridoid (guettardodiol, 1) and the secoiridoid sarracenin (2), described for the first time in this genus. The structural elucidation of these compounds was based on spectroscopic analyses (IR, MS as well as 1-D and 2-D NMR experiments).
Subject(s)
Iridoids/isolation & purification , Plant Extracts/chemistry , Rubiaceae/chemistry , Brazil , Chemical Fractionation , Iridoids/analysis , Magnetic Resonance Spectroscopy , Mass Spectrometry , Molecular Structure , Spectrophotometry, InfraredABSTRACT
In an effort to find effective and affordable ways to control of Aedes aegypti L. (Diptera: Culicidae), the larvicidal activities of 94 extracts from ten plant species belonging to eight families [Guettarda grazielae and Spermacoce verticillata (Rubiaceae), Coccoloba mollis and Triplaris americana (Polygonaceae), Eschweilera ovata (Lecytidaceae), Merremia aegyptia (Convolvulaceae), Ouratea nitida (Ochnnaceae), Protium heptaphyllum (Burseraceae), Rourea doniana (Connaraceae), and Tovomita brevistaminea (Guttiferae)], widely found in the northeast of Brazil, were initially screened at 250 microg/mL. Nineteen of the 94 extracts from six plant species (C. mollis, G. grazielae, M. aegyptia, R. doniana, S. verticillata, and T. americana) showed significant activity (>75% mortality) against the fourth instar larvae. Some of these extracts showed 100% mosquito larvae mortality, with LD(50) values lower 200 microg/mL. When compared to synthetic insecticide Temephos (LD(50) 8.7 microg/mL), two of them, hexane (LD(50) 12.1 microg/mL) and ethyl acetate (LD(50) 51.6 microg/mL) extracts from stems of R. doniana and G. grazielae, respectively, were the most effective. The isolation and purification of these extracts are in progress and evaluation of these compounds will be needed to identify the active component.
