Absolute configuration reassignment of two chromanes from Peperomia obtusifolia (Piperaceae) using VCD and DFT calculations

Previous analysis of the ECD spectra of two prenylated benzopyrans isolated from Peperomia obtusifolia, by means of the helicity rule for the chromane chromophore, resulted in the incorrect assignment of their absolute configuration, (5) instead of (R) for a deduced P-helicity of the chromane ring for the (+)-enantiomers. This was discovered by the application of DFT calculations and VCD spectroscopy. Experimental and calculated (B3LYP/6-31G(d)) VCD and IR spectra were compared, and a definitive absolute configuration of (+)-1 and (+)-2 is reassigned directly in solution as (R). The assumption of equatorial positioning of bulky groups, shown here to be invalid for the title molecules, is the underlying cause of the previous incorrect assignment of absolute configuration. Moreover, TDDFT (B3LYP/6-311++G(2d,2p)//B3LYP/6-31G(d)) calculations of ECD spectra have shown that both P- and M-helicity of the heterocyclic ring, for a given absolute configuration, lead to the same sign for the (1)L(b) ECD band, thus bringing into question the validity of the empirical ECD helicity rule for chromane molecules. (C) 2010 Elsevier Ltd. All rights reserved.

Geranylation of benzoic acid derivatives by enzymatic extracts from Piper crassinervium (Piperaceae)

The ability to carry out geranylations on aromatic substrates using enzymatic extracts from the leaves of Piper crassinervium (Piperaceae) was evaluated. A literature analysis pointed out its importance as a source of prenylated bioactive molecules. The screening performed on aromatic acceptors (benzoic acids, phenols and phenylpropanoids) including geranyl diphosphate as prenyl donor, showed the biotransformation of the 3,4-dihydroxybenzoic acid by the crude extract, and the p-hydroxybenzoic acid by both the microsomal fraction and the crude extract, after treating leaves with glucose. The analysis of the products allowed the identification of C- and O-geranylated derivatives, and the protease (subtilisin and pepsin) inhibition performed on the O-geranylated compounds showed weak inhibition. Electrophoretic profiles indicated the presence of bands/spots among 56-58 kDa and pI 6-7, which are compatible with prenyltransferases. These findings show that P. crassinervium could be considered as a source of extracts with geranyltransferase activity to perform biotransformations on aromatic substrates. (C) 2010 Elsevier Ltd. All rights reserved.

Resolution and Absolute Configuration Assignment of a Natural Racemic Chromane from Peperomia obtusifolia (Piperaceae)

The resolution of the natural racemic chromane 3,4-dihydro-5-hydroxy-2,7-dimethyl-8-(3 ``-methyl-2 ``-butenyl)-2-(4`-methyl-1`,3`-pentadienyl)-2H-1-benzopyran-6-carboxylic acid (1) isolated from the leaves of Peperomia obtusifolia has been accomplished using stereoselective HPLC. The absolute coil figuration of the resolved enantiomers was determined by the analysis of optical rotations and CD spectra. The finding of a racemic mixture instead of an enantiomerically pure metabolite raises questions about the final steps in the biosynthesis of this class of natural products, suggesting that the intramolecular chromane ring formation step may not be enzymatically controlled at all in P. obtusifolia. Chirality 21:799-801, 2009. (C) 2008 Wiley-Liss, Inc.

Prenylated benzoic acid derivatives from Piper aduncum L. and P. hostmannianum C. DC. (Piperaceae)

The bioactivity-guided fractionation of the crude extracts from leaves of Brazilian species Piper aduncum and Piper hostmannianum by means of bioautography using the fungi Cladosporium cladosporioides and C. sphaerospermum afforded prenylated methyl benzoate, chromenes, and dihydrobenzopyran derivatives as antifungal compounds. The isolation and structural elucidation of a new compound methyl 4-hydroxy-3-(2`-hydroperoxy-3`-methyl-3`-butenyl) benzoate were performed by application of chromatographic techniques and spectroscopic analyses. (C) 2009 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.

Sequestration of prenylated benzoic acid and chromenes by Naupactus bipes (Coleoptera: Curculionidae) feeding on Piper gaudichaudianum (Piperaceae)

The curculionid beetle Naupactus bipes (Germar, 1824) (Coleoptera: Curculionidae: Brachycerinae) has shown feeding preference for leaves of Piper gaudichaudianum, demonstrating an unexpected specificity for an insect considered to be a generalist. The leaves of P. gaudichaudianum contain the prenylated chromenes gaudichaudianic acid (4, major compound) and its methyl ester (5) in addition to a chromene (3) lacking one prenyl residue. In addition to 4, roots contain the chromone methyl ester (1) and methyl taboganate (2, major compound). Feeding on roots, larvae of N. bipes sequester exclusively the root-specific compounds 1 and 2. Adult beetles sequester the leaf-specific chromenes 3 and 4, but were found to also contain compounds 1 and 2 that are absent in leaves. Therefore, it is suggested that 1 and 2 are sequestered by larvae and can be found in the body of adult insects after long-term storage. In addition, 3 and 4, the major compounds in leaves were found to be associated with the eggs.

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)). Piper solmsianum is a shrub from Southeast Brazil in which many biologically active compounds were identified. The aim of this work was to establish a cell suspension culture system for this species. With this in mind, petiole and leaf explants obtained from in vitro plantlets were cultured in the presence of different plant growth regulator combinations (IAA, NAA, 2,4-D and BA). Root and indirect shoot adventitious formation, detected by histological analysis, was observed. Besides the different combinations of plant growth regulators, light regime and the supplement of activated charcoal (1.5 mg.l(-1)) were tested for callus induction and growth. Cultures maintained in light, on a 0.2 mg.l(-1) 2,4-D and 2 mg.l(-1) BA supplemented medium, and in the absence of activated charcoal, showed the highest calli fresh matter increment. From a callus culture, cell suspension cultures were established and their growth and metabolite accumulation studied. The achieved results may be useful for further characterization of the activated secondary metabolites pathways in in vitro systems of P. solmsianum.

Radical Scavenging Capacity of Piper arboreum and Piper tuberculatum (Piperaceae)

In the scope of our ongoing researchers on antioxidant compounds, twenty four extracts and fractions obtained from Piper arboreum Aublet and Piper tuberculatum Jacq. (Piperaceae) were screened for radical scavenging capacity (RSC) by using DPPH colorimetric assay. The strongest activity was found in ethyl acetate fractions from the leaves of P. arboreum IC(50) = 5.70 mu g/mL) and P. tuberculatum IC(50) = 8.40 mu g/mL). Hydromethanol fractions of the leaves of P. tuberculatum and P. arboreum showed moderate RSC, with values of IC(50) (mu g/mL) of 11.9 and 19.2, respectively. Additionally, a brief phytochemical study of the ethyl acetate fraction of P. arboreum leaves affording quercetin (1) and quercitrin (2), two flavonols with antioxidant activity previously described in the literature.

Sesquiterpenes from Xylaria sp., an endophytic fungus associated with Piper aduncum (Piperaceae)

Two new presilphiperfolane sesquiterpenes, 1 and 2, were isolated from the ethyl acetate extract of Xylaria sp., obtained from the leaves of Piper aduncum, along with two known eremophilane sesquiterpenes, phaseolinone (3) and phomenone (4). Chemical structures of 1 and 2 were established by analysis of spectroscopic data. The four compounds were tested in vitro for antifungal and cytotoxicity activities using CHO (Chinese hamster ovary). Compounds 1 and 2 did not show any antifungal and cytotoxic activity. Compounds 3 and 4 displayed moderate cytotoxic activities, as well as 4 antifungal activity. (C) 2010 Phytochemical Society of Europe. Published by Elsevier B. V. All rights reserved.

Avaliação toxicológica da preparação fitoterápica contendo Piper methysticum Forst, piperaceae (Kava Kava ) sobre o desenvolvimento pré-natal de ratos winstar.

A kava kava (Piper methysticum Forst), pertencente à família Piperaceae, é utilizada para diminuir a ansiedade e medo e tratar distúrbios comportamentais. É um fitoterápico utilizado em vários países, entretanto pouco se sabe, sobre seus efeitos no desenvolvimento embrionário. O presente trabalho avaliou o possível efeito teratogênico da formulação fitoterápica contendo Piper methysticum Forst durante o período de organogênese em ratas Wistar. As ratas foram tratadas com 0mg.kg –1 (controle), 5mg.kg –1 ; 35mg.kg –1 e 50mg.kg –1 da preparação fitoterápica (kava kava®), por via oral, do 6° ao 15° dia de prenhez. Os resultados revelaram ausência de toxicidade sistêmica e reprodutiva nas variáveis avaliadas, fundamentados pela ausência de alterações no desenvolvimento ponderal, consumos de ração e água, na massa relativa dos órgãos, nas reabsorções embrionárias, na massa corporal, na vitalidade, no número de fetos por progenitora e nas alterações macroscópicas externas e esqueléticas dos fetos. Adicionalmente as enzimas alanina aminotrasferase (ALT) e fosfatase alcalina (FA) foram determinadas no soro das ratas tratadas, para avaliar o possível efeito hepatotóxico da preparação fitoterápica. Não houve alteração na atividade das enzimas ALT e FA, bem como alterações histopatológicas do fígado das ratas, não confirmando a hepatotoxicidade. Conclui-se que o fitoterápico kava kava, até 35mg.kg –1, não determina o aparecimento de teratogenicidade.

Effects of the Ottonia martiana Miq. (Piperaceae) extract on dog's ocular surface

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro efficacy of the essential oil of Piper cubeba L. (Piperaceae) against Schistosoma mansoni

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ultrastructure and cytochemistry of the pearl gland in Piper regnellii (Piperaceae)

Pearl glands are scattered throughout the lamina of developing leaves and rarely found on adult leaves of Piper regnellii (Piperaceae). The pearl gland is a bicellular secretory trichome composed of a short broad basal cell and a spatula-like, semiglobular apical cell. Four different stages of the pearl grand were determined during its ontogenesis: origin, pre-secretory, secretory and post-secretory. During the pre-secretory stage, mitochondria, ribosomes, dictyosomes, rough endoplasmic reticulum, and plastids with electron dense inclusions were present in the cytoplasm of the apical cell. During the secretory stage, the most remarkable characteristics of the apical cell are the proliferation of dictyosomes and their vesicles, rough endoplasmic reticulum, and modified plastids. At this stage, electron-dense oil drops occur in the plastids as well as scattered within the cytoplasm, proteins and polysaccharides are seen in the plastids, vesicles, and vacuoles. Only polysaccharides are present in the periplasmic space, wall cavities, and on the surface of the apical cell. The polysaccharides are one of the main components of the mucilagenous exudate that covers the developing leaf structures. The apical cell of the senescing trichomes undergoes a progressive degeneration of its cellular components, the plastids being the first organelles to undergo lysis.

Morfoanatomia dos órgãos vegetativos de Piper hispidum Sw. (Piperaceae)

O presente trabalho teve por objetivo o estudo morfoanatômico dos órgãos vegetativos de Piper hispidum, visando a estabelecer características marcantes para a sua identificação e auxiliar estudos taxonômicos e farmacobotânicos. O material vegetal fresco e fixado foi estudado segundo as técnicas usuais de corte e coloração, incluindo análise em MEV. Piper hispidum é um arbusto com caule cilíndrico, nodoso, verde claro, com folhas alternas, ovadas, de cor verde-escura na face adaxial e verde claro na abaxial. Dentre as características anatômicas importantes para sua identificação destacam-se: parênquima cortical da raiz apresentando grupos de esclereídes. Córtex caulinar com faixas descontínuas de colênquima do tipo angular e tecido vascular organizado em dois círculos descontínuos de feixes colaterais. A folha é dorsiventral, hipoestomática, com estômatos tetracíticos. Hipoderme adaxial descontínua e abaxial frouxa com número variável de camadas; tricomas tectores e glandulares ocorrem nas duas faces. Epiderme uniestratificada e idioblastos oleíferos ocorrem em todos os órgãos.