Vouacapane diterpenoids isolated from Pterodon and their biological activities

Abstract The Pterodon genus comprises two native species in Brazil, known as "sucupira-branca" or "faveira". Their fruits have long been used in Brazilian natural medicine, mainly for the treatment of infections and inflammations. The pharmacological properties of these fruits have often been linked with vouacapane diterpenoids. This review evaluated the scientific research in the period from 1973 to February 2017, aiming to answer how difficult it still is to develop a scientifically supported product based on Pterodon vouacapanes. Therefore, this paper reviews purification, identification, and quantification methods applied to vouacapane diterpenoids from Pterodon, as well as the performance of these phytochemicals in pharmacological tests described in the literature. Data analysis results support conventional notions that suggest vouacapane diterpenoids from Pterodon have anti-inflammatory properties. However, the studies carried out so far still represent partial assessment of the vouacapane activities and further studies need to be completed. Pterodon diterpenoids have also been associated with larvicidal, leishmanicidal, cardiovascular, and antitumor activities, which reinforces the genus' potential as a source of phytomedicines. Some remaining gaps about the reviewed activities were mentioned, while trends and perspectives for future research were proposed.

Box-Behnken experimental design for extraction of artemisinin from Artemisia annua and validation of the assay method

ABSTRACT Obtaining artemisinin and its derivatives is very costly, which limits access to low-income people. Some hydroalcoholic extract of Artemisia annua L., Asteraceae, which has shown comparable antimalarial activity could be an alternative to the purified compound, especially if the cultivars have higher content of artemisinin. The objective of this study was to evaluate the effects of the extraction parameters (ethanol graduation, previous shaking time in an ultrasound bath and drug/solvent ratio) on the yield of artemisinin in the liquid extract obtained by percolation from A. annua and then optimize the extraction efficiency of this compound. The Box-Behnken (33) factorial design was used in association with response surface methodology. The derivatization reaction of artemisinin was used in an analytical method which was submitted to validation, after reaching the specification of the selectivity, linearity, precision, accuracy and robustness. Detection and quantification limits were 1.3 and 4.0 µg/ml, respectively. The largest amount of this compound of interest was obtained without any ultrasound bath, with an ethanol graduation of 95% and a drug/solvent ratio of 2%. Drug/solvent ratio was the factor which most influenced extraction efficiency. The maximum range of artemisinin yield was 1.21%. Information obtained in this study can be used for future approaches to determining and extracting artemisinin from A. annua.

Identification of terpenes and phytosterols in Dipteryx alata (baru) oil seeds obtained through pressing

ABSTRACTThe oil from seeds of Dipteryx alata Vogel, Fabaceae, popularly known as baru, was extracted by hydraulic and continuous screw pressing. A total of eleven chemical constituents obtained by hydraulic pressing, including steroids, mono and sesquiterpenes and tocopherol derivatives were identified by gas chromatography–tandem mass spectrometry (GC–MS). Compounds limonene, β-elemene, γ-elemene, α-caryophyllene, β-caryophyllene, campesterol, stigmasterol, β-sitosterol and cycloartenol are being described for the first time in the baru oil.

Environmental factors affecting the concentration of phenolic compounds in Myrcia tomentosa leaves

Myrcia tomentosa (Aubl.) DC., Myrtaceae, found in Central Brazilian Cerrado and popularly known as "goiaba-brava", belongs to the Myrcia genus, which has several species with medicinal properties such as: hypoglycemic, diuretic, hypotensive, antidiarrheal, antimicrobial and antitumor. The present study aimed to analyzed the environmental influence on concentrations of phenolic metabolites in M. tomentosa leaves. Compounds assayed in the leaves were: total phenols, tannins by protein precipitation, hydrolysable tannins and total flavonoids and mineral nutrients, while soil fertility was also analyzed, all over during one year. The results were submitted to Pearson Correlation Analysis and stepwise Multiple Regression Analysis to investigate the relationship between phenolics and environment data. Analysis of variance and Cluster Analysis allowed indicated a high variability in samples from different sites. The results obtained suggests that content of phenolics from M. tomentosa leaves are influenced by environmental factors, particularly some foliar nutrients (N1, Ca1 and Mn1), soil nutrients (Ca s and Ks) and Rainfall.

Spray drying of Eugenia dysenterica extract: effects of in-process parameters on product quality

A 2³ full factorial design was used to assess the impact of spraying air flow rate (30-50 L/min), drying air inlet temperature (90-150 ºC) and extract feed rate (4-6 g/min) on the quality of Eugenia dysenterica DC., Myrtaceae, spray-dried extracts. Response surface methodology (RSM) was applied to analyze the significance of the effects of process factors on product quality and to obtain fitted equations to predict dry powder properties. Powder yields were satisfactory, ranging from 34.64 to 63.92%. The dried products showed moisture contents and water activities below 5% and 0.5, respectively. The recuperation ratios of total polyphenols, tannins and flavonoids ranged from 88.66 to 99.07%, 70.38 to 81.87% and 74.51 to 98.68%, respectively. Additionally, in some conditions the parameters related to dry product’s flowability and compressibility varied over a range acceptable for pharmaceutical purposes. RSM proved that studied factors significantly affected most of the quality indicators at different levels. The spray drying technology is an attractive and promising alternative for the development of intermediate phytopharmaceutical products of E. dysenterica.

Mechanism involved in the anti-inflammatory effect of Spiranthera odoratissima (Manacá)

Acetic acid-induced writhing, hot-plate, carrageenan-induced pleurisy, formalin-induced pain, croton oil-induced ear edema, vascular permeability tests and phospholipase A2 activity assay were used to study the analgesic and/or anti-inflammatory activity of the hydromethanolic fraction of ethanolic extract from Spiranthera odoratissima A. St.-Hil., Rutaceae, leaves (HMF) and its subfraction (sub-Fr10-28). HMF and sub-Fr10-28 reduced the leukocyte migration on the carrageenan-induced pleurisy test; sub-Fr10-28 reduced the pain reaction time in the second phase of formalin-induced pain, as well as the ear edema and vascular permeability. Both HMF and sub-Fr10-28 inhibited the phospholipase A2 activity. These results suggest that the analgesic effect of this plant could be, in part, due to an anti-inflammatory action produced by the inhibition of phospholipase A2 activity.

Influence of environmental factors on the concentration of phenolic compounds in leaves of Lafoensia pacari

Lafoensia pacari A. St.-Hil., Lythraceae, a plant from the Cerrado known as pacari or dedaleiro, is widely used as an antipyretic, wound healing, anti-inflammatory, antidiarrheal and in the treatment of gastritis and cancer. Notable among the metabolite groups identified in leaves of L. pacari are the polyphenols, such as tannins and flavonoids, related to the pharmacological activities of pacari. Studies on the influence of environmental factors over production of major groups of secondary metabolites in pacari are important because they contribute data for its cultivation and harvest, and establish quantitative parameters of secondary metabolites in the plant drug. The objective of this study was to evaluate the influence of environmental factors on concentrations of phenolic metabolites in the leaves of L. pacari. Compounds quantified in the leaves were: total phenols, tannins by protein precipitation, hydrolysable tannins, total flavonoids, ellagic acid and mineral nutrients, while soil fertility was also analyzed, all over a period of one year. The data were analyzed using multivariate analysis, and the results suggest that metabolite concentrations in the leaves of this plant are influenced by seasonal factors, in particular the temperature and foliar micronutrients (Cu, Fe, Mn, Zn).

Dissolution test of herbal medicines containing Passiflora sp.

The dissolution test is an essential tool to assess the quality of herbal medicines in the solid dosage form for oral use. This work aimed to evaluate the dissolution behavior of three herbal medicines in the form of capsules and tablet containing Passiflora, produced with powder or dried extract. Assay of total flavonoids and dissolution methods were validated and obtained results allowed the quantification of flavonoids with precision, accuracy and selectivity. The percentage of total flavonoids found was 2 percent for capsule A (containing only powder), 0.97 percent for capsule B (containing only dried extract) and 5.5 percent for tablet. Although the content was lower, the release of flavonoids present in the capsule containing dried extract was 12 percent higher over 30 min, with dissolved percentage values of 87 and 75, for the capsules containing extract and powder, respectively. The tablet containing dried extract presented dissolution of 76 percent, despite the higher content of flavonoids, which may be due to pharmacotechnical problems. Obtained data demonstrated the need to implement these tests in the quality control of herbal medicines, confirming the release of the active ingredients that underlie the pharmacological action of these medicines.

Composição química, atividade antimicrobiana do óleo essencial e ocorrência de esteróides nas folhas de Pterodon emarginatus Vogel, Fabaceae / Chemical composition, antimicrobial activity of essential oil and the occurrence of steroids in the leaves of Pterodon emarginatus Vogel, Fabaceae

Pterodon emarginatus Vogel, Fabaceae, conhecida popularmente como sucupira-branca, é uma espécie arbórea que possui interesse como planta medicinal e fonte de madeira. Vários estudos com o óleo e extratos dos seus frutos demonstraram atividades cercaricida, antimicrobiana e antiinflamatória. Tendo em vista a importância de P. emarginatus como planta medicinal o presente trabalho analisou a composição química, a atividade antifúngica e antibacteriana do óleo essencial de suas folhas, isolou e identificou compostos do extrato etanólico bruto das folhas. A análise do óleo essencial das folhas permitiu a identificação de 9 hidrocarbonetos sesquiterpênicos, sendo os majoritários o γ-muuroleno (48,79 por cento) e o biciclogermacreno (22,66 por cento). O óleo essencial apresentou atividade frente a bactérias Gram-positivas, sendo inativo sobre isolados clínicos de Candida. O estudo fitoquímico do extrato etanólico das folhas permitiu o isolamento da mistura dos esteróides estigmasterol e β-sitosterol. Todos os resultados obtidos contribuem para a ampliação de informações sobre essa planta amplamente utilizada pela população.

Pterodon emarginatus Vogel, Fabaceae, known as "sucupira-branca", is a tree specie with interest as medicinal plant and source of wood. Several studies with the oil and fruit's extracts have been showed cercaricid, antimicrobial and antiinflamatory activities. The importance of P. emarginatus conducted the present study to realize evaluated antifungal and antibacterial activity of essential oil of their leaves, and isolated some compounds of ethanolic extract of the leaves. The analysis of the essential oil from leaves allowed the identification of nine sesquiterpenics hydrocarbons, with the majority γ-muurolene (48.79 percent) and biciclogermacrene (22.66 percent). The essential oil showed activity against the Gram-positive bacteria, and inactive on clinical isolates of Candida. The phytochemical study of the ethanol extract of the leaves allowed the isolation of the mixture of stigmasterol and β-sitosterol steroids. All results contribute to the expansion of information about this plant widely used by the population.

Morfoanatomia de Memora nodosa (Silva Manso) Miers, Bignoniaceae / Morpho-anatomy of Memora nodosa (Silva Manso) Miers, Bignoniaceae

Memora nodosa (Silva Manso) Miers, Bignoniaceae, é uma planta do Cerrado utilizada popularmente como cicatrizante de úlceras e feridas externas (folhas e caules). O objetivo deste trabalho foi caracterizar morfologicamente essa espécie e fazer o estudo anatômico de suas folhas. O estudo morfológico foi realizado à vista desarmada e com o auxílio de microscópio estereoscópico. Para o estudo anatômico as folhas foram seccionadas e as lâminas histológicas confeccionadas de acordo com as técnicas convencionais. Memora nodosa é um arbusto com folhas opostas cruzadas, recompostas, bipinadas, imparipinadas, com foliólulos pequenos, lanceolados, verde-escuros e brilhantes. A inflorescência é racemosa com flores infundibuliformes e corola amarelo-ouro; glândulas nectaríferas são observadas no terço superior do cálice; androceu com quatro estames férteis, didínamos e um estaminódio reduzido; gineceu com estilete terminal, ovário súpero, assentado sobre um disco nectarífero. O fruto é do tipo cápsula septífraga achatada, com sementes aladas. Anatomicamente a lâmina do foliólulo é hipoestomática com estômatos predominantemente paracíticos. O parênquima paliçádico é pluriestratificado, ocupando cerca de dois terços do mesofilo dorsiventral. Tricomas glandulares e tectores são observados, em maior número, nas folhas jovens.

Memora nodosa (Silva Manso) Miers, Bignoniaceae, is a Brazilian Savannah plant, popularly used as external wound and ulcer healing (leaves and stems). The goal of this research was to perform the morphological study of M. nodosa species and the anatomical study of its leaves. The morphological study was done to the naked eye and with the aid of a stereoscopic microscope. For the anatomical study leaves were sectioned and histological slides made in accordance with conventional techniques. Memora nodosa is a shrub with decussate leaves recomposed, bipinnate, imparipinnate with small leaflets, lanceolate, dark green and shining. The inflorescence is racemose with infundibuliform flowers and gold yellow corolla; nectar glands are observed in the upper third calyx; androecium with four fertile stamens, didynamus and one staminode; gynoecium with terminal style, superior ovary, seated on a nectary disc. The fruit is flattened septifragal capsule with winged seeds. Anatomically, the blade leaflet is hypostomatic with predominantly paracytic stomata. The palisade parenchyma is pluriseriate, occupying about two thirds of the dorsiventral mesophyll. Glandular and non-glandular trichomes are observed in greater numbers in the young leaves.

Atividade citotóxica e antiangiogênica de Punica granatum L., Punicaceae / Cytotoxic and antiangiogenic activities of Punica granatum L., Punicaceae

Punica granatum L., Punicaceae, amplamente usada no Brasil, foi avaliada quanto ao seu potencial antitumoral in vitroe in vivo. Investigou-se in vitro a citotoxicidade do extrato etanólico do fruto e folha da P. granatumutilizando células K-562 e células do Tumor Ascítico de Ehrlich (TAE), pelos métodos de redução do MTT e exclusão do azul de tripano. Nos estudos in vivoavaliou-se o aumento da sobrevida de animais portadores do TAE e tratados, por via oral, com diferentes doses dos extratos etanólicos da P. granatum(12,5; 25; 50 e 100 mg/kg) por dez dias consecutivos. Além disso, nestes animais analisou-se o potencial de inibição tumoral e a atividade antiangiogênica da P. granatum. Os resultados dos estudos in vitrodemonstraram uma redução na viabilidade das células K-562 e do TAE, concentração-dependente, nos métodos investigados. Os resultados in vivo demonstraram aumento da sobrevida dos animais portadores do TAE tratados, de forma dose-dependente. Em paralelo, observou-se diminuição do número de células tumorais na cavidade peritoneal dos animais portadores e tratados. Além disto, os tratamentos empregados reduziram o padrão de vascularização da parede abdominal. Dessa forma, os dados apresentados revelaram que o extrato de P. granatumpossui atividade antitumoral in vitroe in vivo em paralelo a redução da angiogênese peritoneal.

Punica granatum L., a plant widely used in Brazil, was tested for its antitumor and antiangiogenic activities in vitroand in vivo. In this work, the in vitrocytotoxicity was evaluated using the K-562 cell line and Ehrlich ascites tumour cells, by MTT tetrazolium reduction test and the trypan blue exclusion test. In vivostudies investigated the increase in the survival time of Ehrlich tumour-bearing mice after treatment with different doses of Punica granatumL. ethanol extract (12.5; 25; 50 e and 100 mg/kg), by gavages, for ten consecutive days. In addition, we also investigated the tumour inhibition potential and antiangiogenic activity of this plant. In vitroresults demonstrated a decrease of K-562 and Ehrlich ascites tumour cells viability, with both methods used, in a dependent-manner concentration. In vivoresults showed a significant antitumor activity against Ehrlich ascites tumour growth, increasing survival time. In parallel, we detected a significant inhibition of the tumour growth, along with a decrease in the vascular pattern of the peritoneal wall. Thus, the data presented herein clearly showed that Punica granatum L. has antitumor and antiangiogenic activities.

Caracterização farmacognóstica das folhas e sementes de Albizia lebbeck (L. ) Benth. (Fabaceae) / Pharmacognostic characterization of leaves and seeds of Albizia lebbeck (L. ) Benth. (Fabaceae)

Albizia lebbeck (L.) Benth. é popularmente conhecida no Brasil por coração-de-negro. É uma espécie aclimatada nesse país. As sementes dessa espécie são utilizadas no tratamento de diarréia, disenteria e hemorróidas e as folhas, no tratamento de úlceras e nas conjuntivites alérgicas. Este trabalho foi realizado com a finalidade de determinar as características macroscópicas e microscópicas das folhas e sementes dessa planta, uma vez que estas são as partes usadas popularmente. Algumas características de valor na sua diagnose foram apontadas e ilustradas por fotomicrografias. Reações de fitoquímica e a determinação do conteúdo de água e de cinzas foram realizadas no pó das folhas e sementes de A. lebbeck. Saponinas, alcalóides e cumarinas foram detectados nas folhas e sementes; flavonóides foram também identificados nas folhas. O teor de umidade foi 6,78 por cento e 7,91 por cento, respectivamente, para o pó das folhas e sementes. Os valores de cinzas totais e cinzas insolúveis em ácido foram 9,60 por cento e 0,62 por cento para as folhas e 4,13 por cento e 0,21 por cento para as sementes. Os dados obtidos são importantes para o controle de qualidade dessa planta medicinal. O extrato bruto da planta não apresentou atividade antibacteriana.

Albizia lebbeck (L.) Benth., is popularly known in Brazil as "coração-de-negro". It is a species acclimatized in Brazil. The seeds of this species are used to treat diarrhea, dysentery and hemorrhoids and the leaves are used for the treatment of ulcers and allergic conjunctivitis. The purpose of this work was to determine the macroscopic and microscopic characteristics of the leaves and seeds of this plant, since these are the parts used popularly. Some characters of diagnostical value are pointed out and illustrated by photomicrographs. Phytochemistry reactions and determination of water and ash were done on the powder of A. lebbeck. Saponins, alkaloids, coumarins were detected in the leaves and seeds; flavonoids were also detected in the leaves. The content of water was 6.78 percent and 7.91 percent, respectively, for leaf and seed powder. The values of total ashes and acid insoluble ashes were 9.60 percent and 0.62 percent for leaf, and 4.13 percent and 0.21 percent, for seed powder. The data obtained are important for the quality control of this medicinal plant. The crude extract of this plant did not show antibacterial activity.

Determinação do teor de princípios ativos em matérias-primas vegetais / Determination of active principles in vegetable raw materials

Castanha-da-índia (Aesculus hippocastanum L.), hipérico (Hypericum perforatum L.), guaraná (Paullinia cupana H.B.K.) e ginkgo (Ginkgo biloba L.) fazem parte de formulações de medicamentos fitoterápicos muito comercializados no Brasil. Neste trabalho foi avaliada a qualidade de amostras de pós e extratos destas plantas. Para a determinação do teor de princípios ativos foram aplicados métodos farmacopéicos ou citados na literatura científica e os resultados demonstraram serem adequados para a análise destes produtos farmacêuticos. Verificou-se que 60 por cento das amostras analisadas de guaraná e ginkgo estavam em desacordo com as especificações farmacopéicas ou as declaradas pelos laudos dos fabricantes, em relação ao teor de princípios ativos. Além disso, 100 por cento e 20 por cento, respectivamente, das amostras de hipérico e de castanha-da-índia apresentaram teores abaixo das especificações. Os dados obtidos mostraram ser de grande importância e necessidade a implementação de técnicas quantitativas no controle de qualidade físico-químico de matérias-primas vegetais.

Pharmaceutical preparations containing Aesculus hippocastanum L., Hypericum perforatum L., Paullinia cupana H.B.K. and Ginkgo biloba L. have large market. In this work the quality of the powder or extracts of these plants were evaluated. Pharmacopoeia methods or those cited on cientific literature for quantification of active substances were employed and the data showed them to be adequate for these analyses. The analytical results demonstrated that 60 percent of the samples of P. cupana and G. biloba, and 100 percent and 20 percent, respectively of samples of H. perforatum and A. hippocastanum were not in agreement with the specifications. This analysis showed to be valuable for herbal medicines quality control.