Resumo Humiria balsamifera é utilizada na medicina tradicional como antihelmintico, expectorante, para tratar hepatite, diarreia, hemorroidas, na cura de feridas crônicas, e no alívio da dor de dente. Esta espécie ocorre na Restinga de Jurubatiba, no estado do Rio de Janeiro-Brasil, uma região rica que oferece uma variedade de fontes promissoras de produtos bioativos. O presente estudo foca na avaliação química e farmacológica da espécie H. balsamifera. As frações de folhas em n-hexano, diclorometano e acetato de etila apresentaram maior potencial inibitório na produção de NO. Friedelina (1), quercetina (2) e quercetina-3-O-α-arabinopiranosídeo (3) foram isolados e caracterizados; o último está sendo descrito pela primeira vez para H. balsamifera. A quercetina (2) apresentou a melhor atividade inibitória na produção de NO e moderada inibição da produção de TNF-α. Estes resultados contribuem para o conhecimento da espécie Humiria balsamifera como potencial fonte de substâncias anti-inflamatórias. E, ainda, a identificação dos terpenos ß-amirona, betulina, citronelol, eremofileno, dihidroactinolideo e borneol, e o isolamento da quercetina-3-α-O-arabinopiranosideo estão sendo descritos pela primeira vez para esta espécie.

Abstract Humiria balsamifera is used in traditional medicine as anthelmintic, expectorant, to treat hepatitis, diarrhea, hemorrhoids; to cure chronic wounds; and to alleviate toothaches. This species occurs in Jurubatiba shoal, Rio de Janeiro state-Brazil, a rich region which offers a variety of promising bioactive product sources. The present study focuses on the chemical and pharmacological evaluation of H. balsamifera. The n-hexane, dichloromethane and ethyl acetate leaf fractions exhibited higher inhibitory potential on NO production. Friedelin (1), quercetin (2) and quercetin-3-α-O-arabinopyranoside (3) were isolated and characterized; the latter is described for the first time for H. balsamifera. Quercetin (2) showed the best inhibitory activity on NO production and moderate inhibition of TNF-α production. These results contribute to the knowledge of Humiria balsamifera as a source of anti-inflammatory compounds. Furthermore, the identification of the terpenes ß-amyrone, betulin, citronellol, eremophillene, dihydroactinolide and borneol, and the isolation of quercetin-3-α-O-arabinopyranoside are being reported for the first time for this species.

Resumo Ocotea notata (Lauraceae) é popularmente conhecida como canela-branca. As espécies do gênero Ocotea têm vários usos medicinais, especialmente no tratamento de dores no peito, reumatismo e feridas. O presente estudo teve como objetivo analisar a composição química da fração n-hexano de O. notata, além de suas atividades antimicobacteriana e imunomoduladora. A fraçao n-hexano foi analisada por GC-MS e fracionada fornecendo 15 subfrações (SF1-15), onde SF5 foi identificado, por GC-MS e RMN, como o sesquiterpeno espatulenol. A fração n-hexano foi a mais potente na inibição da produção de óxido nítrico (NO) e do fator de necrose tumoral-alfa (TNF-α) em macrófagos estimulados por LPS (IC50 8,3 ± 0,9 e 5,9 ±1,0 μg/mL, respectivamente). SF4, urna subfração majoritária, que apresenta um análogo do espatulenol como constituinte, também inibiu a produção de NO e TNF-α. O espatulenol modulou apenas a produção de NO (IC50 45,6 ±1,4 μg/mL). A fração n-hexano, SF4 e espatulenol apresentaram atividade antimicobacteriana contra as cepas de Mycobacterium bovis BCG, M. tuberculosis (Mtb) H37Rv e M299. O espatulenol inibiu o crescimento de Mtb H37Rv com MIC50 36,9 ± 1,5 μg/mL (167,5 ± 6,8 μM) e Mtb M299 com MIC50 42,1 ± 0,5 μg/mL (191,0 ± 2,2 μM). Este é o primeiro relato que descreve o isolamento do espatulenol das folhas de O. notata e sua atividade antimicobacteriana.

Abstract Ocotea notata (Lauraceae) is popularly known as white-cinnamon. Ocotea species have several medicinal uses, especially for treating chest pain, rheumatism and wounds. The present study aimed to analyze the chemical composition of O. notata n-hexane fraction, in addition to its anti-mycobacterial and immunomodulatory activities. The n-hexane fraction was analyzed by GC-MS and was chromatographed to afford 15 subfractions (SF1-15), where SF5 was identified, by GC-MS and NMR, as the sesquiterpene spathulenol. The n-hexane fraction was the most potent in inhibiting nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α) production on LPS-stimulated macrophages (IC50 8.3 ± 0.9 and 5.9 ±1.0 μg/mL, respectively). SF4, a major subfraction, that presents a spathulenol analogous as a constituent, also inhibited NO and TNF-α production. Spathulenol only modulated NO production (IC50 45.6 ± 1.4 μg/mL). The n-hexane fraction, SF4, and spathulenol revealed antimycobacterial activity against Mycobacterium bovis BCG, M. tuberculosis H37Rv, and M299 strains. Spathulenol inhibited the growth of Mtb H37Rv with MIC50 36.9 ± 1.5 μg/mL (167.5 ± 6.8 μM), and Mtb M299 with MIC5042.1 ± 0.5 μg/mL (191.0 ± 2.2 μM). This is the first report describing the isolation of spathulenol from O. notata leaves and its anti-mycobacterial activity.

Resumo A tuberculose (TB) ainda representa um problema de Saúde Pública em várias regiões do mundo. O tratamento existente é longo e apresenta diversos efeitos adversos. Neste contexto, é relevante a necessidade de identificar novas substancias anti-TB e complementares ao controle do processo inflamatório exacerbado em quadros severos da doença pulmonar. O objetivo deste trabalho foi avaliar, in vitro, extratos e frações de espécies vegetais coletadas na Restinga de Jurubatiba, quanto a sua atividade antimicobacteriana, assim como verificar a ação imunomoduladora e citotóxica em macrófagos. As frações em diclorometano de Kielmeyera membranacea e Eremanthus crotonoides apresentaram os menores CIM50 contra Mycobacterium bovis BCG (0,95 ± 1,08 e 2,17 ± 1,11 μg/mL; respectivamente) e M. tuberculosis H37Rv (4,38 ± 1,19 e 15,28 ± 1,21 μg/mL; respectivamente). Essas também foram capazes de inibir a produção de NO e TNF-α em macrófagos estimulados por LPS, sem serem tóxicas para as células. Através de análise por cromatografia em fase gasosa acoplada ao espectrômetro de massas foi possível sugerir a presença de ácidos graxos e terpenos nas frações mais promissoras, substancias estas descritas por apresentarem atividade antimicobacteriana. Estes resultados permitiram a identificação de Kielmeyera membranacea e Eremanthus crotonoides como as espécies mais promissoras desse estudo, tendo em vista a busca de novos fármacos anti-TB com dupla atividade.

Abstract Tuberculosis (TB) still constitutes a threat to public health in various regions of the world. The existing treatment is long and has many side effects. The need to identify new anti-TB compounds and also adjuvants to control exacerbated inflammation in severe TB cases is relevant. Therefore, the objective of this study was to evaluate the anti-mycobacterial activity of extracts and fractions in vitro from plant species collected in the Restinga of Jurubatiba, in Rio de Janeiro state, Brazil. In addition, to verify their immunomodulatory action and cytotoxicity on macrophages. The dichloromethane fraction of Kielmeyera membranacea and Eremanthus crotonoides showed the lowest MIC50 against Mycobacterium bovis BCG (0.95 ± 1.08 and 2.17 ± 1.11 μg/mL, respectively) and M. tuberculosis H37Rv (4.38 ± 1.19 and 15.28 ± 1.21 μg/mL, respectively). They were also able to inhibit the NO and TNF-α production in LPS-stimulated macrophages, without being toxic to cells. Using gas chromatography analysis coupled with mass spectrometer it was possible to suggest the presence of fatty acids and terpenes in the most promising fractions. Those compounds have been described for their anti-mycobacterial activity. These results have enabled identifying Kielmeyera membranacea and Eremanthus crotonoides as the most promising studied species in searching for new anti-TB compounds with dual activity.

Resumo As restingas são ambientes com características extremas, que fazem parte do bioma da Mata Atlântica. Estes ambientes apresentam características peculiares, como solo arenoso e altas salinidade e exposição solar, o que torna as espécies presentes interessantes para a ciência. A espécie Stachytarpheta schottiana é endêmica do Brasil, presente em restingas e, assim como outras espécies do gênero, apresenta uso na medicina popular. Neste trabalho são descritos pela primeira vez 28 metabólitos secundários da espécie S. schottiana, como iridoides, lignanas, flavonoides e feniletanoides, através do uso de metodologias de desreplicação e molecular networking por LC-HRMS/MS. Destas substâncias, muitas sequer foram descritas para o gênero, como astragalina, taxifolina, lamiídeo e toda a classe de lignanas. Ainda neste trabalho, a Cromatografia Contracorrente de Alta Velocidade (High Speed Countercurrent Chromatography - HSCCC) e a Ressonância Magnética Nuclear (RMN) foram utilizadas para corrigir a identificação equivocada de duas substâncias, bem como para a criação de seeds que seriam posteriormente utilizadas nos molecular networkings. Algumas das substâncias conhecidas sugeridas neste trabalho apresentam atividades biológicas já descritas na literatura que coincidem com o uso popular descrito para a espécie e também para o gênero, como o efeito hepatoprotetor do verbascosídeo.

Abstract Restingas are extreme environments present in the Atlantic Rainforest biome. These ecosystems show peculiar characteristics, like sandy soil, high salinity, and high solar exposure, which brings scientific interest to their native species. Stachytarpheta schottiana is an endemic Brazilian species found in Jurubatiba Restinga, and just like other species of the genus Stachytarpheta, is used in folk medicine. In this paper, we describe, for the first time, 28 secondary metabolites from S. schottiana polar extract, among them iridoids, flavonoids, lignans and phenylethanoids, with the use of LC-HRMS/MS dereplication and molecular networking methodologies. Many of these compounds have not been described yet for the genus Stachytarpheta, like astragalin, taxifolin, lamiide and the lignans secondary metabolites class. Also, in this paper, High Speed Counter-Current Chromatography (HSCCC) isolation and Nuclear Magnetic Resonance (NMR) of two compounds were used to correct a misidentification in the dereplication procedure and to create seeds for molecular networking. Some of the suggested known compounds found in this work have had their biological activities described in the literature, such as the hepatoprotective activity of verbascoside, that matches those already related for the genus Stachytarpheta and for the folk use of Stachytarpheta schottiana itself.

Cancer is one of the most prevalent diseases worldwide and the natural products could be a source of bioactive compounds. Passiflora mucronata (PM) belongs to a very known vegetal genus, although, there are no studies about cytotoxic activity or isolated compounds. Different extracts from PM were obtained by liquid-liquid partition (P), Soxhlet (Sox) and supercritical fluid (SFE1-5) extraction techniques, being compared concerning their yields, chemical profile and cytotoxicity. The Sox extracts showed the highest yields (6.03%: hexane; 2.51%: dichloromethane) followed by SFE (from 4.34 to 1.63%) and partitions (1.06 and 2.26%). The hexane partition (HP) showed the best cytotoxic activity against K562 cell line (IC50 = 18.72 µg.mL-1). From HP, the following compounds were identified and analysed its cytotoxic activities: β-amyrin (IC50 = 3.92 µg.mL-1), β-sitosterol (IC50 = 3.37 µg.mL-1), stigmasterol (IC50 = 3.31 µg.mL-1) and oleanolic acid. Stigmasterol induced about 75% of K562 total apoptosis. The compounds were tested against MA-104 cell line and the selective index (SI) attributed (SI >10 for all compounds). This indicates good selectivity to K562 cell line at the expense of MA-104. This is the first time, identifying those compounds to PM .

ABSTRACT In general, Passiflora species have been reported for their folk medicinal use as sedative and anti-inflammatory. However, P. caerulea has already been reported to treat pulmonary diseases. Severe pulmonary tuberculosis, generally caused by Mycobacterium tuberculosis strains resistant to multiple drugs, can lead to deleterious inflammation and high mortality, encouraging new approaches in drug discovery. Thus, the aim of this work was to evaluate the Passiflora mucronata Lam., Passifloraceae, potential for tuberculosis treatment. Specifically, related to antimycobacterial activity and anti-inflammatory related effects (based on inhibition of nitric oxide, tumor necrosis factor-alpha production and antioxidant potential), as well as the chemical profile of P. mucronata. High performance liquid chromatography coupled with diode-array ultraviolet and mass spectrometer analyses of crude hydroalcoholic extract and ethyl acetate fraction showed the presence of flavonoids. Ethyl acetate fraction showed to be as antioxidant as Ginkgo biloba standard extract with EC50 of 14.61 ± 1.25 µg/ml. One major flavonoid isolated from ethyl acetate fraction was characterized as isoorientin. The hexane fraction and its main isolated compound, the triterpene β-amyrin, exhibited significant growth inhibitory activity against Mycobacterium bovis BCG (MIC50 1.61 ± 1.43 and 3.93 ± 1.05 µg/ml, respectively). In addition, Passiflora mucronata samples, specially hexane and dichloromethane fractions, as well as pure β-amyrin, showed a dose-related inhibition of lipopolysaccharide (LPS)-induced nitric oxide production. In conclusion, Passiflora mucronata presented relevant biological potential and should be considered for further studies using in vivo pulmonary tuberculosis model.