Thesis (Ph. D.)--University of Missouri--St. Louis, 2007. / Includes bibliographical references. Also available via the Internet: https://apps.umsl.edu/webapps/weboffice/ETD/query.cfm?id=r2641 (viewed May 19, 2009).

Title from title page of PDF (University of Missouri--St. Louis, viewed March 22, 2010). Includes bibliographical references.

Univ., Diss.--Freiburg/Breisgau, 1996.

Thesis (Ph. D.)--Universität Bern. / Includes bibliographical references.

A espécie Garcinia brasiliensis Mart. (sinonímia: Rheedia brasiliensis Planch e Triana) é uma planta também conhecida como “bacuri”. É uma espécie muito utilizada por suas propriedades farmacológicas. O presente estudo tem a finalidade de caracterizar quimicamente e avaliar a atividade biológica dos extratos etanólicos e aquosos das folhas, cascas e sementes da G. brasiliensis. O teor de compostos fenólicos totais, flavonoides e atividade antioxidante foram avaliados. O extrato que apresentou maior teor de compostos fenólicos foi o extrato aquoso da folha, com 343,98 ± 4,8 mg AG/g. O teores de flavonoides mais expressivos encontrados foram no extrato etanólico da semente e no extrato aquoso da folha, com valores respectivamente de 47,10 ± 0,2 e 45,62 ± 0,7 mg quercetina/g de extrato. Quanto à atividade antioxidante, o que apresentou maior atividade foi o extrato etanólico da semente, com 86,57 % de sequestro de radical DPPH. A atividade antimicrobiana foi avaliada pelo teste de microdiluição em placa para bactérias e fungos. Considerando a CIM50, o extrato que se mostrou mais ativo foi o extrato aquoso da folha, com uma CIM50 de 1 μg/mL, atividade bem próxima ao do padrão cloranfenicol, que possui CIM50 de 0,975 μg/mL contra a bactéria S. aureus. Quanto à atividade antifúngica, constatou-se que nenhum extrato apresentou atividade contra a C. albicans na faixa de concentração testada e com exceção do extrato etanólico da folha, os outros extratos foram considerados bioativos contra a C. krusei. A caracterização química por cromatografia líquida de alta eficiência foi realizada no extrato aquoso da folha e detectou a presença da fukugetina-7’’-O-β-D-glicosídeo (fukugesídeo) e da fukugetina. Com este mesmo extrato realizou-se testes de citotoxicidade em fibroblastos humanos e linhagens tumorais, não apresentando toxicidade para as culturas celulares testadas. Foram preparadas formulações contendo este extrato aquoso de G. brasiliensis e com elas foram realizados testes de permeação e retenção cutânea em “Célula de Franz”. Por ser um produto natural oriundo de uma fonte renovável como as folhas, este extrato é um potencial candidato ao desenvolvimento de novos fármacos para tratar lesões da pele causados pelos microrganismos testados. / The species Garcinia brasiliensis Mart. (synonym: Rheedia brasiliensis Planch and Triana) is a plant also known as "bacuri”. It is a specie commonly used for its pharmacological property. The present study aims to chemically characterize and evaluate the biological activity of ethanol and aqueous extracts of leaves, bark and seeds of G. brasiliensis. The contents of total phenolics, flavonoids and antioxidant activity were evalueted. The leaf aqueous extract showed a high content of phenolic compounds with 343.98 ± 4.8 mg GA / g. The most significant amounts of flavonoids were found in the seed’s ethanolic extract and leaf’s aqueous extract, respectively, with values of 47.10 ± 0.2 and 45.62 ± 0.7 mg of quercetin / g of extract. For antioxidant activity, the most active was the seed’s ethanolic extract, with 86.57 % of scavenging activity. The antimicrobial activity was evaluated by microdilution plate test for bacteria and fungi. Considering the MIC50, the leaf’s aqueous extract was the most active, with a MIC50 of 1 mg / mL, activity very close to chloramphenicol as standard, which showed a MIC50 of 0.975 mg / mL against the bacteria S. aureus. For antifungal activity, no extract showed activity against C. albicans in the range of tested concentrations. Excluding the leaf’s ethanolic extract, all other extracts were considered bioactive against C. krusei. The chemical characterization by high performance liquid chromatography was performed on the leaf’s aqueous extract and it was detected the presence of fukugetin-7''-O-β-D- glucoside (fukugeside) and fukugetin. With this same extract it was carried out cytotoxicity tests in human fibroblasts and tumor cell lines, showing no toxicity of the cell’s cultures tested. Formulations containing G. brasiliensis aqueous extract were prepared and skin permeation and retention tests on "Franz cell" were performed. For being a natural product derived from a renewable source like leaves, this extract is a potential candidate of new drugs development for skin lesions treatment caused by microorganisms tested. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal vi activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections.

Through a continuing search for anticancer compounds as part of an International Cooperative Biodiversity Grant program, the extracts of two plants were selected for study on the basis of their cytotoxic activity. These extracts were further fractionated to yield four compounds. The structures of these compounds were elucidated with mass spectrometry and 1-D and 2-D NMR spectroscopy. The ethyl acetate extract of the twigs of Garcinia macrophylla from Suriname was weakly cytotoxic in the A2780 human ovarian cancer cell bioassay. The known benzophenone guttiferone A and a new guttiferone analog, named guttiferone G, were isolated from the extract and found to be responsible for the bioactivity. A known triterpene, friedelin, was also isolated from the extract and found to be inactive. The structure of guttiferone A was determined by comparison of its NMR data to those found in the literature. The structure of guttiferone G was determined by comparison to guttiferone A and through careful examination of both 1D and 2D NMR data. An extract of Bridelia tulasneana from Madagascar yielded one compound. It was identified as the known lignan deoxypodophyllotoxin and was responsible for the bioactivity. It was identified by a comparison of its spectral data to those found in the literature and those of an authentic sample. / Master of Science

A espécie Garcinia brasiliensis, conhecida como "bacupari", é nativa da Amazônia e cultivada em todo o território brasileiro. Na medicina popular, suas folhas são utilizadas para tratar tumores, inflamações das vias urinárias e artrite, bem como para aliviar dores. Este trabalho foi conduzido para avaliar a atividade analgésica e antiinflamatória do extrato etanólico foliar (GbEE) e da 7-epiclusianona, uma benzofenona natural poliprenilada isolada de suas folhas, em modelos animais in vivo. A ação deste benzofenona também foi investigada em um ensaio antiinflamatório ex vivo, enfocando o “burst” respiratório de neutrófilos e as vias bioquímicas envolvidas. Foi constatada atividade atividade antiinflamatória tanto para GbEE v.o. ( 30, 100 3 300 mg /kg) como para 7-epiclusianona v.o.(5, 10 e 15mg/Kg) por reduzir o edema de pata induzido por carragenina e inibir o recrutamento de leucócitos na cavidade peritoneal. Além disso, através do modelo de indução de inflamação crônica foi observado uma redução estatisticamente significativa de formação do tecido granulomatoso. Por outro lado, efeito antinociceptivo periférico foi atribuído ao GbEE sugerindo ser mais eficaz contra dores inflamatórias, enquanto que para a benzofenona foi constatada tanto atividade antinociceptiva periférica como central, através de diferentes mecanismos. O composto 7-epiclusianona (10 a 100µg) também foi capaz de reduzir e até mesmo suprimir a liberação de ânion superóxido por fagócitos inflamatórios através de um mecanismo controlado pela fosforilação da proteína tirosina e pela estimulação direta da proteína quinase C, sugerindo novas abordagens terapêuticas para o tratamento de processos inflamatórios e desenvolvimento de novos fármacos. Os resultados mostraram atividades antiinflamatória e analgésica das folhas de G. brasiliensis e da benzofenona isolada, confirmando o uso tradicional dessa espécie para tratamento de dores e inflamações. / The species Garcinia brasiliensis, known as “bacupari”, is native to the Amazon and cultivated throughout Brazil. Their leaves are used in folk medicine to treat tumors, inflammation of the urinary tract and arthritis as well as to relief pain. This study was conducted to evaluate the anti-inflammatory and analgesic effects of the ethanolic extract of leaves (GbEE) and the 7-epiclusianone, a natural polyisoprenilated benzophenone isolated from G. brasiliensis, in several animal models. The action of the benzophenone was also investigated ex vivo throughout an anti-inflammatory assay, focusing neutrophil respiratory burst and the biochemical pathway. Antiinflammatory activity was found for both GbEE (30, 100 and 300 mg /kg) p.o. and for 7-epiclusianone (5, 10 and 15mg/kg) p.o. to reduce the paw edema induced by carrageenan and to inhibit leukocyte recruitment into the peritoneal cavity. In addition, through the induction model of chronic inflammation, was observed a statistically significant reduction of granulomatous tissue formation. Moreover, peripheral antinociceptive effect was attributed to GbEE suggesting that this extract could be more effective against inflammatory pain, whereas for benzophenone was verified both peripheral and central antinociceptive activity through different mechanisms.The compound 7-epiclusianone (10 to 100 μg) was also able to reduce and even suppress the release of superoxide anion by inflammatory phagocytes through a mechanism controlled by protein tyrosine phosphorylation and by direct stimulation of protein kinase C, suggesting new therapeutic approaches for the treatment of inflammatory processes and development of new drugs. So, it was demonstrated the anti-inflammatory and antinociceptive activities of the leaves of G. brasiliensis and the benzophenone isolated which supports previous claims of the traditional use of this species against inflammation and pain. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

As plantas são utilizadas no tratamento e na cura de enfermidades desde a antiguidade, quando o homem começou a usar a natureza em seu favor. Estas plantas, pela sua riqueza química e farmacológica, têm sido estudadas no intuito de comprovar atividades atribuídas pela crença popular ou mesmo obter novos compostos ativos. Neste contexto, a família Clusiaceae que compreende quarenta e nove gêneros de plantas de ampla distribuição tem sido investigada. Entre eles está o gênero Garcinia, também conhecido como Rheedia, que é rico em uma considerável diversidade de compostos, tais como benzofenonas polipreniladas e flavonóides. Neste trabalho foram obtidos os extratos hexânico, acetato-etílico e etanólico do pericarpo dos frutos de Garcinia brasiliensis. Após avaliação da atividade leishmanicida in vitro, o extrato hexânico se mostrou o mais ativo. Desta forma, foi fracionado e dele obteve-se três benzofenonas, 7-epi-clusianona, garciniafenona e guttiferona-a, que foram avaliadas quanto as suas atividades leishmanicida e toxicidade. No intuito de determinar um possível mecanismo de ação, os extratos foram avaliados quanto a sua capacidade de inibir proteases isoladas de amastigotas, sendo o extrato acetato etílico o que apresentou melhor resultado. Assim, este foi fracionado e dele se obteve o biflavonóide fukugetina, que também foi avaliado quanto à capacidade de inibir proteases, apresentando resultados muito satisfatórios. Todavia, quando o extrato acetato etílico e a fukugetina foram avaliados quanto à atividade leishmanicida in vitro não foi observado o mesmo efeito. Com estes resultados podemos observar que a polaridade dos extratos e das moléculas testados neste estudo pode ser um fator imprescindível na atividade leishmanicida, possivelmente por dificultar o transporte destas substâncias através de membrana plasmática. / The plants have been used in the treatment and cure of illnesses since the antique. When the men started to use the nature in your service. These plants, due to their chemical and pharmaceutical characteristics, have been studied in order to prove their theoretical medicinal capabilities or to obtain new active components. In this context, the Clusiaceae family is made up of forty nine species that are widely distributed, have been studied. Among these species is the genus Rheedia, also known as Garcinia, which is rich in a wide range of components, such as polyprenylated benzophenones and flavonoids. In this work were obtained the hexanic, ethyl-acetate and ethanolic extracts from pericarp of fruits from Garcinia brasiliensis. After the evaluation of leishmanicidal activity in vitro, the hexanic extract shown the more active, thus, this was fractioned yielding three benzophenones - 7-epi-clusianone, garciniaphenone and guttiferone-a – that were evaluated how their leishmanicidal and toxicity activities. With the intention of to determine a possible mechanism of action, these components were then evaluated of their capacity of to inhibit proteases isolated from amastigote forms of Leishmania, have been the ethyl-acetate extract the more effective. Thus this was fractioned yielding the biflavonoid fukugetin, witch also was evaluated in relation your proteases inhibition capacity, showing results very satisfactory. However, when the ethyl-acetate extract and the fukugetin were evaluated in relation the leishmanicidal activity in vitro, not was find the same effect. With these results, we might realize that the hydrophilic of extracts and molecules evaluated in this study can be an essential factor in the leishmanicidal activity, possible by to make transport difficult of this compounds across the plasmatic membrane. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

Pectinas são polímeros compostos principalmente por unidades de α-D-ácidos galacturônicos ligados por ligações α-1,4 presente na parede celular das plantas. De acordo com os compostos ramificados na cadeia principal pode-se dividir a pectina em vários tipos. Esses polissacarídeos possuem estruturas complexas, são solúveis em água e geralmente apresentam baixa toxicidade e possuem várias aplicações na indústria química, alimentícia e farmacêutica. Este trabalho teve como objetivo extrair, isolar e caracterizar os compostos pécticos presentes no epicarpo dos frutos de Garcinia brasiliensis (bacupari). Utilizou-se dois métodos de extração: água em meio neutro e água em meio ácido, ambos com aquecimento. Utilizou-se uma pectina comercial de citrus para comparação com as análises das pectinas do bacupari. A pectina foi obtida dos resíduos de epicarpo oriundos de extrações rotineiras e exaustivas com etanol, realizadas no laboratório de Fitoquímica e Química Medicinal (LFQM). O processo de extração de pectina em meio aquoso neutro, resultou em um rendimento de 7,48%, e o processo de extração em fase aquosa ácida resultou em um rendimento de 6,15%. A pectina dos frutos de G. brasiliensis, extraída com solução aquosa neutra, apresentou alto teor de metoxilação e esterificação e também apresentou maior teor de compostos fenólicos com maior capacidade sequestrante de radical livre DPPH do que as demais pectinas utilizadas. Além disso, foi avaliada a atividade antimicrobiana e antiinflamatória. As pectinas extraídas do bacupari e a comercial não apresentaram atividade antimicrobiana significativa quando avaliadas contra fungos e bactérias padrões Americam Type Culture Collection (ATCC), porém demonstraram atividade antiinflamatória considerável, quando comparadas ao padrão indometacina, testado no modelo de edema de pata de camundongos induzido por carragenina. Foram realizadas a caracterização estrutural, bem como a correlação com os dados de atividade biológica dos compostos naturais e semi-sintéticos (acetilados) conduzindo ao desenvolvimento de novos agentes antiinflamatórios a partir dos resíduos de extração dos frutos do bacupari. / Pectins are polymers mainly composed by unities of α-D galacturonic acids bonded by α- 1,4 links that are present in the plants cell wall. According to the branched composts in the main chain, a pectin can be divided in many kinds. Those polysaccharides have many complex structures, that are soluble in water and usually present low toxicity and have many uses in the chemical industry. This work aimed to extract, isolate and characterize the pectin composts present in the epicarps from the Garcinia brasilensis fruits, also known as bacupari. Two extraction methods were perfomed: one with water in a neutral solution and other with water in an acid one, both of them were heated. A comercial pectin from citrus was used to compare the analysis from the bacupari’s pectins. The pectin was obtained from residues of epicarp derived of routine and exaustive extractions with ethanol, that were realized at the Medical Phytochemistry and Chemistry Laboratory (MFCL). The process of pectin extraction in neutral aqueous solution resulted in a yield of 7,48% and the extraction process in the aqueous acid solution resulted in a yield of 6,15%. The pectin extracted from the g. brasiliensis fruit presented a high level of methoxylation and esterification and also showed higher phenolic content with more sequestring capacity of the DPPH free radical. Besides that, the antimicrobial and anti-inflammatory actvity were evaluated. The pectins extracted from the bacupari and the commercial one didn’t show a meaningfully activity agaisnt fungi and American Type Culture Collection (ATCC) standard bacterias, however they showed a considerable anti-inflammatory activity when compared to the indomethacin standard, which was tested on the carrageenan-induced paw edema in mices. Structural characterization, as well as data correlation of biological activity of natural and semi-sinthetic (acetylated) composts were performed, conducing to the development of new anti-inflammatory agents from the residues of the extraction of bacupari’s fruits.