Contribuição ao estudo fitoquímico e biológico de Byrsonima duckeana W. R. Anderson (Malpighiaceae)

Orientador : Prof. Dr. Obdulio Gomes MiguelCo-orientadora : Profª. Drª. Marilis Dallarmi MiguelTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa: Curitiba, 27/11/2014Inclui referênciasÁrea de concentração: Insumos, medicamentos e correlatosResumo: Espécies vegetais são utilizadas ao longo da história da humanidade como recurso para cura e alívio dos mais diversos males. O conhecimento empírico a respeito de diferentes espécies vegetais se tornou uma ferramenta importante na descoberta de substâncias que possam fazer parte do arsenal terapêutico. Baseados nessa premissa o presente trabalho tem como objetivo verificar as informações acerca de atividades biológicas e traçar perfil fitoquímico da espécie vegetal amazônica – Byrsonima duckeana W. R. Anderson. O perfil fitoquímico delineou metabólitos secundários de interesse biológico, tais como a presença de flavonóides, esteróides, terpenos e taninos. O perfil cromatográfico constatou a presença de ácido gálico e as técnicas de elucidação estrutural permitiram que se constatasse a presença do terpeno Friedelina. Na busca por atividades biológicas e farmacológicas, a espécie apresentou atividades de grande interesse, com grande potencial antioxidante e baixa toxicidade, dada a ausência de citotoxicidade no modelo Artemia salina e no ensaio in vivo de toxicidade aguda. Os ensaios farmacológicos demonstraram atividade antiinflamatória e analgésica, em especial para a fração acetato de etila, corroborando com estudos de outras espécies do gênero. Foi encontrada ainda atividade citotóxica no modelo de cultura celular, em células HT29 e atividade hemolítica. Dessa forma, Byrsonima duckeana mostrou ser uma espécie interessante químicamente e farmacologicamente. Palavras-chave: Byrsonima duckeana W.R. Anderson, Farmacologia, Antioxidante, Amazônia, Citotoxicidade.Abstract: Plant species are used throughout the history of mankind as a resource for healing and relief from various ailments . Empirical knowledge about different plant species has become an important tool in the discovery of substances which may form part of the therapeutic arsenal. Based on this premise this study aims to determine information about the biological activities and tracing phytochemical profile of Amazonian plant species - duckeana Byrsonima WR Anderson. The phytochemical profile outlined secondary metabolites of biological interest , such as the presence of flavonoids , steroids , terpenoids and tannins . The chromatographic profile showed the presence of gallic acid and structural elucidation techniques have allowed it appeared the presence of terpene friedelin . In the search for biological and pharmacological activities , the species was of great interest activities with great antioxidant potential and low toxicity , given the absence of cytotoxicity in the model brine shrimp assay and in vivo acute toxicity . Pharmacological studies demonstrated anti-inflammatory and analgesic activity , especially for the ethyl acetate fraction , corroborating studies of other species of the genus . Cytotoxic activity was also found in the cell culture model , in HT29 cells and low hemolytic activity . Thus , Byrsonima duckeana proved to be an interesting specie chemically and pharmacologically. Keywords : Byrsonima duckeana W.R Anderson, Pharmacology, Antioxidant, Amazon, Cytotoxicity

ESTUDO DA INFLUÊNCIA DE JUSTICIA ACUMINATISSIMA (MIQ.) BREMEK. SOBRE A AGREGAÇÃO PLAQUETÁRIA E COAGULAÇÃO SANGUÍNEA

Justicia acuminatissima (Miq.) Bremek (Acanthaceae) é uma espécie vegetal amplamente utilizada pela população do estado do Amazonas devido às suas propriedades medicinais antiinflamatórias. É conhecida popularmente como “sara-tudo” e suas folhas são utilizadas em decoctos, infusões e macerações. Baseado em relatos de seu uso, o presente trabalho avaliou a interferência da preparação extrativa das folhas obtida por maceração aquosa sobre agregação plaquetária e coagulação sanguínea. Nenhuma influência significativa estatisticamente foi observada sobre as duas cascatas, não sendo possível calcular o IC 50

Efeito analgésico e anti-inflamatório do extrato aquoso das folhas de trevo-roxo (Scutellaria agrestis A. St.-Hil. Ex Benth. - Lamiaceae) em roedores

The Scutellaria agrestis is used by Amazonas riverine communities, especially for otitis externa topical treatment, by using the crude extract obtained by maceration. This study aimed to investigate the preliminary phytochemical profile, the safety/toxicity and the analgesic, anti-inflammatory and antiedematogenic activities of the aqueous extract of the S. agrestis leaves. Eighty individuals were collected at the Nilton Lins University medicinal garden, Manaus, Brazil. The phytochemical profile was obtained through a plant drug survey for cyanogenic heterosides, terpenes, alkaloids and phenolic compounds. The extract safety was evaluated by acute toxicity test. Analgesic and anti-inflammatory activities were accessed using formalin test in mice and the antiedematogenic activity, using paw edema test in mice. We detected phenolic (hydrolysable tannins, coumarins and several classes of flavonoids) and terpenoid (free steroids, saponins) metabolites. We could not establish LD50because no animals died during the acute toxicity test, probably because of the absence of cyanogenic glycosides on the composition of the extract. However, we found that the extract is slightly toxic as animal spasms were observed in the first hour of the test. The extract showed significant analgesic and anti-inflammatory activity on the formalin test (30, 100 and 300 mg/kg p.o.), and the highest dose result was equivalent to the standard drug (Fentanyl). However, no significant antiedematogenic effect was observed during the paw edema test. The results obtained in this study provide preliminary scientific basis about the safety and analgesic/anti-inflammatory actions of the aqueous extract of S. agrestis, which indicates that this species is a promising option for further in vitro and in vivo pharmacological studies

CYTOTOXICITY OF BYRSONIMA DUCKEANA W. R. ANDERSON (MALPIGHIACEAE) ON COLON CANCER CELLS

Objective: The objective of this study was the cytotoxicity evaluation of the leaves extract and fractions of Byrsonima duckeana W. R. Anderson. Methods: The toxicity on Artemia salina, haemolytic potential and cytotoxicity activities against two different cell lines a U937 cell line human, and HT29 tumor colon cell line of the leaves extract and fractions of Byrsonima duckeana W. R. Anderson were evaluated. Results: There was no IC50 the Artemia salina assay. The chloroform and ethyl acetate fractions showed haemolytic activity. The results were satisfactory at the most polar fractions, with citotoxicity to the HT29 cell line. Conclusion: The study provides preliminary evidence of cytotoxicity of the most polar fractions at the HT29 cell line, which can indicate a potencial source to offer substances with cytotoxic activity

PHYTOCHEMICAL SCREENING AND PRELIMINARY ASSESSMENT OF ANTIBACTERIAL ACTIVITY OF LEAVES AND STEM BARK EXTRACTS OF Dasyphyllum tomentosum (Spreng.) Cabrera

Dasyphyllum tomentosum (Spreng) Cabrera é conhecida popularmente por açucará, agulheira, espinho-de-agulha e goiapá, pertence à família Asteraceae cujos representantes destacam-se principalmente pelas propriedades terapêuticas. O objetivo deste estudo foi uma investigação qualitativa do perfil químico da planta  a fim de se identificar as principais classes de constituintes presentes nas folhas e casca do caule, além da avaliação preliminar da atividade antibacteriana, tendo em vista a inexistência de estudos na espécie. Para a análise fitoquímica o extrato hidro alcoólico 20% e extrato aquoso foram submetidos reações clássicas de caracterização dos grupamentos químicos, e para avaliação da atividade antibacteriana, o extrato etanólico bruto foi submetido ao ensaio de determinação da Concentração Inibitória Mínima.  A análise fitoquímica apontou a presença de triterpenos e/ou esteroides, flavonoides, alcaloides, heterosídeos antociânicos e taninos. Os extratos etanólicos das folhas e casca do caule de Dasyphyllum tomentosum mostraram ausência de atividade inibitória frente aos microorganismos testados

Phytochemical constituents and preliminary toxicity evaluation of leaves from Rourea induta Planch. (Connaraceae)

A maioria das plantas ativas é tóxica em doses elevadas, portanto, é importante a investigação da toxicidade preliminar dos extratos das plantas. A espécie Rourea induta é uma droga potencial que não apresenta estudo fitoquímico ou biológico descrito na literatura. Assim, um estudo fitoquímico e análises toxicológicas foram realizados com o extrato etanólico obtido das folhas de Rourea induta Planch., Connaraceae. Foram obtidos um hidrocarboneto de cadeia longa, n-tetracosano, e quatro flavonóides, quercetina e três derivados glicosilados, quercetina-3-O-α-arabinofuranosideo, quercetina-3-O-β-xilosideo e quercetina-3-O-β-galactosideo. Esta é a primeira vez que estes compostos são isolados nesta espécie. As estruturas foram elucidadas por espectroscopia de RMN-13C, RMN-¹H, ultravioleta e infravermelho. A avaliação da toxicidade dos extratos foi analisada pelo método da Artemia salina e atividade hemolítica. Nenhuma das amostras testadas apresentou um potencial tóxico pelos métodos analisados.Most active plants are toxic at high doses and it is therefore important to investigate the preliminary toxicity of plant extracts. The Rourea induta species is a potential drug with no phytochemical or biological studies registered in the literature. Thus, a phytochemical study and a toxicity analysis of the ethanolic extract obtained from the leaves of Rourea induta Planch., Connaraceae, was run. A long chain hydrocarbon, n-tetracosane, and four flavonoids were identified: quercetin, and three glycosylated derivates, quercetin-3-O-α-arabinofuranoside, quercetin-3-O-β-xyloside and quercetin-3-O-β-galactoside. This is the first time these have been isolated in this species. The structures were elucidated by 13C NMR, ¹H NMR, UV and IR spectroscopy. The toxicity evaluation of extracts was performed by the brine shrimp method and determination of hemolytic activity. The samples demonstrated no toxic potential by the analyzed methods

Analgesic, Anti-Inflammatory, and Antioxidant Activities of Byrsonima duckeana

Background. Byrsonima is a promising neotropical genus, rich in flavonoids and triterpenes, with several proven pharmacological properties. Nevertheless, Byrsonima duckeana W. R. Anderson is an Amazonian species almost not studied. Objective. To assess the antioxidant, anti-inflammatory, and analgesic activities of Byrsonima duckeana leaves. Materials and Methods. We analyzed an ethanol extract and its fractions for polyphenol content and UHPLC-MS/MS, phosphomolybdenum, DPPH, TBARS antioxidant tests, formalin-induced pain, carrageenan-induced peritonitis, acetic acid-induced abdominal writhings, and hot plate assays. Results. All the samples showed high polyphenol content and antioxidant capacity in the phosphomolybdenum, DPPH, and TBARS tests. We identified ethyl gallate, quinic acid, gallic acid, catechin, epicatechin, quercetrin, and quercetin in the samples. B. duckeana was able to reduce leukocyte migration in the carrageenan-induced peritonitis by 43% and the licking time in the formalin test by 57%. In the acetic acid-induced writhing test, the chloroform (FCL) and ethyl acetate (FEA) fractions were the most active samples. FEA was selected for the hot plate test, where all the dosages tested (5, 50, and 200 mg·kg−1) showed significant analgesic activity. Conclusion. B. duckeana has interesting analgesic and antioxidant activities, due to its high phenolic content, especially phenolic acids

O ESTUDO DE PLANTAS MEDICINAIS E A CORRETA IDENTIFICAÇÃO BOTÂNICA.O ESTUDO DE PLANTAS MEDICINAIS E A CORRETA IDENTIFICAÇÃO BOTÂNICA.

Considerando a importância da correta identificação de plantas com interesse medicinal e problemas relacionados à dificuldade em encontrar especialistas que ajudem na identificação taxonômica, tornou-se necessário abordar o assunto, destacando-se os aspectos sobre coleta, identificação e incorporação do material vegetal no herbário. Apresenta-se aqui uma revisão que reforça a importância da identificação correta das espécies para os trabalhos com produtos naturais, fornecendo algumas ferramentas que venham auxiliar alunos de graduação e de pós graduação

Phytochemical constituents and preliminary toxicity evaluation of leaves from Rourea induta Planch. (Connaraceae)

Most active plants are toxic at high doses and it is therefore important to investigate the preliminary toxicity of plant extracts. The Rourea induta species is a potential drug with no phytochemical or biological studies registered in the literature. Thus, a phytochemical study and a toxicity analysis of the ethanolic extract obtained from the leaves of Rourea induta Planch., Connaraceae, was run. A long chain hydrocarbon, n-tetracosane, and four flavonoids were identified: quercetin, and three glycosylated derivates, quercetin-3-O-α-arabinofuranoside, quercetin-3-O-β-xyloside and quercetin-3-O-β-galactoside. This is the first time these have been isolated in this species. The structures were elucidated by 13C NMR, ¹H NMR, UV and IR spectroscopy. The toxicity evaluation of extracts was performed by the brine shrimp method and determination of hemolytic activity. The samples demonstrated no toxic potential by the analyzed methods

Coumarin isolation and comparative study of biological activities of Pterocaulon alopecuroides DC and Pterocaulon lorentzii Malme

5-(2,3-Dihidroxy-3-methylbuthyloxy)-6,7-methylenedioxycoumarin was isolated from the chloroform extract of the two Asteraceae species Pterocaulon alopecuroides DC. and Pterocaulon lorentzii Malme. The structure was elucidated through IR and 1H and 13C  NMR analyses. The extracts and the isolated compound did not exhibit toxic activity, as determined through the brine shrimp lethality method, and did not interfere with the integrity of erythrocytes, as demonstrated through a hemolytic assay. The antioxidant activities were investigated through three methods. In the phosphomolybdenum test, the ethyl acetate fraction of P. alopecuroides exhibited an antioxidant activity of 137.7% compared with rutin (positive control), and the ethyl acetate fraction of P. lorentzii exhibited an antioxidant activity of 101.7% compared with vitamin C (positive control). The two ethyl acetate fraction also exhibited excellent activity through the DPPH assay: P. alopecuroides and P. lorentzii exhibited IC50 values of 10.74 μg/ml and 7.63 μg/ml, respectively. In the TBARS bioassay, the crude extracts showed the more significant results: IA% 0.419 ± 0.0517 for P. alopecuroides and IA% 0.213 ± 0.0094 for P. lorentzii