Thesis (Ph. D.)--University of Canterbury, 2007. / Typescript (photocopy). "September 2007." Includes bibliographical references (p. 147-179). Also available via the World Wide Web.

El propósito del presente trabajo fue el evaluar la actividad antiinflamatoria de extractos de la especie Buddleja coriácea R. (Kiswara), además de su combinación con el fármaco antiinflamatorio dexametasona, mediante modelos preclínicos. Porque esta especie es empleada en la medicina tradicional para patologías inflamatorias. Inicialmente se determino la presencia de compuestos fenólicos en los extractos: Hidroalcohólico y Diclorometano/Hidroalcohólico de Buddleja coriácea R., mediante pruebas cualitativas, donde se identificaron terpenos, alcaloides, saponinas, taninos y flavonoides presentes en las hojas de la especie. Posteriormente se evaluó el efecto de inhibición de inflamación en el modelo de edema auricular a dosis de 3g/Kg de los extractos Hidroalcohólico y Diclorometano/Hidroalcohólico, además de su interacción con dexametasona donde se obtuvieron 21.39% y 31.02% de inhibición respectivamente, mientras que la dexametasona presento un 63.35% de inhibición pero cuando interacciona con los extractos presenta 45.64% y 68.08% de inhibición respectivamente, siendo estos estadísticamente significativos con respecto al control (p 0.05).

No / To analyze the fatty acid contents of Buddleja asiatica Lour, both the non-volatile oil and fat obtained from the n-hexane soluble sub-fraction were subjected to GC/MS using BSTFA (N,O-bis(trimethylsilyl) trifloroacetamide) derivatization. The oil showed the presence of six fatty acids including palmitic acid (46.75 %), linoleic acid (37.80 %), stearic acid (10.98 %), arachidic acid, margaric acid and lignoceric acid (< 3 %). Analysis of the fat revealed nine fatty acids including lignoceric acid (43.12 %), behenic acid (26.39 %), arachidic acid (9.29 %) and stearic acid (5.3 %). Cerotic acid, montanic acid, melissic acid and palmitic acid were found in low amounts (< 5 %) while trycosylic acid (4.83 %) was the only fatty acid with odd number of carbon atoms. The oil showed a low thermal stability. / Higher Education Commission Pakistan

Lignocellulosic bioethanol is currently being explored as a substitution to fossil fuels. Many lignocellulosic materials are being examined but the importance is to find those with attractive agro-energy features. Producing lignocellulosic ethanol is challenging because lignocellulosic biomass is resistant to chemical and biological degradation. To reduce biomass recalcitrance, a pretreatment stage is required. Pretreatment is considered to be the most intensive operating/operating cost component of cellulosic ethanol production. Therefore, research is heavily focused on understanding the effect of pretreatment technologies on the fundamental characteristics of lignocellulosic biomass. The first study in the thesis investigates Buddleja davidii as a potential biomass source for bioethanol production. The work focuses on the determination of ash, extractives, lignin, hemicellulose, and cellulose content in this plant, as well as detailed elucidation of the chemical structures of both lignin and cellulose by NMR spectroscopy. The study showed that B. davidii has several unique agro-energy features as well as some undesired characteristics. The second study presents research on the ethanol organosolv pretreatment (EOP) of B. davidii and its ability to produce enzymatically hydrolysable substrates. It was concluded that the removal of hemicellulose, delignification, reduction in the degree of polymerization (DP) of cellulose, and the conversion of crystalline cellulose dimorphs (Iα/Iβ) to the easily degradable para-crystalline and amorphous celluloses were the characteristics accounted for efficient enzymatic deconstruction of B. davidii after EOP. The third study provides a detailed elucidation of the chemical structure of ethanol organosolv lignin (EOL) of B. davidii by NMR spectroscopy. Such research was needed to understand the pretreatment mechanism in the context of delignification and alteration of the lignin structure. Future applications of the resulted EOL will be valuable for industrially viable bioethanol production process. EOP mainly cleaved β-O-4' interlinkages via homolysis, decreased the DP of lignin, and increased the degree of condensation of lignin. EOL had low oxygen content, molecular weight, and aliphatic OH as well as high phenolic OH, which are qualities that make it suitable for different co-product applications. The last study provides information on the anatomical characteristics of pretreated B. davidii biomass after EOP. The importance of this research was to further understand the alterations that occur to the cellular structure of the biomass which can then be correlated with its enzymatic digestibility. The results concluded that the physical distribution of lignin within the biomass matrix and the partial removal of middle lamella lignin were key factors influencing enzymatic hydrolysis.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The present work describes the phytochemical study and biological activity evaluation of the Buddleja brasiliensis Jacq. ex. Spreng and Artemisia verlotorum Lamotte. Were isolated three compounds from Buddleja brasiliensis: verbascoside (1), β-sitosterol (54) and stigmasterol (55), and it was carried out a phytochemical screening, a study of the ability to inhibit the enzymes prolyl oligopeptidase (POP), dipeptidyl peptidase (DPP IV) and acetylcholinesterase (AChE), a study of the antimicrobial activity of the specie and the extraction of the essential oil. The fractions F4 and F12 inhibited significantly POP and AchE activity. All Buddleja brasiliensis fractions tested presented low activity of DPP IV inhibition. The phenylpropanoid verbascoside (1) demonstrated a significant and selective inhibitory activity against POP (IC50 = 1.3 mM), showed weak activity against DPP-IV (IC50 >> 150 mM) and was inactive against AChE (pMIQ 9.6). The essential oil obtained from this species has as major constituents the a-tujaplicin monoterpene (11.5%) and the diterpenoid heneicosane (14.5%). This oil was inactive against all tested enzymes. The antimicrobial activity of 1, the crude extract and fractions of Buddleja brasiliensis were tested against several fungi and bacteria. Fractions dichloromethane and ethyl acetate showed moderate activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis, Streptococcus pyogenes, while 1 showed no antimicrobial activity. The crude extract, fractions and essential oil obtained from Artemisia verlotorum were also evaluated for inhibitory activity against the POP and DPP IV, as well as against various fungi and bacteria. Dichloromethane fraction of Artemisia verlotorum showed the best antimicrobial activity, mainly against the yeast Saccharomyces cerevisiae, with an MIC of 125 μg/mL. Analysis by gas chromatography coupled with mass spectrometry of the essential oil of Artemisia verlotorum indicated as the main components santonilil acetate (37.2%) and a-cadinol (9.59%). / O presente trabalho descreve o estudo fitoquímico e a avaliação da atividade biológica das espécies Buddleja brasiliensis Jacq. ex. Spreng e Artemisia verlotorum Lamotte. De Buddleja brasiliensis foram isolados três compostos: verbascosídeo (1), β-sitosterol (54) e estigmasterol (55). Desta espécie, fez-se a prospecção fitoquímica, a avaliação da capacidade de inibição das enzimas prolil oligopeptidase (POP), dipeptidil peptidase (DPP IV) e acetilcolinesterase (AChE), a obtenção do óleo essencial, e avaliação da atividade antimicrobiana. Nos testes de inibição frente à enzima POP foi observada significativa atividade das frações F4 e F12. Estas frações também demonstraram atividade inibitória da enzima AChE. Nos testes com a DPP IV verificou-se baixa atividade para todas as frações testadas. O fenilpropanóide verbascosídeo (1) demonstrou ter uma significativa e seletiva atividade inibitória frente à POP (IC50 = 1,3 μM), enquanto frente à DPP IV demonstrou fraca atividade inibitória (IC50 >>150 μM) e frente à AChE foi inativo (pMIQ de 9,6). O óleo essencial obtido desta espécie tem como constituintes principais o monoterpeno a-tujaplicina (11,5 %) e o diterpeno heneicosano (14,5 %). Este óleo foi inativo frente a todas as enzimas testadas. A atividade antimicrobiana de 1, do extrato bruto e das frações de Buddleja brasiliensis foi testada frente a vários fungos e bactérias. O composto 1 não apresentou nenhuma atividade antimicrobiana, enquanto que as frações que demonstram atividade moderada foram as frações Diclorometano e Acetato de etila para Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis, Streptococcus pyogenes. O extrato bruto, as frações e o óleo essencial obtidos de Artemisia verlotorum também foram avaliados quanto a atividade de inibição frente à POP e a DPP IV, bem como frente a vários fungos e bactérias. A fração Diclorometano de Artemisia verlotorum demonstrou a melhor atividade antimicrobiana, principalmente frente ao fungo Saccharomyces cerevisae, com uma CIM de 125 μg/mL. A análise por cromatografia gasosa acoplada a espectrômetro de massa do óleo essencial de Artemisia verlotorum indicou como constituintes principais acetato de santonilila (37,2 %) e a-cadinol (9,59%).

The Scrophulariaceae family is composed of some 25 genera and more than 1200 species. The genus Buddleja, widely distributed around the world, belongs to this family presently. Buddleja thyrsoides Lam. species, object of study in this work, is distributed in the southern of Brazil region and is popularly known by barbasco and cambarazinho-do-campo. Flowers and leaves decoction are used to treat bronchitis, asthma and cough. The first step of this was the morphoanatomical study of B. thyrsoides leaves aiming to established the identity of the species. The leaves present lanceolate-linear shape, straight adaxial face, serrata to subentire margin and little revolute close to the basis. The epidermis is uniseriate, with the epidermal cells of the adaxial face bigger than those of abaxial face, stomata and trichomes in abaxial face. The mesophyll is heterogeneous, bifacial and presents brachiblasts and crystals in druse form. The vascular system is of the open collateral type. Phytochemical screening showed alkaloids, flavonoids, cumarins, organic acids, antocyanic heterosides, saponins and volatile oils. The antioxidant activities, total phenolic contents, antimicrobial activities, antiplatelet action, acetylcholinesterase enzyme inhibition and toxicity of the crude extract (70% ethanol macerate) and dichloromethane, ethyl acetate and butanolic fractions of Buddleja thyrsoides were investigated. Total phenolic contents varied from 214.07 to 438.4 mg / g. The crude extract and fractions in order of increasing polarity exhibited scavenging activity with IC50 equal to 186.04, 137.70, 146.89, 165.71 μg/mL. A positive correlation between antioxidant activity and total phenolic contents was observed, where the ethyl acetate and dichloromethane fractions had the best results in both tests. The butanolic fraction was effective against S. cerevisiae (62.5 μg / mL for MIC and MFC) in the assessment of antimicrobial activity. The dichloromethane and ethyl acetate fractions were effective against S. aureus (250 and 500 μ g / mL for CIM and CBM), respectively. The crude extract and the fractions of B. thyrsoides did not shown toxicity against Artemia salina. The fractions (in order of increasing polarity) and crude extract showed antiplatelet action (42.35, 40.68, 43.57 and 41.52% respectively), whereas the AAS afforded 58.36% of inhibition. The crude extract inhibited in 22.77% the acetylcholinesterase enzyme after 24 hours. The major components of the essential oil were germacreno D (27.16%), 1,10-di - epi-cubenol (13.37%) and α - Cadinol (12.95%), biciclogermacreno (9.00%), δ - cadineno (8.59%) and cariofileno E (5.26%). / A família Scrophulariaceae é composta por cerca de 25 gêneros e mais de 1200 espécies. O gênero Buddleja, amplamente distribuído pelo mundo, atualmente, pertence a esta família. A espécie Buddleja thyrsoides Lam., objeto de estudo neste trabalho, encontra-se distribuída na região sul do Brasil e é conhecida popularmente por barbasco e cambarazinho-docampo. A decocção das flores e folhas é utilizada para bronquite, asma e tosse. Este trabalho objetiva em identificar morfoanatomicamente as folhas de B. thyrsoides, bem como analisar alguns parâmetros farmacológicos e biológicos para esta espécie. As folhas apresentam formato linear-lanceolado, face adaxial lisa, margem serreada até subinteira e pouco revoluta próxima à base. A epiderme é uniseriada, com as células epidérmicas da face adaxial maiores do que na face abaxial, estômatos e tricomas na face abaxial. O mesofilo é heterogêneo, bifacial e apresenta braquiblastos e cristais na forma de drusas. O sistema vascular é do tipo colateral aberto. Na análise fitoquímica foram detectados alcalóides, flavonóides, cumarinas, ácidos orgânicos, heterosídeos antociânicos, saponinas e óleos voláteis. O extrato bruto (de um macerado etanólico a 70 %) e frações diclorometano, acetato de etila e butanólica de Buddleja thyrsoides foram investigados quanto à atividade antioxidante, conteúdo de fenólicos totais, atividade antimicrobiana, ação antiagregante, inibição da enzima acetilcolinesterase e quanto à toxicidade. O conteúdo de fenólicos totais variou de 214,07 a 438,4 mg/g. O CI50 da atividade antioxidande para o extrato bruto e frações em ordem crescente de polaridade (186,04; 137,70; 146,89; 165,71 μg/mL). Foi observada uma correlação positiva entre a atividade antioxidante e o conteúdo de fenólicos totais, onde as frações acetato de etila e diclorometano apresentaram os melhores resultados em ambas análises. Na avaliação da atividade antimicrobiana, a fração butanólica foi efetiva contra S. cerevisae (62,5 μg/mL para CIM e CFM). As frações diclorometano e acetato de etila foram eficazes contra o S. aureus (250 e 500 μg/mL para CIM e CBM), respectivamente. O extrato bruto e as frações de B. thyrsoides não apresentaram toxicidade frente Artemia salina. As frações (em ordem crescente de polaridade) e extrato bruto mostraram ação antiagregante plaquetária (42,35; 40,68; 43,57 e 41,52 %, respectivamente), sendo que o AAS apresentou 58,36 % de inibição. O extrato bruto inibiu em 22,77 % a enzima acetilcolinesterase após 24 horas. Os componentes majoritários do óleo essencial são Germacreno D (27,16%), 1,10-di-epi-Cubenol (13,37 %) e α-Cadinol (12,95 %), Biciclogermacreno (9,00 %), δ-cadineno (8,59 % ) e Cariofileno E (5,26%).

No / Buddlejol (1), a new sterol, has been isolated from the ethyl acetate soluble fraction of the antispasmodic plant Buddleja asiatica along with stigmasterol (2), lignoceric acid (3), taraxerol (4) and alpha-amyrin (5), respectively. The structure of Buddlejol (1) was established as (24S)-stigmast-5,22-diene-7 beta-ethoxy-3 beta-ol by spectral analysis and comparison with closely related structures. Buddlejol revealed to be a competitive inhibitor of chymotrypsin with the Ki value of 10.60 A mu M as indicated by Lineweaver-Burk and Dixon plots and their re-plots against its chymotrypsin inhibition assay, while the other compounds showed less inhibitory potential. The bioassay-guided isolation was stimulated by the preliminary cytotoxic screening of various fractions of B. asiatica.