Milton N. da Silva

Antiplasmodial activity of the andiroba (Carapa guianensis Aubl., Meliaceae) oil and its limonoid-rich fraction.

ETHNOPHARMACOLOGICAL RELEVANCE From seeds of Carapa guianensis the Amazon native people extracts the andiroba oil, which is traditionally used as febrifuge, anti-malarial, insecticidal and repellant. The non-saponifiable fraction separated from the oil is rich in limonoids, which assigns its pharmacological effects. MATERIALS AND METHODS The andiroba oil and its limonoid-rich fraction were submitted to in vitro antiplasmodial bioassay using W(2) and Dd(2) strains of Plasmodium falciparum. The acute toxicity of andiroba oil was evaluated. The limonoid-rich fraction was subjected to fractionation and identified its major constituents. RESULTS Andiroba oil and its limonoid-rich fraction inhibited the growth of W(2) clone in 100%, between 24 and 72 h, at concentrations of 8.2 μg/mL and 3.1 μg/mL, respectively. Under the same conditions, the parasitaemia of Dd(2) clone provoked by the andiroba oil showed inhibition of 31% (IC(50) >82 μg/mL) with a time-dependent relationship of 24h and inhibition of 88% (IC(50) 8.4 μg/mL) after 72 h, while for the limonoid-rich fraction the inhibition of Dd(2) clone was 56% (IC(50) 2.8μg/mL) at 24h and 82% (IC(50) 0.4 μg/mL) after 72 h. Andiroba oil in acute toxicity test with a fixed dose (LD(50) >2000 mg/kg) was not toxic The limonoids identified in the oil were gedunin, 6α-acetoxygedunin, 7-deacetoxy-7-oxogedunin, 7-deacetylgedunin, 1,2-dihydro-3β-hydroxy-7-deacetoxy-7-oxogedunin and andirobin. Gedunin and derivatives has been reputed as anti-malarials. CONCLUSION The results support the traditional use of andiroba oil as antiplasmodial, which additionally proved not to be toxic in bioassays conducted with mice.

Potencial alelopático de catequinas de Tachigali myrmecophyla (leguminosae)

13 C NMR spectral data and comparison with literature data. The compounds (+)-catechin and epicatechin were submitted to germination inhibition and radicle and hypocotyl growth assays. Results showed some significant activities confirming the initial hypothesis about allelopathic properties of that plant.

Chemical constituents and allelopathic and antioxidant activities of Alchorneopsis floribunda Müll. Arg. (Euphorbiaceae)

Chemical investigation of extracts from the stems and leaves of Alchorneopsis floribunda Müll. Arg., collected in the Amazon region, was performed. The main isolated compounds were triterpenes (α-amyrin, β-amyrin, lupeol, betulin, betulinic acid, uvaol, erythrodiol and oleanolic acid) and phenolic acid derivatives from 4-hydroxybenzoic acid (gallic and protocatechuic acids and isocorilagin). In the germination assays, high inhibitory allelopathic effects of the extracts and isocorilagin were observed and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of isocorilagin was higher than those of the standards used (Trolox and butylated hydroxyanisole). This is the first chemical study of the genus Alchorneopsis (Euphorbiaceae).

Identification and antioxidant activity of several flavonoids of Inga edulis leaves

Um extrato metanol-agua das folhas de Inga edulis foi fracionado para identificar os compostos polifenolicos. Os compostos identificados foram o acido galico, a catequina, a epicatequina, a miricetina-3-ramnopiranosideo, a quercetina-3-glucopiranosideo e a quercetina-3-ramnopiranosideo. A capacidade antioxidante do extrato e dos polifenois puros foi medida pelo teste ORAC e comparada com o teor em fenolicos totais (TP). O extrato bruto seco apresentou valores de ORAC (11.16 mmol TE per g) e TP (496.5 mg GAE per g) muito altos. Os compostos identificados foram responsaveis, respectivamente, por 9.53 % e 12.10 % dos valores ORAC e de TP do extrato de folhas de Inga edulis.

Limonoids of the phragmalin type from Swietenia macrophylla and their chemotaxonomic significance.

Species of Swietenia elaborate limonoid chemistry along only one route, which leads to the mexicanolide type in most species and the phragmalin type in S. mahogani. A hexane extract from leaves of S. macrophylla afforded six new phragmalins with a 8,9,30-ortho-ester unit, namely, 6-O-acetylswietephragmin E (1), 3beta-O-destigloyl-3beta-O-benzoyl-6-O-acetylswietephragmin E (2), 12alpha-acetoxyswietephragmin C (3), 3beta-O-destigloyl-3beta-O-benzoyl-12alpha-acetoxyswietephragmin C (4), 12alpha-acetoxyswietephragmin D (5), and 3beta-O-destigloyl-3beta-O-benzoyl-12alpha-acetoxyswietephragmin D (6). This appears to be the first record of phragamalins from S. macrophylla, and this study shows the potential of tricyclic [3.3.1(2,10).1(1,4)]-decane limonoids as taxonomically useful chemical markers in the Meliaceae.

Plants used to treat snakebites in Santarém, western Pará, Brazil: An assessment of their effectiveness in inhibiting hemorrhagic activity induced by Bothrops jararaca venom

ETHNOPHARMACOLOGICAL RELEVANCE The poor distribution and limited availability of antivenoms in Brazil have led to greater use of plants to treat snakebites. Very often such plants are the only alternative available to riverside communities. MATERIALS AND METHODS Direct questionnaire-based interviews were conducted with members of the Cucurunã, São Pedro and Alter do Chão communities in Santarém, Pará, Brazil. For each of the 12 most frequently mentioned species aqueous extracts were prepared and the phytochemical profiles determined by thin layer chromatography. The concentrations of phenolic compounds (tannins and flavonoids) in the aqueous extracts were determined by colorimetric assays. To assess inhibition of the hemorrhagic activity of Bothrops jararaca venom, solutions containing the venom mixed with aqueous extracts in the ratios 1:12 and 1:48 were tested (w/w). SDS-PAGE and Western blot were used to assess the action of the extracts on Bothrops jararaca venom. RESULTS In all, 24 plants belonging to 19 families were mentioned in the survey as being used to treat snakebites. Leaves (84%), seeds (60.9%) and inner bark (53%) were cited as the most frequently used parts in folk remedies, which were usually prepared in the form of a decoction (62.5%), tincture (45%) or maceration (22.5%). Hemorrhage induced by Bothrops jararaca venom was completely inhibited by aqueous extracts of Bellucia dichotoma, Connarus favosus, Plathymenia reticulata and Philodendron megalophyllum, which had a high phenolic content and contained condensed and hydrolyzable tannins. The results of SDS-PAGE showed that some venom protein bands were not visible when the venom was preincubated with the extracts that had completely inhibited hemorrhagic activity of the venom. Western blot showed that the extracts did not have any enzymatic action on the proteins in the venom as it failed to detect low-molecular-weight bands, which are indicative of possible enzymatic cleavage. CONCLUSIONS Traditional use of plants to treat snakebites is a common practice in the western region of Pará, Brazil. Our findings show that some plant extracts were able to inhibit snake venom-induced hemorrhage in vitro. In vivo studies are being carried out to validate the traditional use of these species to treat snakebites.

Dihydroflavonols from the leaves of Derris urucu (Leguminosae): structural elucidation and DPPH radical-scavenging activity

Derris urucu e uma planta da Amazonia com propriedades inseticida e ictiotoxica. Estudos com esta especie reportam a presenca de flavonoides, principalmente rotenoides, bem como de estilbenos. A partir do extrato etanolico das folhas de Derris urucu (Leguminosae), tres novos diidroflavonoides, denominados urucuol A (1), B (2) e C (3) e o diidroflavonol isotirumalina (4), foram isolados e identificados. As estruturas destes compostos foram elucidadas por uma extensiva analise espectroscopica de RMN uni e bidimensional, UV, IV e dados de EM, alem de comparacao com dados da literatura. Os compostos isolados (1-4) foram avaliados quanto ao seu potencial sequestrador do radical DPPH• e apresentaram baixo poder antioxidante quando comparados ao antioxidante comercial trans-resveratrol.

Anti-Inflammatory and Antinociceptive Studies of Hydroalcoholic Extract from the Leaves of Phyllanthus brasiliensis (Aubl.) Poir. and Isolation of 5-O-β-d-Glucopyranosyljusticidin B and Six Other Lignans

The aim of this study was to investigate the chemical composition and the antiinflammatory/antinociceptive properties of the hydroalcoholic extract derived from the leaves of Phyllanthus brasiliensis (HEPB) in rodents. A new arylnaphthalene lignan glycoside, 5-O-β-d-glucopyranosyljusticidin B, together with six known lignans, were isolated from HEPB. 1D and 2D NMR experiments and HRMS were used to elucidate the structure of the new compound. HEPB toxicity and antinociceptive activity were evaluated through acute oral toxicity and formalin models in mice, respectively. The anti-inflammatory effects of HEPB were assessed using carrageenan- and dextran-induced paw edema models in rats. HEPB showed low toxicity. Oral administration of HEPB reduced paw edema induced by carrageenan, but not by dextran. HEPB and its fractions from FR6 to FR10 (FR6-10) inhibited the neurogenic and inflammatory phases of formalin-induced linking, demonstrating its antinociceptive activity. These results indicated that lignans from Phyllanthus brasiliensis exerted antinociceptive/anti-inflammatory effects not related to the histaminergic pathway.

Using LC and Hierarchical Cluster Analysis as Tools to Distinguish Timbó Collections into Two Deguelia Species: A Contribution to Chemotaxonomy

The species Deguelia utilis and Deguelia rufescens var. urucu, popularly known as “timbó,” have been used for many years as rotenone sources in insecticide formulations. In this work, a method was developed and validated using a high-performance liquid chromatography-photodiode array (HPLC-PDA) system, and results were analyzed using hierarchical cluster analysis (HCA). By quantifying the major rotenoids of these species, it was possible to establish a linear relation between them. The ratio between the concentrations of rotenone and deguelin for D. utilis is approximately 1:0.8, respectively, while for D. rufescens var. urucu it is 2:1. These results may help to distinguish these species contributing to their taxonomic identification.

Breve revisão etnobotânica, fitoquímica e farmacologia de Stryphnodendron adstringens utilizada na Amazônia

O Stryphnodendron adstringens (Mart.) e uma especie largamente distribuida pelas cinco regioes brasileiras, conhecida popularmente como: barbatimao, barba-de-timao, boraozinho-roxo, casca-da-virgindade, uabatimo, abaramotemo, casca-da-mocidade, faveiro e enche-cangalha. Na regiao norte, essa especie existe principalmente nos Estados do Acre, Amazonas, Amapa, Roraima e Para. Na medicina popular, Stryphnodendron adstringens (Mart.) e uma planta utilizada no tratamento de algumas patologias como: gonorreia, leucorreia, diarreia, ulceras, hemorragias vaginais, impinges, assim como, pode ser usada como agente anti-inflamatorio, cicatrizante, adstringente, hemostatico, antisseptico e anti-hipertensivo. O barbatimao possui varios compostos produzidos pelo seu metabolismo secundario, como: os alcaloides, terpenos, flavonoides, esteroides e taninos. e A atividade farmacologica dessa planta e devido principalmente ao teor de taninos presentes na especie. Dessa forma, esta revisao aborda uma breve descricao botânica, taxonomia e distribuicao geografica, assim como os aspectos etnobotânicos, fitoquimicos e farmacologicos da especie.