Schistosomicidal Evaluation of Zanthoxylum naranjillo and its Isolated Compounds against Schistosoma mansoni Adult Worms

Chemical investigation of the EtOAc fraction (EF) obtained from the ethanolic extract of Zanthoxylum naranjillo (Rutaceae) leaves (EE) by preparative HPLC resulted in the isolation of protocatechuic acid (1), gallic acid (2), p-hydroxybenzoic acid (3), and 5-O-caffeoylshikimic acid (4). This is the first time that the presence of compounds 1-4 in Z. naranjillo has been reported. Compounds 1-4, the EE, and EF were tested in vitro against Schistosoma mansoni adult worms. The results showed that the S. mansoni daily egg production decreased by 29.8%, 13.5% 28.4%, 17.7%, 16.3%, and 6.4%, respectively. Compounds 1 and 3 were also able to separate adult worm pairs into male and female. This activity may be correlated with the reduction in egg production, since 1 and 3 showed better inhibitory properties compared with 2 and 4.

Cubebin and derivatives as inhibitors of mitochondrial complex I. Proposed interaction with subunit B8

The effects on mitochondrial respiration and complex I NADH oxidase activity of cubebin and derivatives were evaluated. The compounds inhibited the state 3 glutamate/malate-supported respiration of hamster liver mitochondria with IC50 values ranging from 12.16 to 83.96M. NADH oxidase reaction was evaluated in submitochondrial particles. The compounds also inhibited this activity, showing the same order of potency observed for effects on state 3 respiration, as well as a tendency towards a non-competitive type of inhibition (KI values ranging from 0.62 to 16.1M). A potential binding mode of these compounds with complex I subunit B8, assessed by docking calculations, is proposed.

(-)-Hinokinin-loaded poly(d,l-lactide-co-glycolide) microparticles for Chagas disease

The (-)-hinokinin display high activity against Trypanosoma cruzi in vitro and in vivo. (-)-Hinokinin-loaded poly(d,l-lactide-co-glycolide) microparticles were prepared and characterized in order to protect (-)-hinokinin of biological interactions and promote its sustained release for treatment of Chagas disease. The microparticles contain (-)-hinokinin were prepared by the classical method of the emulsion/solvent evaporation. The scanning electron microscopy, light-scattering analyzer were used to study the morphology and particle size, respectively. The encapsulation efficiency was determined, drug release studies were kinetically evaluated, and the trypanocidal effect was evaluated in vivo. (-)-Hinokinin-loaded microparticles obtained showed a mean diameter of 0.862 A mu m with smooth surface and spherical shape. The encapsulation efficiency was 72.46 A +/- 2.92% and developed system maintained drug release with Higuchi kinetics. The preparation method showed to be suitable, since the morphological characteristics, encapsulation efficiency, and in vitro release profile were satisfactory. In vivo assays showed significant reduction of mice parasitaemia after administration of (-)-hinokinin-loaded microparticles. Thus, the developed microparticles seem to be a promising system for sustained release of (-)-hinokinin for treatment of Chagas disease.

Cytotoxic evaluation of essential oil from Zanthoxylum rhoifolium Lam. leaves

Zanthoxylum rhoifolium Lam is a plant popularly used as antimicrobial, for malaria and inflammatory treatment. The essential oil of Z. rhoifolium was extracted and its cytotoxic effects against HeLa (human cervical carcinoma), A-549 (human lung carcinoma), HT-29 (human colon adenocarcinoma), Vero (monkey kidney) cell lines and mice macrophages were evaluated. Some of the terpenes of its essential oil (ß-caryophyllene, alpha-humulene, alpha -pinene, myrcene and linalool) were also tested to verify their possible influence in the oil cytotoxic activity. The results obtained permitted to confirm that the essential oil is cytotoxic against tumoral cells (CD50 = 82.3, 90.7 and 113.6 µg/ml for A-549, HeLa e HT-29 cell lines, respectively), while it did not show cytotoxicity against non-tumoral cells (Vero and mice macrophages). Thus, the essential oil from Z. rhoifolium leaves seems to present a possible therapeuthic role due to its selective cytotoxic activity against tumoral cell lines.

Investigation phytochimique d'une brosse à dents africaine Zanthoxylum zanthoxyloides (Lam.) Zepernick et Timler (Syn. Fagara zanthoxiloides L.) (Rutaceae)

Résumé: Dans le but de rechercher de nouveaux composés naturels à intérêt thérapeutique, les extraits dichlorométhanique et méthanolique de Zanthoxylum zanthoxyloides (Lam.) Zepernick et Timler (Syn. Fagara zanthoxyloides L.) (Rutaceae), une brosse à dents africaine ont été soumis à un criblage chimique et biologique. Un dépistage des activités: antifongiques contre le champignon phytopathogène Cladosporium cucumerinum et la levure commensale responsable de mycoses chez l'homme Candida albicans, antibactérienne contre la bactérie opportuniste Bacillus subtilis, larvicide contre le moustique vecteur de la fièvre jaune Aedes aegypti et molluscicide contre Biomphalaria glabrata, un escargot impliqué dans la transmission de la schistosomiase urinaire a été réalisé. Les propriétés antiradicalaires et inhibitrices de l'acétylcholinestérase de ces extraits ont aussi été dépistées. Sur la base des résultats obtenus lors de ce screening, l'investigation phytochimique de ces extraits a été entreprise. Elle a abouti à l'isolement de 14 composés, actifs pour la majorité contre Cladosporium cucumerinum et Bacillus subtilis, dont la structure a été établie au moyen de méthodes spectroscopiques (UV, MS, IR, 1H- et 13C-NMR). Des méthodes chimiques (hydrolyse, acétylation) ont été requises pour la confirmation de structures. L'extrait dichlorométhanique a fourni un nouveau composé, un dérivé du phényléthane, ainsi que dix composés connus, dont trois dérivés du phénylpropane, un lignane, un alcaloïde de la famille des benzophénanthridines, un triterpène, deux amides phénoliques et deux amides oléfmiques. L'extrait méthanolique a fourni un nouveau composé avec une fonction endoperoxyde, qui avait montré une activité inhibitrice modérée de l'acétylcholinestérase, ainsi que l'hespéridine et un dérivé de la chélérythrine. Par ailleurs, l'analyse LC/UV/APC1-MS de cet extrait a permis de détecter on-une sept produits connus. Parmi ces composés, se trouvent l'acide divanilloylquinique, la chélérythrine et quatre de ses dérivés: norchélérythrin.e, 6-(2-oxybutyl) dihydrochélérythrine, 6-hydroxy-dihydrochélérythrine et avicine, ainsi qu'une amide phénolique, l'amottianamide. La présence de ces dérivés de la chélérythrine a été mise en évidence dans deux autres espèces du même genre lors d'une étude LC/UV/APCI-MS comparative. Les activités fongicides contre Cladosporium cucumerinum et Candida albicans et bactéricides contre Bacillus subtilis et Streptococcus mutans ATCC 25175, mises en évidence sur plaque CCM et par les tests de dilution dans l'agar de ces composés, permettent de justifier l'utilisation de Zanthoxylum zanthoxyloides (Lam.) Zepemick et Timler comme brosse à dents africaine. Les techniques couplées de pointe utilisées dans cette étude ont montré leur apport inestimable dans le domaine de la recherche phytochimique et les applications futures dans le domaine de déréplication d'extraits bruts. Abstract: With the aim of discovering new natural therapeutics, the dichloromethane and methanol extracts of the African toothbrush tree Zanthoxylum zanthoxyloides (Lam.) Zepernick et Timler (Syn. Fagara zanthoxyloides L.) (Rutaceae), were submitted to biological and chemical assays. The former included: the antifimgal activities of the extracts against the phytopathogenic fungus Cladosporium cucumerinum, the commensal yeast which causes human mycoses Candida albicans, the bactericidal activity against the opportunistic bacteria Bacillus subtilis, the larvicidal activity against the yellow fever-transmitting mosquito Aedes aegypti and the molluscicidal effect on the snail Biomphalaria glabrata involved in the transmission of urinary schistosomiasis. The antiradical and acetylcholinesterase-inhibiting properties of these extracts were also investigated. On the basis of these results, a phytochemical investigation of the dichloromethane and methanol extracts of Zanthoxylum zanthoxyloides was undertaken. Their fractionation led to the isolation of 14 compounds, the majority of which were active against Cladosporium cucumerinum and Bacillus subtilis, whose structures were elucidated by spectroscopic techniques (UV, MS, IR, 1H- and 13C-NMR). Chemical methods (hydrolysis, acetylation) were performed to confirm the structures. The dichloromethane extract yielded a new phenylethane derivative, together with ten known compounds: three phenylpropane derivatives, a lignan, a benzophenanthridine alkaloid, a triterpene and four phenolic and olefinic amides. The methanol extract yielded a new compound with an endoperoxide moiety, which showed moderate acetylcholinesterase-inhibiting activity, together with hesperidin and a chelerythrine derivative. Seven more compounds were detected on-line by LC/UV/APCI-MS. Among the compounds detected were divanilloylquinic acid, chelerythrine and four chelerythrine derivatives: norchelerythrine, 6-(2-oxybuty1)-dihydrochelerythrine, 6-hydroxy dihydrochelerythrine and avicine, together with the phenolic amide amottianamide. Most of the chelerythrine derivatives were also found in two other Zanthoxylum species following LC/UV/APCI-MS analysis. The antifungal activities against Cladosporium cucumerinum and Candida albicans and antibacterial activities against Bacillus subtilis and Streptococcus mutans ATCC 25175, may explain the utilization in traditional medicine of the roots of this plant as a toothbrush. The advanced hyphenated techniques used in this study showed their inestimable contribution to the field of phytochemical research and applications in the field of dereplication of crude extracts.

Constituintes químicos de Zanthoxylum ekmanii (URB.) Alain

Chemical investigation of Z. ekmanii resulted in the isolation of skimmianine, dictamnine, tembamide, sesamin, lupeol and beta-sitosterol. The structures were established by spectroscopic analyses. This is the first report on the phytochemical study of the roots and leaves of Z. ekmanii.

Alcalóides e outros metabólitos do caule e frutos de Zanthoxylum tingoassuiba A. St. Hil

Phytochemical investigation of this species, popularly known as tinguaciba and used in traditional medicine to various diseases, resulted in the isolation of 15 substances: 2 alkaloids - norchelerythrine and arnottianamide; 1 lignan - sesamin; 4 terpenoids - citronellyl acetate, lupeol, α-bisabolol and spatulenol; 5 coumarins described for the first time - xanthotoxin, isopimpinelin, O-prenylumbelliferone, imperatorin and aurapten, 1 protoalkaloid - methyl N-methylanthranilate and 2 steroids - stigmasterol and β-sitosterol. The structures of the compounds were elucidated by spectroscopic analyses and compared with literature data.

Estudo fitoquímico das cascas das raízes de Zanthoxylum rigidum Humb. & Bonpl. ex Willd (Rutaceae)

Chemical investigation from root barks of Z. rigidum, resulted in the isolation of lupeol, a mixture of steroids campesterol, sitosterol, stigmasterol, sacarose, hesperidin, N-methylatanine and 6-acetonyldihydrochelerythrine. Their structures were established by spectral data analysis. No previous work has been reported on Z. rigidum species.

Constituintes químicos voláteis das folhas e galhos de Zanthoxylum syncarpum Tull.

The essential oils of the leaves and twigs from Zanthoxylum syncarpum Tull. were examined by GC/MS and GC-FID. Variation in the oil composition relative to the harvesting time was also described. The major components in the leaves oils were limonene (23.1-47.3%) and myrcene (4.8-10.8%). In the oils of twigs, the main components were ar-curcumene (12.8-18.1%), E-β-farnesene (9.1-9.7%) and β-caryophyllene (9.2-9.3%). This paper describes for the first time the composition of the essential oil of the twigs from Z. syncarpum.

Chemical constituents of the wood from Zanthoxylum quinduense Tul. (Rutaceae)

Phytochemical investigation of the wood from Zanthoxylum quinduense Tul. allowed the isolation and identification of norchelerythrine, decarine, 6-acetonyldihydrochelerythrine, syringaresinol, evofilin C, p-hydroxybenzaldehyde, vanillic acid, a mixture of β-sitosterol, stigmasterol and campesterol and a mixture of saturated and unsaturated fatty acids, and their esters derivatives. The structures of the isolated compounds were elucidated by spectroscopic techniques and comparison with literature data and the mixture of sterols and fatty acids were identified by GC/MS. The antifungal activity of the ethanolic extract, fractions and pure compounds against Fusarium oxysporum f. sp. lycopersici was determined by bioautography. Evofilin C and nochelerytrine were the only substances that present antifungal activity.

Alcaloides das cascas das raízes de Zanthoxylum spp

In 1959, Gottlieb and Antonaccio published a study reporting the occurrence of lignan sesamin and triterpene lupeol in Zanthoxylum tingoassuiba. In this work we describe the phytochemical study of the root bark of the Z. tingoassuiba which allowed the identification of the lupeol, sesamin, and alkaloids dihydrochelerythrine, chelerythrine, anorttianamide, cis-N-methyl-canadine, predicentine, 2, 3-methylenedioxy-10,11-dimethoxy-tetrahydro protoberberine. The investigation of hexane and methanol extracts of the root bark of Z. rhoifolium and Z. stelligerum also investigated showed the presence of alkaloids dihydrochelerythrine, anorttianamide, cis-N-methyl-canadine, 7,9-dimethoxy-2,3-methylenedioxybenzophenanthridine and angoline. The occurrence of 2,3-methylenedioxy-10,11-dimethoxy-tetrahydro protoberberine is first described in Z. tingoassuiba and Z. stelligerum. This is also the first report of the presence of hesperidin and neohesperidin in roots of Z. stelligerum.

Allelopathy and phytotoxicity of Zanthoxylum rhoifolium Lam.

In Cerrado, Zanthoxylum rhoifolium Lam. stands out because it interferes with the growth and development of other plants. The preliminary phytochemical analyses showed the presence of polyphenols and flavonoids. The Z. rhoifolium extract inhibited the germination index (5 mg mL(-1)=0.5%; 10mg mL(-1)=37.(5%) and 20mg mL(-1)=85.5%) decreased the germination and the lettuce seedlings growth (roots and the hypocotyls) and reduced the mitotic index than control. The extract of 1000 mu g mL(-1)conc contained (165.20 mg and 45.74 mg, phenols and total flavonoids, respectively) and had higher antioxidant activity (80.52%). We concluded that Z. rhofolium possesses allelopathic potential.

Reduction of parasitism tissue by treatment of mice chronically infected with Trypanosoma cruzi with lignano lactones

The reduction of parasitism tissue upon treatment with two lignano lactones, namely (-)- cubebin (CUB) and (-)-hinokinin (HNK), was evaluated in the chronic phase of Chagas` disease by quantifying the enzyme beta-galactosidase expressed by the CL B5 clone strain of Trypanosoma cruzi. Tissue karyometry was also performed. Treatment with the assessed lignans led to a larger reduction in parasitism tissue in all evaluated organs, compared with benznidazole (BZN). Oral treatment with CUB or HNK was more effective. Karyometry results demonstrated that the infected control animals had increased nuclear area compared with uninfected controls, indicating cellular hypertrophy. Results also revealed that use of CUB or HNK was able to significantly prevent this increase, and a slight decrease in the nuclear area was observed, compared with mice treated with BZN. Taken together, these data demonstrate that CUB and HNK could be considered as potential compounds for the development of new drugs for treatment of Chagas` disease.

Composição florística da vegetação arbórea de um trecho de floresta estacional semidecídua em Viçosa, Minas Gerais, e espécies de maior ocorrência na região

É grande a demanda por estudos ecológicos em florestas que possam embasar trabalhos de recuperação, conservação da biodiversidade e apoio à legislação ambiental. Entre esses estudos, primordialmente está o levantamento da flora. Nesse sentido, este trabalho teve como objetivo determinar a composição florística de um trecho da Reserva da Biologia, em Viçosa, MG, em regeneração natural há 80 anos, bem como realizar uma análise comparativa com outros trabalhos conduzidos na região, fornecendo, assim, informações que poderão embasar estudos sobre recuperação e conservação da biodiversidade dos fragmentos de Floresta Estacional Semidecidual. Os dados florísticos foram obtidos mediante a amostragem fitossociológica em 1 ha, pelo método de parcelas contíguas, em que todos os indivíduos arbóreos com circunferência de tronco >15 cm a 1,30 m do solo foram amostrados. Registraram-se 130 espécies, distribuídas em 94 gêneros, pertencentes a 38 famílias botânicas. A análise desses dados conjuntamente com de outros levantamentos realizados em trechos distintos da Reserva da Biologia, no campus da UFV, denota a marcante influência de variáveis ambientais locais (topografia, declividade, face de exposição solar e disponibilidade hídrica) na distribuição da riqueza florística desse fragmento. Casearia decandra, Guapira opposita, Apuleia leiocarpa, Dalbergia nigra, Jacaranda macratha, Matayba elaeagnoides, Piptadenia gonoacantha, Bathysa nicholsonii, Carpotroche brasiliensis, Luehea grandiflora, Mabea fistulifera, Ocotea odorifera, Sorocea bonplandii e Zanthoxylum rhoifolium foram as espécies de maior ocorrência nos fragmentos estudados na região de Viçosa e adjacências, apresentando potencial para uso na recuperação florestal de áreas degradadas.

Estrutura de espécies arbóreas sob efeito de borda em um fragmento de floresta estacional semidecidual em Pernambuco

Os fragmentos florestais são considerados os únicos redutos detentores de biodiversidade do planeta. Conhecer os processos que decorrem após a fragmentação, a exemplo da estrutura arbórea ocorrente na borda, é de fundamental importância para se proporem medidas conservacionistas. Os objetivos deste trabalho, desenvolvido em uma área de 83,8 ha, localizada no Município de Nazaré da Mata, PE, foram efetuar o levantamento fitossociológico de espécies arbóreas adultas sob efeito de borda e verificar a similaridade florística entre as parcelas. A área amostral foi de 10.000 m², equivalentes à implantação de 10 transectos de 10 x 100 m perpendiculares à borda, distribuídos de forma sistemática. Foram amostrados, etiquetados e identificados todos os indivíduos arbóreos com CAP e" 15 cm. Posteriormente, realizaram-se os cálculos dos parâmetros fitossociológicos e da similaridade florística. Neste estudo, amostraram-se 1.238 indivíduos, pertencentes a 72 táxons, distribuídos em 26 famílias botânicas. As espécies Campomanesia xanthocarpa, Zanthoxylum rhoifolium e Anadenanthera colubrina apresentaram o maior valor de importância, pois, teoricamente, conseguiram explorar melhor o recurso proporcionado pelo hábitat.