Taxifolin stability:In silico prediction and in vitro degradation with HPLC-UV/UPLC-ESI-MS monitoring / 药物分析学报

Taxifolin has a plethora of therapeutic activities and is currently isolated from the stem bark of the tree Larix gmelinni(Dahurian larch).It is a flavonoid of high commercial interest for its use in supplements or in antioxidant-rich functional foods.However,its poor stability and low bioavailability hinder the use of flavonoid in nutritional or pharmaceutical formulations.In this work,taxifolin isolated from the seeds of Mimusops balata,was evaluated by in silico stability prediction studies and in vitro forced degradation studies(acid and alkaline hydrolysis,oxidation,visible/UV radiation,dry/humid heating)monitored by high performance liquid chromatography with ultraviolet detection(HPLC-UV)and ultrahigh perfor-mance liquid chromatography-electrospray ionization-mass spectrometry(UPLC-ESI-MS).The in silico stability prediction studies indicated the most susceptible regions in the molecule to nucleophilic and electrophilic attacks,as well as the sites susceptible to oxidation.The in vitro forced degradation tests were in agreement with the in silico stability prediction,indicating that taxifolin is extremely unstable(class 1)under alkaline hydrolysis.In addition,taxifolin thermal degradation was increased by humidity.On the other hand,with respect to photosensitivity,taxifolin can be classified as class 4(stable).Moreover,the alkaline degradation products were characterized by UPLC-ESI-MS/MS as dimers of taxifolin.These results enabled an understanding of the intrinsic lability of taxifolin,contributing to the development of stability-indicating methods,and of appropriate drug release systems,with the aims of preserving its stability and improving its bioavailability.

In vivo and in vitro anti-inflammatory activity of Litchi chinensis Sonn leaf extract and isolated compound procyanidin A2

Litchi chinensis Sonn seeds and pericarp are used in popular medicine as antioxidant, anticancer, antiviral, analgesic, antipyretic, immunomodulatory agent and others. However, literature on litchi leaves extract and the main compound procyanidin A2 (PA2) activities are scarce. Here, we investigated the anti-inflammatory activity of L. chinensis leaves extract and the main compound PA2 using in vitro and in vivo assays. We used the pleurisy and air-pouch model induced by carrageenan, substance P, histamine, bradykinin or lipopolysaccharide (LPS) in mice. The PA2 effects were also evaluated using carrageenan-induced pleurisy and LPS-induced air-pouch. In addition, LPS-induced NO2- production and cytotoxicity were quantified using peritoneal neutrophils recruited by oyster glycogen previously treated with L. chinensis extract or PA2. Animals orally treated with L. chinensis leaves extract exhibited a reduction on carrageenan-induced paw edema. Furthermore, the extract reduced the leukocyte migration and the protein leakage in the pleurisy model induced by carrageenan, substance P, histamine and bradykinin. The main compound in the leaves extract, PA2 reduced the paw edema and cell migration into the carrageenan-inflamed tissue. The data obtained in the LPS-induced air pouch model show that the extract and PA2 inhibit the in vivo neutrophil migration. In addition, the leaves extract and PA2 did not affect the neutrophil viability and reduces NO2- production. Taken together, the data herein obtained showed the modulatory actions of L. chinensis leaves extract on leukocyte migration to inflamed tissue, suggesting its therapeutic application to acute inflammatory process. In addition, it may be supposed that PA2 could be related with these actions, as it is the main compound of L. chinensis leaves extract

Antiproliferative potential of solidagenone isolated of Solidago chilensis

ABSTRACT Plants are considered among the main sources of biologically active chemicals. The species Solidago chilensis Meyen, Asteraceae, is native to the southern parts of South America, where the aerial parts of the plant are commonly used for the treatment of inflammatory conditions. However, the effects of S. chilensis on human cancer cells remain to be elucidated. In this study, we evaluated the antiproliferative effects of the hydroalcoholic and dichloromethane extracts of S. chilensis, as well as their chemical constituents quercitrin and solidagenone against the five human tumor cell lines in vitro. The dichloromethane extract showed a promisor antiproliferative effects in vitro, especially against glioma cell line. Besides, the hydroalcoholic extract and quercitrin were inactive. The diterpene solidagenone showed highly potent antiproliferative effects against breast (MCF-7), kidney (786-0), and prostate cancer (PC-3) cells (total growth inhibition: TGI < 6.25 µg/ml). Solidagenone meets the theoretical physico-chemical criteria for bioavailability of drugs, according to the "Rule of Five" and, by theorical studies, the observed biological effects were probably related to the interaction of the molecule with nuclear receptors and as an enzymatic inhibitor. This study contributes to chemical study and to the identification of antiproliferative molecules in S. chilensis.

Perfil fitoquímico e atividade gastroprotetora dos frutos de Eugenia mattosii / Phytochemical profile and gastroprotective activity of eugenia mattosii fruits

ABSTRACT BACKGROUND: Extracts obtained from plants and fruits provide a relatively safe and practical alternative for the conventional medicine of gastrointestinal diseases. The specie Eugenia mattosii, popularly known in Brazil as "cerejinha", belongs to Myrtaceae family. Species of this family present pharmacological properties, and can be used in the treatment of gastrointestinal disorders. OBJECTIVE: The aim of this study was to determine the phytochemical profile and evaluate the gastroprotective activity of Eugenia mattosii fruits. METHODS: Phytochemical analysis was carried out by thin layer chromatography and gastroprotective assays were performed using two experimental models: acute ulcer model induced by ethanol/HCl and acute ulcer model induced by non-steroidal anti-inflammatory drug (indomethacin). Total lesion area (mm2) and relative lesion area (%) were determined. RESULTS: The results of the phytochemical analysis indicated that the bark and pulp and seeds of E. mattosii present phenolic compounds, terpenes and/or steroids. In gastric ulcer model induced by ethanol was evidenced significant reduction of damaged areas for doses of 50 and 250 mg/ kg of seeds methanol extract, while in the indomethacin-induced ulcer model, all parts of the fruit presented defense capability of the gastric mucosa by reducing lesions at doses of 50, 125 and 250 mg/kg. CONCLUSION: The results demonstrate that the specie E. mattosii has bioactive compounds that provide gastroprotective activity, presenting possible therapeutic potential.

RESUMO CONTEXTO: Extratos obtidos de plantas e frutos fornecem uma alternativa relativamente segura e prática para os remédios convencionais de doenças gastrointestinais. A espécie Eugenia mattosii, popularmente conhecida no Brasil como "cerejinha", pertence à família Myrtaceae. Espécies desta família apresentam propriedades farmacológicas e podem ser utilizadas no tratamento de distúrbios gastrointestinais. OBJETIVO: O objetivo deste estudo foi determinar o perfil fitoquímico e avaliar a atividade gastroprotetora dos frutos de Eugenia mattosii. MÉTODOS: A análise fitoquímica foi realizada por cromatografia em camada delgada e dois modelos experimentais foram utilizados para avaliação da atividade gastroprotetora em camundongos: modelo de úlcera gástrica induzida por anti-inflamatório não-esteroidal (indometacina) e modelo de úlcera gástrica induzida por etanol/HCl. RESULTADOS: Os resultados da análise fitoquímica indicaram que a casca e polpa e as sementes de E. mattosii apresentam compostos fenólicos, terpenos e/ou esteroides. No modelo de úlcera gástrica induzido pelo etanol, foi evidenciada redução significativa de áreas danificadas para doses de 50 e 250 mg/kg do extrato das sementes, enquanto no modelo de úlcera induzida por indometacina, todas as partes do fruto apresentaram capacidade de defesa da mucosa gástrica ao reduzir as lesões nas doses de 50, 125 e 250 mg/kg. CONCLUSÃO: Os resultados demonstram que a espécie E. mattosii possui compostos bioativos com atividade gastroprotetora, apresentando possível potencial terapêutico.

Bioactive dicaffeoylquinic acid derivatives from the root extract of Calea urticifolia

ABSTRACT Calea urticifolia (Mill.) DC., Asteraceae, is a native plant of the Yucatan Peninsula used in traditional medicine to treat inflammation and pain. The bioassay-guided purification of the ethanol root extract allowed the isolation of the main bioactive metabolites, which were identified as an inseparable mixture of thymol (1) and 3-methyl-4-isopropylphenol (2), together with 3,4-O-dicaffeoylquinic acid methyl ester (3), 3,4-O-dicaffeoyl-epi-quinic acid methyl ester (4), 3,5-O-dicaffeoyl-epi-quinic acid methyl ester (5) and 3,5-O-dicaffeoylquinic acid (6). The results showed that the analgesic activity detected in the root extract of C. urticifolia could be attributed mainly to the mixture of 1 and 2 and to the novel 3,4-O-dicaffeoyl-epi-quinic acid methyl ester (4). Alternatively, the similarity on the antiinflammatory and antioxidant activities of the dicaffeoylquinic acid derivatives 3-5 suggests that the former might be related to their ability as radical scavengers.

Phytochemical profile and gastroprotective potential of Myrcianthes pungens fruits and leaves

BACKGROUND: The last decade has been marked by increasing data regarding gastroinstestinal diseases, specially gastritis and ulcer. In order to prevent or treat these diseases, many studies have demonstrated the potential of medicinal plants. The aim of this study was to evaluate the phytochemical profile and the gastroprotective activity of the methanolic extract of Myrcianthes pungens whole fruit, peel, pulp, seeds, and leaves. METHODS: The methanolic extracts were analyzed by thin layer chromatography (TLC) to detect the presence of phenolic compounds by direct comparison with an authentic sample. To evaluate the gastroprotective activity, two experimental models were used: acute ulcer model induced by ethanol/HCl and acute ulcer model induced by nonsteroidal anti-inflammatory drug (indomethacin). Animals were divided in different groups (n= 6) and pretreated orally with the methanolic extracts of M. pungens at doses of 50, 125, and 250 mg/kg, the positive control (cimetidine 100 mg/kg) and negative control (distilled water).RESULTS: The TLC analysis indicated the presence of the flavonoids quercetin and quercitrin in the leaves, quercetinin the peel, and catechin and epicatechin in the leaves and seeds of M. pungens. The extracts of leaves, peel, and pulp showed significant gastroprotective potential regarding the relative area of the lesion observed only in acute ulcer model induced by ethanol. The extracts of whole fruit, peel, pulp, seeds, and leaves showed significant gastroprotective potential observed in acute ulcer induced by indomethacin model. CONCLUSIONS: The gastroprotective activity can be related with the presence of some phenolic compounds identified in phytochemical analysis.

Screening of wild fruit trees with gastroprotective activity in different experimental models / Triagem de plantas frutíferas silvestres com ação gastroprotetora em modelos in vivo

ABSTRACT BACKGROUND Given the increase of people with gastrointestinal disorders, the search for alternative treatments with fewer side effects is vital, as well as the demand for food or plants that can help protect the stomach. OBJECTIVE The aim of this study was to evaluate the gastroprotective action of the extracts of wild fruit trees of Myrcianthes pungens (guabiju); Inga vera Willd. (ingá-banana) and Marlierea tomentosa Cambess. (guarapuruna) in in vivo pharmacological models. METHODS The different parts of the fruits were separately subjected to a process of extraction by methanol. Two experimental pharmacological models were conducted in mice; the gastric ulcer model induced by non-steroidal anti-inflammatory (indomethacin), and the gastric ulcer model induced by ethanol/HCl, which allowed us to evaluate the gastroprotective activity of the extracts at a dose of 250 mg/kg. Subsequently, the total lesion area (mm2) and relative lesion area (%) were determined. RESULTS The results showed significant gastroprotective activity against the aggressive agents used - ethanol and indomethacin - for all the extracts tested. CONCLUSION It is assumed that the fruits have bioactive compounds such as antioxidant substances that act on the prostaglandin levels, protecting them from the damage caused by ethanol and indomethacin. These results prompt further studies to isolate and identify the active properties.

RESUMO CONTEXTO Devido ao aumento de pessoas com distúrbios gastrointestinais, a busca de tratamentos alternativos com menos efeitos cola­terais é fundamental, assim como a demanda por alimentos ou plantas que possam ajudar a proteger o estômago. OBJETIVO O presente estudo teve como objetivo avaliar a ação gastroprotetora dos extratos plantas frutíferas silvestres Myrcianthes pungens (guabiju); Inga vera Willd. (ingá-banana) e Marlierea tomentosa Cambess. (guarapuruna) em modelos farmacológicos in vivo. MÉTODOS As diferentes partes do fruto foram submetidas se­paradamente a um processo de maceração em solução metanólica a frio. Foram realizados dois modelos experimentais em camundongos, modelo de úlcera gástrica induzida por anti-inflamatório não-esteroidal (indometacina) e modelo de úlcera gástrica induzida por etanol/HCl, que permitiram avaliar a atividade gastroprotetora dos extratos na dose de 250 mg/kg. Posteriormente, foram determinadas a área total de lesão (mm2) e a área relativa lesada (%). RESULTADOS Os resultados apresentaram atividade gastroprotetora significativa para todos os extratos estudados frente aos agentes agressores utilizados, etanol e indometacina. CONCLUSÃO Supõe-se que os frutos apresentam compostos bioativos, como as substancias antioxidantes, que atuam sobre os níveis de prostaglandinas, protegendo dos danos causados pelo etanol e indometacina. Os resultados encorajam futuros estudos para isolamento e identificação dos princípios ativos dos frutos.

Simultaneous determination of epicatechin and procyanidin A2 markers in Litchi chinensis leaves by high-performance liquid chromatography

ABSTRACT The fruits of Litchi chinensis Sonn., Sapindaceae, are renowned for their biological activities. However, their leaves are poorly explored, although they represent an important source of vegetable raw material with biological properties as antioxidant, anti-inflammatory and antinociceptive. An HPLC method was developed and validated for the simultaneous quantification of epicatechin and procyanidin A2 in the leaf hydroethanolic extract of L. chinensis. The markers and other unidentified components were separated on a Luna Phenomenex C18 column (250 mm × 4.6 mm, 5 µm) with mobile phase composed of acetonitrile: water pH 3.0 (with sulfuric acid), in a gradient run; at 1.0 ml min-1, 30 ºC and 278 nm for detection. The method was linear over an epicatechin and procyanidin A2 concentration range of 10–100 µg ml-1. The Limit of Quantification for epicatechin and procyanidin A2 were 1.7 and 2 µg ml-1, respectively. The Relative Standard Deviation (%) values for markers (intra- and inter-day precision studies) were <4.0% and the accuracy was 100 ± 5%. The method was applied to ten samples collected in the state of Santa Catarina (Brazil), which showed 14.8–44.5 and 44.8–69.6 mg g-1 of epicatechin and procyanidin A2, respectively. The proposed method could be a valuable tool for quality assessment of L. chinenis leaves as well as their herbal derivatives.

Development of an oral suspension containing dry extract of Aleurites moluccanus with anti-inflammatory activity

Abstract Aleurites moluccanus L. (Willd.), Euphorbiaceae, is a tree that is native to Indonesia and India. Various parts of this tree are commonly used in traditional medicine to treat pain, fever, inflammation, hepatitis, gastric ulcer and other ailments. An oral suspension containing dried extract of A. moluccanus was developed and in vivo anti-inflammatory activity was evaluated. Extract 100 and 50 mg/ml loaded oral suspensions were prepared using different suspending agents. The formulations were analysed by their appearance, pH, density, redispersion time, rate of settling, rheological behaviour, distribution of particle size and zeta potential. The dose uniformity was determined by measuring the content of total phenolic compounds expressed in swertisin by a validated HPLC method, as well as the dissolution profile. The stability of oral suspensions was analysed in accelerated studies (40 °C for 6 months). The anti-inflammatory activity was analysed using an in vivo paw oedema model. The taste and odour of the suspensions were shown to be characteristic of the extract. Carmellose sodium (CS; 0.5%) and microcrystalline cellulose and carmellose sodium mixture (MCCS; 1%) showed better physical behaviour. The content of total phenolic compounds was 1.6 mg/ml and approximately 100% of the total phenolic compounds dissolved within 10 min. During the stability study, the formulations were approved by their physical–chemical properties and were shown to lose 12–14% of total phenolic compounds at 40 °C after 6 months. Suspensions containing 50 mg/ml of standardised dried extract inhibited around 35 ± 7.6% of paw oedema. Formulations containing CS showed more anti-inflammatory activity. Suspensions containing dry extract of A. moluccanus were successfully obtained and showed physical and physical–chemistry properties that were appropriate and characteristic of this dosage form, suitable for administration in paediatric and elderly populations, making this an alternative to tablets.

Seasonal influence and cytotoxicity of extracts, fractions and major compounds from Allamanda schottii

The aim of this research was to evaluate the fractions obtained from the leaf, stem and roots of Allamanda schottii Pohl, Apocynaceae, responsible for the cytotoxicity, using several cell lines. Cytotoxicity was correlated with the season the part of the plant, and the major compounds were assessed. The ethanol extracts of leaves, stems and roots obtained at different seasons were evaluated in the human erythromyeloblastoid leukemia cell line (K562). Subsequently the ethanol extracts and dichloromethane fractions collected in winter were evaluated in mouse fibroblast cell line (Mus musculus) (L929), cervix adenocarcinoma (HeLa), human pre-B leukemia (Nalm6), as well as K562 cell line. The compounds plumericin, plumieride and ursolic acid isolated from ethanol extracts of the stems were evaluated in the same cell lines, as well as on breast adenocarcinoma cell line (MCF-7), and Mus musculus skin melanoma cell line (B16F10). The chromatographic profiles of the dichloromethane fractions were obtained by high performance liquid chromatography. The results revealed that the season during which A. schottii was collected, and the part of the plant analyzed, influence the cytotoxicity on the K562 cells tested. On the other hand the dichloromethane fractions, mainly from the stems and roots, are responsible for the cytoxicity on the cells tested. These results may be associated with the seasonal variation of plumericin in these parts of the plant. This information is in accordance with the HPLC analysis. The results clearly show the potential for the phytotherapeutic use of this species, and suggest that the cytotoxic activity observed may be due to the presence of plumericin, or to minor compounds not yet identified. The seasonal influence on the production of secondary metabolites was verified.

Drimanes from Drimys brasiliensis with leishmanicidal and antimalarial activity

This paper evaluates CHCl3 and CH3OH extracts of the stem bark, branches and leaves of Drimys brasiliensis and drimane sesquiterpenes isolated from the stem bark against strains of Leishmania amazonensis and Leishmania braziliensis promastigotes and Plasmodium falciparum trophozoites. All of the extracts and compounds were tested in cell lines in comparison with reference standards and cell viability was determined by the XTT method. The CHCl3 and CH3OH extracts from the stem bark and branches yielded promising results against two strains of Leishmania, with 50% inhibitory concentrations (IC50 ) values ranging from 39-100 µg/mL. The CHCl3 extract of the stem bark returned IC50 values of 39 and 40.6 µg/mL for L. amazonensis and L. braziliensis, respectively. The drimanes were relatively effective: 1-β-(p-coumaroyloxy)-polygodial produced IC50 values of 5.55 and 2.52 µM for L. amazonensis and L. braziliensis, respectively, compared with 1-β-(p-methoxycinnamoyl)-polygodial, which produced respective IC50 values of 15.85 and 17.80 µM. The CHCl3 extract demonstrated activity (IC50 of 3.0 µg/mL) against P. falciparum. The IC50 values of 1-β-(p-cumaroyloxyl)-polygodial and 1-β-(p-methoxycinnamoyl)-polygodial were 1.01 and 4.87 µM, respectively, for the trophozoite strain. Therefore, the results suggest that D. brasiliensis is a promising plant from which to obtain new and effective antiparasitic agents.

Development of hydroethanolic extract of Ipomoea pes-caprae using factorial design followed by antinociceptive and anti-inflammatory evaluation

Ipomoea pes-caprae (L.) R. Br., Convolvulaceae, is a medicinal plant that grows abundantly as a pan-tropical stand plant. The 3² (two factors and three levels) factorial design, was applied to determine the best time and drug/solvent proportion to maximize the flavonoid content in the hydroethanolic extract by maceration process. The antinociceptive and anti-inflammatory effects were studied at 5-20 mg/kg, i.p., using the writhing test and carrageenan-induced pleurisy models in mice. The optimized extract was able to inhibit more than 50% of abdominal writhing at 20 mg/kg, with 55.88%±2.4 of maximum inhibition. Indomethacin, used as positive control, inhibited 64.86% at 10 mg/kg. In the pleurisy model, the extract produced dose-dependent inhibition of the first phase of inflammation (4 h) in the pleural cavity induced by injection of carrageenan (1%) in mice. It inhibited 50%±0.82 (p<0.01) of exudation induced by carrageenan, and 60.88%±0.14 (p<0.01) of leukocyte migration to the pleural cavity. In conclusion, the results validate the technological conditions of the maceration process to produce an optimized bioactive herb extract for the development of analgesic and anti-inflammatory phytopharmaceuticals using 70 ºGL ethanol, a plant to solvent ratio of 12.5% (w/v), and ten days of maceration.

Atividade antifúngica de imidas cíclicas contra Cryptococcus / Antifungal activity of cyclic imides Cryptococcus neoformans

A criptococose é uma infecção fúngica causada pela levedura Cryptococcus neoformans que acomete tanto hospedeiros normais quanto imunocomprometidos. Nos últimos anos, o número de cepas resistentes a váriso antifúngicos de uso clínico tem apresentado um vertiginoso crescimento. Neste contexto, é de fundamental importância a pesquisa de novas alternativas terapêuticas, através do estudo com produtos naturais, substâncias isoladas e sintéticas, como as imidas cíclicas. O presente trabalho teve como objetivo investigar a atividade antifúngica das imidas cíclicas frente a cepas de Cruptococcus neofornans. A triagem antifúngica foi realizada utilizando a técnica de difusão em meio sólido. Para a determinação da concentração inibitória mínima, foi empregada a técnica da microdiluição. Foram testadas naftalimidas, succimidas e maleimidas. Dentre estas substâncias, as maleimidas ... De acordo com os resultados , as maleimidas surgem como candidatas a antifúngicos por atuarem contra uma espécie de microorganismo que comumente causa infecções.

Anti-inflammatory, antinociceptive, and antipyretic effects of methanol extract of Cariniana rubra stem bark in animal models

Cariniana rubra Miers (Lecythidaceae), popularly known as "jequitibá-vermelho'', is a large Brazilian tree whose bark is used in infusion and decoction for the treatment of inflammatory conditions. This study aims to assess the anti-inflammatory, antinociceptive, and antipyretic effects of Cariniana rubra methanolic stem bark extract (EM Cr) using experimental animals. Anti-inflammatory activity of EM Cr was tested on carrageenan and dextran-induced rat paw edema, carrageenan-induced pleurisy in rats and acetic acid-increase vascular permeability in mice. Antinociceptive and antipyretic activities were evaluated using acetic acid-induced writhing, formalin and hot-plate tests in mice, as well as brewer's yeast-induced pyrexia in rats. The extract inhibitied carrageenan and dextran-induced edema, reduced exudate volume and leukocyte migration on the carrageenan-induced pleurisy and on the vascular permeability increase induced by acetic acid. The EM Cr inhibited nociception on the acetic acid-induced writhing and in the second phase of formalin test, and decreased rectal temperature. It was, however, inactive against thermal nociception.Phytochemical analysis with EM Cr showed the occurrence of saponins, triterpenes, sterols and phenolic compounds. Phytosterols (β-sitosterol, stigmasterol), pentacyclic triterpenes (α- and β-amyrin as a mixture), arjunolic acid, a phytosterol glycoside (sitosterol 3-O-β-D-glucopyranoside), and triterpenoid saponins (28-β-glucopyranosyl-23-O-acetyl arjunolic acid; 3-O-β-glucopyranosyl arjunolic acid and 28-O-[α-L-Rhamnopyranosyl-(1→2)-β-glucopyranosyl]-23- O-acetyl arjunolic acid) were the main identified compounds. It can be presumed that EM Cr caused their effects by inhibiting the liberation and/or action of different inflammatory mediators. These findings support the traditional use of Cariniana rubra preparations to treat inflammation.

Cariniana rubra Miers (Lecythidaceae), popularmente conhecido como "jequitibá-vermelho'', é uma árvore brasileira de grande porte, cuja casca é utilizada nas formas de infusão e decocção para o tratamento de condições inflamatórias. Os efeitos antiinflamatório, antinociceptivo e antipirético do extrato metanólico da casca do caule de Cariniana rubra (EM Cr) foram avaliados em animais experimentais. A atividade antiinflamatória do EM Cr foi testada nos modelos de edema depata induzido por carragenina e dextrana em ratos, pleurisia induzida por carragenina em ratos e permeabilidade vascular aumentada por ácido acético em ratos. As atividades antinociceptiva e antipirética foram avaliadas utilizando os modelos de nocicepções induzidos por ácido acético e formalina, placa quente em camundongos e de pirexia, pela injeção de levedura de cerveja em ratos. O extrato inibiu o edema induzido porcarragenina e dextrana, reduziu o volume de exsudato e a migração de leucócitos na pleurisia induzida por carragenina eo aumento da permeabilidade vascular induzida por ácidoacético. O EM Cr inibiu a nocicepção nas contorções induzidas por ácido acético e na segunda fase do teste de formalina e diminuiu a temperatura retal. No entanto, foi inefetivo no teste da placa quente. A análise química por via úmida deu resultados positivos para saponinas, triterpenos, esteroides e compostos fenólicos. Fitosteróis e triterpenóides pentacíclicos (β-sitosterol, estigmasterol, α and β-amirinas em mistura e ácido arjunólico) e as saponinas triterpenoidais: 3-O-β-D-glucopiranosideo de sitosterol; ácido arjunólico 28-β-glucopiranosila-23-O-acetila; ácido arjunólico 3-O-β-glucopiranosila e ácido arjunólico 28-O-[α-L-rhamnopiranosil-(1→2)-β-D-glucopiranosila]-23-O-acetila. Pode-se presumir que os efeitos do EM Cr foram causados pela inibição da liberação e/ou ação de diversos mediadores inflamatórios. Estes resultados validam o uso tradicional das preparações caseiras de Cariniana rubra para tratar a inflamação.

Evaluation of the antifungal activity and mode of action of Lafoensia pacari A. St.-Hil., Lythraceae, stem-bark extracts, fractions and ellagic acid / Avaliação da atividade antifúngica e modo de ação dos extratos da entrecasca, frações e ácido elágico de Lafoensia pacari A. St.-Hil., Lythraceae

Stem-bark extracts, fractions and the isolated constituent, ellagic acid of Lafoensia pacari St. Hil. (Lythraceae) were in vitro assayed for antifungal activity against a panel of yeasts, hialohyphomycetes as well as dermatophytes with the microbroth dilution method. The EtOH extract and its fractions and ellagic acid exhibited activity against Candida spp and Saccharomyces cerevisiae with MIC values between 250-1000 µg/mL, but they showed no action against filamentous fungi and dermatophytes (MIC>1000 µg/mL). Active extracts were evaluated in Neurospora crassa hyphal growth inhibition and sorbitol assays and then the effect of ergosterol on the MIC of ellagic acid was studied. The active extracts and its fractions and ellagic acid showed a blotchy zone around the paper disk and induced malformations of the hypha. Besides, MIC of the ellagic acid against the Saccharomyces cerevisiae was raised from 62 to 250 µg/mL in the presence of sorbitol 0.8 M, suggesting that the ellagic acid would probably exert its action on fungal cell wall. These results indicate that ellagic acid might be the main active antifungal compound of Lafoensia pacari and further suggest that the mode of antifungal action of these extracts and ellagic acid could be associated with the inhibition of fungal cell wall.

Os extratos, frações e ácido elágico, isolados da entrecasca de Lafoensia pacari A. St.-Hil., Lythraceae, foram testados in vitro para atividade antifúngica, frente a um painel de leveduras, hialo-hifomicetos e dermatófitos, utilizando o método de microdiluição. O extrato EtOH, frações e ácido elágico exibiram atividade contra Candida spp. e Saccharomyces cerevisiae com valores de CIM entre 250-1000 µg/mL, porιm não mostraram ação contra fungos filamentosos e dermatófitos (CIM>1000 µg/mL). Os extratos ativos foram avaliados nos ensaios de inibição do crescimento das hifas de Neurospora crassa, no teste do sorbitol, e pelo estudo do efeito do ergosterol na CIM do ácido elágico. Os extratos ativos, frações e ácido elágico mostraram zonas manchadas ao redor dos discos de papel e induziram malformações nas hifas. Além disso, a CIM do ácido elágico contra Saccharomyces cerevisiae passou de 62 para 250 µg/mL na presença do sorbitol 0,8 M, sugerindo que o ácido elágico provavelmente poderia exercer ação na parede celular fúngica. Esses resultados indicam que o ácido elágico pode ser o principal composto antifúngico de Lafoensia pacari, sugerindo que o modo de ação antifúngico desses extratos e ácido elágico poderia estar associado à inibição da parede celular fúngica.

Evaluation of the proliferative activity of methanol extracts from six medicinal plants in murine spleen cells

A number of natural compounds have been used as immunomodulatory agents, enabling the function of the immune system to be modified by stimulating or suppressing it. There has been increasing interest in the study of therapeutic action of plant extracts regarding their immunomodulatory activity. The aim of this study was to identify and evaluate the action of extracts of the medicinal plants Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis and Vernonia scorpioides on the development of spleen cells from mice, using the in vitro cellular proliferation assay. The cells, obtained by mechanical rupture of mice spleen (5x10(4) cells/mL), were incubated with methanol extracts (10, 50, 100 and 200 µg/mL) and phytohemagglutinin (PHA, 5 µg/mL). The basal control for proliferation consisted of cells alone, while the positive control consisted of cells and PHA. The cell culture was kept at 37 ºC in 5 percent CO2 for 72 hours, and cell proliferation was revealed by the blue tetrazolium reduction assay (MTT). The results were expressed as percentage of growth and were analyzed using the Kruskal-Wallis and Mann-Whitney tests. The C. brasiliense, I. pes-caprae and M. elaeagnoides extracts showed dose-dependent induction of cell proliferation, with a significant increase in cell proliferation (p<0.03) and percentage growth of 88.2 percent, 73.1 percent and 52.7 percent, respectively, suggesting T lymphocyte stimulation. By contrast, M. robusta, R. imperialis and V. scorpioides extracts showed significance only with a negative percentage of growth, suggesting inhibition of cell proliferation (p<0.04). Further biomonitoring studies will enable the fractions and isolated substances responsible for the immunomodulatory activities to be identified.

Várias substâncias de origem natural têm sido utilizadas como agentes imunomoduladores, permitindo modificar a função do sistema imune e propiciando o estudo de atividades terapêuticas de extratos de plantas. Este trabalho objetivou identificar a atividade imunomodulatória dos extratos de seis plantas medicinais da flora brasileira, Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis e Vernonia scorpioides, sobre a proliferação de células esplênicas de camundongos. As células esplênicas murinas obtidas por ruptura mecânica do baço (5x14³ células/mL) foram incubadas com os extratos metanólicos das plantas (10, 50, 100, 200 µg/mL) e fito-hemaglutinina (PHA, 5 µg/mL). O controle basal de proliferação foi constituído de células apenas e o controle positivo formado por células e PHA. O cultivo celular foi mantido a 37 ºC, 5 por cento de CO2, 72 horas, com quantificação da proliferação celular pelo ensaio de redução do azul de tetrazólio. Os resultados expressos em percentagem de crescimento foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney. Os extratos de C. brasiliense, I. pes-caprae e M. elaeagnoides mostraram indução dose-dependente da proliferação celular (p<0,03), com percentagem de crescimento de, respectivamente, 88,2 por cento, 73,1 por cento e 52,7 por cento, sugerindo estímulo de linfócitos T. Contrariamente, os extratos de M. robusta, R. imperialis e V. scorpioides apresentaram significância apenas com percentagem de crescimento negativa, indicando inibição da proliferação celular (p<0,04). A continuidade no estudo biomonitorado permitirá a identificação das frações e substâncias isoladas responsáveis pelas atividades imunomoduladoras.

Caracterização química e tecnológica das folhas, caules e planta inteira da Ipomoea pes-caprae (L.) R. Br., Convolvulaceae, como matéria-prima farmacêutica / Chemical and technological characterization of Ipomoea pes-caprae (L.) R. Br., Convolvulaceae, leaves, branches and entire plant as a pharmaceutical raw material

Ipomoea pes-caprae (L.) R. Br., (Convolvulaceae), planta que ocorre abundantemente nas dunas do litoral é popularmente conhecida como "salsa-da-praia", e usada na medicina popular devido às suas propriedades analgésica, antiinflamatória e cicatrizante. A partir das partes aéreas frescas foram desenvolvidos protocolos para o seu tratamento prévio e controle de qualidade, com o propósito de caracterizar a planta como matéria-prima farmacêutica. A perda por secagem mostrou que as folhas apresentam menor rendimento de matéria seca em relação aos caules, devido ao seu maior teor de água. A secagem e moagem teve por objetivo a manutenção da perda por dessecação abaixo do valor máximo permitido para drogas vegetais. As folhas e caules secos e moídos foram classificados como pós grossos através da análise granulométrica por tamisação, e caracterizados segundo testes farmacopeicos para drogas vegetais. A determinação de cinzas indicou a presença de material inorgânico, possivelmente areia, associado ao habitat da espécie. O teor de extrativos solúveis em água foi maior nas folhas, seguido dos caules e proporcional na planta inteira, correlacionando-se ao teor de flavonoides totais. A cromatografia em camada delgada das soluções extrativas hidroetanólicas mostrou diferenças apenas com relação à intensidade das manchas, e indicação da presença de isoquercitrina, sendo mais evidente nas folhas.

Ipomoea pes caprae (L.) R. Br., (Convolvulaceae), a pantropical stand plant, popularly known as "salsa-da-praia", is used in folk medicine because of its analgesic, antinflammatory and healing properties. Protocols were developed from the fresh aerial plants to establish their previous treatment and quality control in order to get it characterized as a pharmaceutical raw material. The loss of water on drying in an air oven, showed that leaves represent a lesser dried mass yield than branches, as a result of its higher water content. Drying and milling were aimed at keeping the loss on drying below the limit value accepted for herbs. Dried and milled leaves and branches were classified as thick powder through the sieving method, before they were characterized according to pharmacopoeial tests for herbs. The determination of ashes indicated the presence of inorganic materials, most probably due to sand content which came from the herb habitat. The soluble water extractives were greater from the leaves, followed by the branches and proportional for the entire plant. These results showed to be correlated to the total flavonoid content. The thin layer chromatography for the hydroethanolic solutions only demonstrated differences related to the spots color intensity, and indicated the presence of isoquercitrin, being more evident from the leaves.

Antinociceptive effects of Abarema cochliacarpos (B.A. Gomes) Barneby & J.W.Grimes (Mimosaceae) / Efeito antinociceptivo de Abarema cochliacarpos (B.A. Gomes) Barneby & J.W.Grimes (Mimosaceae)

In this study, we investigated the analgesic activity of crude aqueous and methanol extracts obtained from Abarema cochliacarpos bark in mice, and analyzed its phytochemical profile. All the extracts exhibited analgesic properties against the writhing test in mice, but the aqueous and methanol extracts were more active, and more potent than two known analgesic and anti-inflammatory drugs used as reference. They were also active against the capsaicin-model, but inactive when evaluated in the hot-plate test. Phytochemical studies revealed the presence of saponins, catechins, tannins, phenols and anthraquinones.

No presente trabalho foram avaliados a atividade antinociceptiva e o perfil fitoquímico dos extratos aquosos e metanólico produzidos com a casca do caule de Abarema cochliacarpos, uma espécie de Mata Atlântica com diversas indicações populares. Todos os extratos apresentaram atividade analgésica quando avaliados pelo teste das contorções abdominais induzidas pelo ácido acético via intraperitonial, apresentando respostas superiores às drogas usadas como referência, bem como no modelo da dor induzida por capsaicina. A avaliação fitoquímica demonstrou a presença de saponinas, catequinas, taninos, fenóis e antraquinonas.

Antimicrobial screening of some medicinal plants from Mato Grosso Cerrado / Triagem antimicrobiana de algumas plantas medicinais do Cerrado de Mato Grosso

Os extratos em hexano, diclorometano, acetato de etila e etanol das entrecascas de Bowdichia virgilioides, Calophyllum brasiliense, Cariniana rubra, Lafoensia pacari e Stryphnodendron obovatum, do rizoma de Simaba ferruginea e do látex de Croton urucurana foram triados contra um painel de bactérias e fungos usando o método de microdiluição em caldo. O látex de Croton urucurana foi o material derivado de planta com maior atividade antimicrobiana. Os extratos em acetato de etila e hexano da entrecasca de Calophyllum brasiliense destacaram-se por suas seletivas atividades antibacterianas. Os extratos polares da entrecasca de Lafoensia pacari notabilizaram-se por suas potentes e seletivas atividades contra leveduras e os extratos polares e não-polares de Bowdichia virgilioides por suas atividades antifúngicas contra hialo-hifomicetos e dermatófitos. Este é o primeiro relato mostrando atividades antifúngicas para os extratos de Cariniana rubra e Simaba ferruginea. Esse trabalho demonstrou a atividade antimicrobiana de plantas medicinais do Cerrado de Mato Grosso em ensaios in vitro e indica que elas podem ser potenciais candidatas para o desenvolvimento de novas estratégias no tratamento de infecções bacterianas e fúngicas.

Hexane, dichloromethane, ethyl acetate and ethanol extracts from stem barks of Bowdichia virgilioides, Calophyllum brasiliense, Cariniana rubra, Lafoensia pacari, and Stryphnodendron obovatum and rhizome of Simaba ferruginea and Dragon's blood red sap from Croton urucurana were screened against a panel of bacteria and fungi using the micro-broth dilution method. Dragon's blood from Croton urucurana was the most effective antimicrobial plant material. Ethyl acetate and hexane extracts of Calophyllum brasiliense stem bark deserved distinction by their selective antibacterial activity. Lafoensia pacari stem bark polar extracts distinguished by their potent and selective anti-yeast activity and Bowdichia virgilioides polar and non-polar extracts by their antifungal activity towards hyalohypho-mycetes and dermatophytes. This is the first report showing antifungal activity for polar extracts of Cariniana rubra and Simaba ferruginea. This study has demonstrated antimicrobial activity of Mato Grosso Cerrado ethnomedicinal plants in in vitro assays and has indicated that they can be effective potential candidates for the development of new strategies to treat fungal and bacterial infections.

Controle de qualidade de drogas vegetais a base de Bauhinia forficata Link (Fabaceae) / Quality control of drugs with Bauhinia forficata Link (Fabaceae)

Este trabalho teve como objetivo utilizar métodos de controle de qualidade para algumas drogas vegetais a base de Bauhinia forficata comercializadas nos municípios de Itajaí e Balneário Camboriú, utilizando como marcador químico o flavonóide kaempferitrina. As metodologias empregadas foram cromatografia em camada delgada (CCD) e cromatografia líquida de alta eficiência (CLAE), além da análise microscópica, análise de rótulos e características organolépticas e a presença de material estranho. Todas as análises foram comparadas com os resultados demonstrados pela amostra autêntica, devidamente identificada, de B. forficata (A). Foram analisadas 06 amostras a base de Bauhinia identificadas através de siglas, sendo a B. forficata autêntica a amostra A e seis comerciais denominadas B-G. Todas as amostras comerciais foram reprovadas quanto à presença de material estranho, ficando acima de 2 por cento. Na análise microscópica observou-se que apenas a amostra E e G apresentam-se iguais à B. forficata. A análise cromatográfica revelou a presença de kaempferitrina somente nas amostras B, C, E e G.

The present study aimed to use methods to perform the quality control of some vegetal drugs based on Bauhinia forficata from Itajaí and Balneário Camboriú, using the flavonoid kaempferitrin as chemical marker. The methods used were thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), besides microscopic analysis, label analysis and organoleptic characteristics and strange material presence. All the analyses were compared with the authentic sample of B. forficata (A) results. This study used six commercial samples which were denoted as B-G. In the strange material analyses, all the samples were disapproved because they were above 2 percent. Microscopic analyses revealed that only B and G samples were identical to B. forficata (A). The chromatographic results suggested that only the samples B, C, E and G presented kaempferitrin.