Indole monoterpene alkaloids from Chimarrhis turbinata DC Prodr.: a contribution to the chemotaxonomic studies of the Rubiaceae family

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

Determinação das temperaturas cardeais para a germinação de sementes de Murraya paniculata L. Jack. (Rutaceae)

A Murraya paniculata, é conhecida popularmente como murta-de-cheiro ou falsamurta, pertence à família Rutaceae e é uma espécie nativa da Índia que foi introduzida no Brasil. Essa espécie é utilizada na arborização de ruas e em jardins das cidades graças à copa densa e sua resistência a condições adversas. Pode ser utilizada também para formação de cerca vivas e, sua madeira branca tem alta durabilidade podendo ser utilizada em marcenarias. Na Ásia, esta espécie é considerada medicinal e as folhas e raízes são utilizadas para o tratamento de problemas intestinais, de reumatismo e tosse. Do ponto de vista químico, esta espécie acumula principalmente cumarinas e flavonóides, além de derivados do ácido cinâmico e alcalóides. No presente trabalho propomos determinar as temperaturas cardeais para a germinação de sementes de Murraya paniculata, além disso, analisamos a influência do teor de água na germinação para determinar se as sementes são recalcitrantes ou ortodoxas, e a influencia do potencial hídrico induzido por polietilenoglicol (PEG 6000). O aprofundamento dos estudos dessa espécie se faz importante por conta desta ser hospedeira alternativa do psilídeo Diaphorina citri, transmissor do ―greening‖ dos citros, doença que diminuiu a produção de citros nos últimos anos

Produção de biomassa e teor de cumarina em folhas de guaco (Mikania laevigata Sch. Bip. ex Baker) em diferentes idades da planta

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Exposição de pacientes com síndrome de dor miofascial (SDM) à capsaicina: desenvolvimento de forma farmacêutica de uso tópico com possível ação analgésica

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

Produção de biomassa e teor de cumarina em folhas de guaco (Mikania laevigata Sch. Bip. ex Baker) em diferentes idades da planta

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Exposição de pacientes com síndrome de dor miofascial (SDM) à capsaicina: desenvolvimento de forma farmacêutica de uso tópico com possível ação analgésica

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

Atividade antiofídica do decocto das folhas de jatropha gossypiifolia l. frente o veneno de bothrops jararaca

Antiophidic activity from decoct of Jatropha gossypiifolia L. leaves against Bothrops jararaca venom. Snakebites are a serious worldwide public health problem. In Latin America, about 90 % of accidents are attributed to snakes from Bothrops genus. Currently, the main available treatment is the antivenom serum therapy, which has some disadvantages such as inability to neutralize local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies to treat snakebites is relevant. Jatropha gossypiifolia L., a medicinal plant popularly known in Brazil as “pinhão-roxo”, is very used in folk medicine as antiophidic. So, the aim of this study is to evaluate the antiophidic properties of this species against enzymatic and biological activities from Bothrops jararaca snake venom. The aqueous leaf extract of J. gossypiifolia was prepared by decoction. The inhibition studies were performed in vitro, by pre-incubation of a fixed amount of venom with different amounts of extract from J. gossypiifolia for 60 min at 37 °C, and in vivo, through oral or intraperitoneal treatment of animals, in different doses, 60 min before venom injection. The proteolytic activity upon azocasein was efficiently inhibited, indicating inhibitory action upon metalloproteinases (SVMPs) and/or serine proteases (SVSPs). The extract inhibited the fibrinogenolytic activity, which was also confirmed by zymography, where it was possible to observe that the extract preferentially inhibits fibrinogenolytic enzymes of 26 and 28 kDa. The coagulant activity upon fibrinogen and plasma were significantly inhibited, suggesting an inhibitory action upon thrombin-like enzymes (SVTLEs), as well as upon clotting factor activators toxins. The extract prolonged the activated partial thromboplastin time (aPTT), suggesting an inhibitory action toward not only to SVTLEs, but also against endogenous thrombin. The defibrinogenating activity in vivo was efficiently inhibited by the extract on oral route, confirming the previous results. The local hemorrhagic activity was also significantly inhibited by oral route, indicating an inhibitory action upon SVMPs. The phospholipase activity in vitro was not inhibited. Nevertheless, the edematogenic and myotoxic activities were efficiently inhibited, by oral and intraperitoneal route, which may indicate an inhibitory effect of the extract upon Lys49 phospholipase (PLA2) and/ or SVMPs, or also an anti-inflammatory action against endogenous chemical mediators. Regarding the possible action mechanism, was observed that the extract did not presented proteolytic activity, however, presented protein precipitating action. In addition, the extract showed significant antioxidant activity in different models, which could justify, at least partially, the antiophidic activity presented. The metal chelating action presented by extract could be correlated with SVMPs inhibition, once these enzymes are metal-dependent. The phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins, from which the flavonoids could be pointed as major compounds, based on chromatographic profile obtained by thin layer chromatography (TLC). In conclusion, the results demonstrate that the J. gossypiifolia leaves decoct present potential antiophidic activity, including action upon snakebite local effects, suggesting that this species may be used as a new source of bioactive molecules against bothropic venom.

Obtenção de extratos da Opuntia fícus-indica L. Mill e suas aplicações em formulações cosméticas: avaliação in vivo do sensorial e da eficácia hidratante

Opuntia fícus - indica (L.) Mill is a cactacea presents in the Caatinga ecosystem and shows in its chemical c omposition flavonoids, galacturonic acid and sugars. Different hydroglicolic (EHG001 and EHG002) and hydroethanolic subsequently lyophilized (EHE001 and EHE002) extracts were developed. The EHE002 had his preliminary phytochemical composition investigated by thin layer chromatography (TLC) and we observed the predominance of flavonoids. Different formulations were prepared as emulsions with Sodium Polyacrylate (and) Hydrogenated Polydecene (and) Trideceth - 6 (Rapithix® A60), and Polyacrylamide (and) C13 - 14 I soparaffin (and) Laureth - 7 (Sepigel® 305), and gel with Sodium Polyacrylate (Rapithix® A100). The sensorial evaluation was conducted by check - all - that - apply method. There were no significant differences between the scores assigned to the formulations, howe ver, we noted a preference for those formulated with 1,5% of Rapithix® A100 and 3,0% of Sepigel® 305. These and the formulation with 3% Rapithix® A60 were tested for preliminary and accelerated stability. In accelerated stability study, samples were stored at different temperatures for 90 days. Organoleptic characteristics, the pH values and rheological behavior were assessed. T he emulsions formulated with 3,0% of Sepigel® 305 and 1,5% of Rapithix® A60 w ere stable with pseudoplastic and thixotropic behavior . The moisturizing clinical efficacy of the emulsions containing 3,0% of Sepigel® 305 containing 1 and 3% of EHG001 was performed using the capacitance method (Corneometer®) and transepidermal water lost – TEWL evaluation ( Tewameter®). The results showed t hat the formulation with 3% of EHG001 increased the skin moisturizing against the vehicle and the extractor solvent formulation after five hours. The formulations containing 1 and 3% of EHG001 increased skin barrier effect by reducing transepidermal water loss up to four hours after application.

Avaliação dos efeitos do extrato de Passiflora cincinnata Masters em camundongos: efeito na ansiedade e potencial neuroprotetor

Anxiety disorders and Parkinson’s disease (PD) affect a large portion of the world population. Indeed, therapeutic alternatives available do not contribute to improve most clinical conditions and/or are linked with undesirable side effects. Thus, there is a great demand for the development of new drugs to treatment of these diseases. Passiflora cincinnata Mast. is a native species present in several Brazilian states, popularly known as “maracujá do mato”, “maracujá tubarão” or “maracujá mochila”. Additionally, species of Passiflora genus are traditionally known for their exotic flowers, edible fruits with pronounced flavor and for their sedative, tranquilizer and anxiolytic properties reported by folk medicine. These plants possess important organic compounds such as phenols, cyanogenic glycosides, flavonoids and alkaloids, which are responsible for the anxiolytic, antioxidant, anti-inflammatory, antihyperglycemic, among others activities when tested in mammals. Despite this fact, only a few studies have been conducted to investigate the possible in vivo biological effects of Passiflora cincinnata Mast extracts. Thereby, in this study we evaluated the effects of the alcoholic extract of this plant in anxiety and PD animal model. Mice acutely or chronically administered with ethanolic extract of P. cincinnata do not showed any anxiogenic- or anxyolitic-like effect in elevated plus maze (EPM). In order to reproduce PD symptom’s in mice, we administered repeated injections of reserpine which progressively induced motor impairments such as increase in catalepsy, oral movements, and reduction of the average speed of the animals in the open field, as well as depleted dopamine prodution in SNpc cells. Furthermore, this treatment resulted in the loss of aversive memory recall in mice when undergoing PMDAT. Yet, passiflora group also show this amnesic profile. However, animals treated concomitantly with the alcoholic extract of Passiflora cincinnata Mast. showed higher latency for the onset of motor impairment evaluated by catalepsy. Thus, our results shows that the alcoholic extract of the plant P. cincinnata was able to delay the onset of the catalepsy induced by reserpine administration, plus reverted the depletion of dopamine production in SNpc cells.

Avaliação anti-inflamatória e antipeçonhenta das espécies Hancornia speciosa e Mimosa tenuiflora em modelos experimentais de inflamação e envenenamento induzidos por Bothrops jararaca e Tityus serrulatus

Accidents caused by venomous animals represents a significant and serious public health problem in certain regions of Brazil, as well as in other parts of the world by the frequency with which they occur and the mortality they cause. The use of plant extracts as an antidote for poisoning cases is an ancient practice used in many communities that have no access to antivenom. Medicinal plants represent an important source of obtaining bioactive compounds able to assist directly in the treatment of poisoning or indirectly supplementing serum therapy currently used. The aim of this study was to evaluate the effect of extracts, fractions and isolated compounds from M. tenuiflora and H. speciosa in the inflammatory process induced by carrageenan and the venom of B. jararaca and T. serrulatus. The results showed that both M. tenuiflora and H. speciosa were capable of inhibiting cell migration and cytokines levels in peritonitis induced by carrageenin and venom of T. serrulatus. In poisoning by B. jararaca model, mice treated with the plants in studies decreased the leukocyte influx into the peritoneal cavity. Finally the M. tenuiflora and H. speciosa had antiphlogistic activity, reducing edema formation and exerted inhibitory action of leukocyte migration in local inflammation induced by the venom of B. jararaca. Through of Thin Layer Chromatography (TLC) analysis was possible identified the presence of flavonoids ,saponins and/or terpenes in aqueous extract of M. tenuiflora. By High Performance Liquid Chromatography analysis, it was possible to identify the presence of rutin and chlorogenic acid in aqueous extract of H. speciosa. We conclude that the administration of extracts, fractions and isolated compounds of H. speciosa and M. tenuiflora resulted in inhibition of the inflammatory process in different experimental models. This study demonstrates for the first time the effect of M. tenuiflora and H. speciosa in inhibition of the inflammation caused by B. jararaca and T. serrulatus venom.

Estudo fitoquímico de Waltheria ferruginea

The Waltheria genus belonging to the Sterculiaceae family, it is reported as a prolific source of flavonoids and quinolone alkaloids, substances of great interest due to several associated biological activities. This work describes a novel phytochemical study from Waltheria ferruginea, aiming to contribute to the chemical knowledge of this specie and the isolation of substances with biological potential. For the phytochemical study were used chromatography techniques on silica gel and molecular exclusion in Sephadex LH-20.The structural elucidation of the isolated compounds was performed through spectrometric techniques 1H and 13C NMR, including uni and bidimensional pulse sequences, and comparison with data from literature. Five substances were isolated, namely: the flavonids kaempferol-3-O-β-(6''-cumaroil)-glucopyranoside (F1) and kaempferol -3 -O- β - glucopyranoside (F2), both analyzes with pharmacological properties, the flavonol quercetin-3-O-β-glucopyranoside (F3 ) pure and in the epimeric mixture α (F3') and (F3), the terpenegeranyl - geranyl (G1) and the 12-hydroxi-octadecanoic acid, all no previous reported in the literature.

Avaliação dos efeitos tóxicos in vitro e in vivo do extrato hidroetanólico dos frutos de Genipa americana L. (Rubiaceae) em camundongos Swiss

The therapeutic use of medicinal plants has contributed since antiquity in a beneficial way for health. However, many species lacks of scientific evidence which provide basis for their use in therapeutic practice. In this context is the Genipa americana L. species (Rubiaceae), popularly known as jenipapo and used to treat syfilis, ulcer and hemorrhagic disturbs. It's also used against bruising, as tonic and as aphrodisiac. Due this species lacks toxicological studies, the aim of this study was to evaluate the toxicity in vivo (acute and sub-chronic toxicity) and in vitro (cytotoxicity) of the hydroethanolic extract from G. americana fruits. The hydroethanolic extract of G. americana fruits was prepared by maceration. A preliminary phytochemical analysis was performed to assess the presence of secondary metabolites in the extract. The cytotoxicity study of the extract (0.1, 1.0, 10, 100 and 1000 mg / 100 ul) were performed against normal cells (3T3) and tumor (786-0, HepG2 and B16), analyzed by the MTT assay. To evaluate the acute (single dose of 2000 mg / Kg) and subchronic (100, 500 and 1000 mg / kg for 30 days) toxicity Swiss mice of both sexes were used. At the end of the experiment, blood samples and organs were collected for analysis. Data between groups were compared by t test or ANOVA with Dunnett's post-test with 5% significance level. The phytochemical study of the extracts mainly indicated the presence of iridoids. Results for cytotoxicity tests showed up to 70% inhibition of B16 cell line at a dose of 1000 mg / 100 ul, and up to 29% inhibition of 786-0 at a dose of 10 ug / 100 ul. The extract did not cause death in 3T3 and HepG2 cells. During the in vivo assays, there were no animal deaths. Analysis of blood samples revealed that the animals submitted to the evaluation of acute toxicity had changes in AST and ALT, and that the animals evaluated for subchronic toxicity showed changes in the relative wet weight of the kidney and plasma urea concentration. No differences were observed between groups on histopathological evaluation of the collected organs. Despite the changes found in the in vivo toxicity tests, using the criteria described by the OECD Guidelines, it is suggested that the hydroethanolic extract of the fruits of the G. americana is classified as low toxicity. The cytotoxicity of the extract suggests that they have potential against melanoma cell lines (B16).

Avaliação dos efeitos tóxicos in vitro e in vivo do extrato hidroetanólico dos frutos de Genipa americana L. (Rubiaceae) em camundongos Swiss

The therapeutic use of medicinal plants has contributed since antiquity in a beneficial way for health. However, many species lacks of scientific evidence which provide basis for their use in therapeutic practice. In this context is the Genipa americana L. species (Rubiaceae), popularly known as jenipapo and used to treat syfilis, ulcer and hemorrhagic disturbs. It's also used against bruising, as tonic and as aphrodisiac. Due this species lacks toxicological studies, the aim of this study was to evaluate the toxicity in vivo (acute and sub-chronic toxicity) and in vitro (cytotoxicity) of the hydroethanolic extract from G. americana fruits. The hydroethanolic extract of G. americana fruits was prepared by maceration. A preliminary phytochemical analysis was performed to assess the presence of secondary metabolites in the extract. The cytotoxicity study of the extract (0.1, 1.0, 10, 100 and 1000 mg / 100 ul) were performed against normal cells (3T3) and tumor (786-0, HepG2 and B16), analyzed by the MTT assay. To evaluate the acute (single dose of 2000 mg / Kg) and subchronic (100, 500 and 1000 mg / kg for 30 days) toxicity Swiss mice of both sexes were used. At the end of the experiment, blood samples and organs were collected for analysis. Data between groups were compared by t test or ANOVA with Dunnett's post-test with 5% significance level. The phytochemical study of the extracts mainly indicated the presence of iridoids. Results for cytotoxicity tests showed up to 70% inhibition of B16 cell line at a dose of 1000 mg / 100 ul, and up to 29% inhibition of 786-0 at a dose of 10 ug / 100 ul. The extract did not cause death in 3T3 and HepG2 cells. During the in vivo assays, there were no animal deaths. Analysis of blood samples revealed that the animals submitted to the evaluation of acute toxicity had changes in AST and ALT, and that the animals evaluated for subchronic toxicity showed changes in the relative wet weight of the kidney and plasma urea concentration. No differences were observed between groups on histopathological evaluation of the collected organs. Despite the changes found in the in vivo toxicity tests, using the criteria described by the OECD Guidelines, it is suggested that the hydroethanolic extract of the fruits of the G. americana is classified as low toxicity. The cytotoxicity of the extract suggests that they have potential against melanoma cell lines (B16).

Prospeção de compostos bioativos nas macroalgas Himanthalia elongata, Laminaria ochroleuca e Undaria pinnatifida

O presente trabalho teve como principal objetivo caracterizar os metabolitos secundários, compostos lipofílicos e compostos mais polares, de três macroalgas existentes na costa portuguesa, nomeadamente da H. elongata, L. ochroleuca e U. pinnatifida, de forma a contribuir para a sua valorização. Foram realizadas extrações Soxhlet com diclorometano para extrair os compostos lipofílicos, enquanto as frações mais polares foram obtidas por extrações convencionais sólido-líquido, usando diferentes misturas de solventes (acetona:H2O e metanol:H2O:AcOH). Os extratos foram analisados por GC-MS e HPLC-MS. Os extratos polares foram ainda avaliados quanto à sua atividade antioxidante e quanto ao teor de fenóis totais (método de Folin-Ciocâlteau) e florotaninos (método DMBA). A fração lipofílica das três macroalgas estudadas é composta principalmente por ácidos gordos, álcoois alifáticos de cadeia longa e esteróis. O ácido hexadecanóico mostrou ser o composto maioritário das três espécies de algas, seguido dos ácidos octadeca-9-enóico e tetradecanóico. O fucosterol foi o esterol mais abundante encontrado para a H. elongata, enquanto que na L. ochroleuca e na U. pinnatifida foi o 24-metilenocolesterol. Os extratos polares obtidos a partir das duas metodologias de extração apresentaram rendimentos de extração elevados, tendo os extratos acetona:H2O apresentado rendimentos de extração superiores (88.73-92.33 %). Estes extratos mostraram ainda teores de fenóis e florotaninos totais mais elevados, com valores entre 524.03-635.69 g EAG/kg de peso seco e 1.48-1.55 g EFG/kg de peso seco, respetivamente. Os extratos de acetona:H2O no ensaio DPPH apresentaram valores de IC50 entre 6.57-7.64 μg/mL. Estes valores, apesar de serem inferiores ao IC50 do ácido ascórbico, são superiores ao determinado para o antioxidante sintético butil-hidroxitolueno (BHT). O extrato da acetona:H2O da L. ochroleuca foi o que apresentou melhor atividade antioxidante, com um IC50 de 6.57 ± 0.71 μg/mL. A análise por HPLC-DAD-MSn não permitiu até ao momento detetar compostos fenólicos nos extratos obtidos. Estes resultados são uma contribuição relevante para a valorização destas espécies de macroalgas como fonte de fitoquímicos valiosos.

The Addition of Allyltrimethylsilane to Cyclic N-Acyliminium Ions Derived from (S)-(+)-Mandelic Acid and Cyclohexyl-Based Chiral Auxiliaries

A adição de aliltrimetilsilano, promovida por TiCl4, a íons N-aciliminios cíclicos de 5- e 6-membros derivados do ácido (S)-(+)-mandélico, (1R,2S)-trans-2-fenil-1-cicloexanol e (1R,2S,5R)-8-fenilmentol ocorreu com baixas a moderadas razões diastereoisoméricas (1:1-6:1) e forneceu as respectivas amidas e carbamatos em bons rendimentos. A melhor diastereosseleção facial foi observada com o uso de (1R,2S,5R)-8-fenilmentol como auxiliar quiral. As amidas e carbamatos 2-substituídos foram convertidos nos alcalóides (S)- e (R)-propil pirrolidina e coniina com eficiente recuperação dos auxiliares quirais.