Thesis (doctoral)--Rijksuniversiteit te Utrecht, 1983. / Includes bibliographical references.

Medicinal plants have provided important advances in the treatment of numerous diseases and many plant-derived drugs are currently in use or under investigation for the treatment of many ailments including cancer.<p>A phytochemical analysis of the methanol extract from the seed of <i>Saponaria vaccaria</i> L. cultivated in Saskatchewan was performed which resulted in the detection of several bisdesmosidic saponins. A high-performance liquid chromatographic method using photodiode array and single quadrupole electrospray mass detection for analysis and profiling was developed. Due to their structural similarities, purification of bisdesmosidic saponins was challenging. However, monodesmosidic saponin Vaccaroside B and cyclopeptides Segetalin A, Segetalin B, and a new cyclopeptide, segetalin I [whose structure was proposed to be cyclo(Gly1-Pro2-Tyr3-Tyr4-Pro5-Phe6)], were purified employing various chromatographic techniques such as HPLC, VLC, PTLC). <p>Crude methanol extracts of <i>S. vaccaria</i> seed were evaluated for cytotoxic activity using the methyl-thiazol-tetrazolium non-radioactive cell proliferation assay (MTT assay). Various concentrations of the extract (2-50 ug/mL) were tested against a series of four human cancer cell lines (WiDr, colon; MDA-MB-231, breast; NCI-417, lung and PC-3, prostate). The human foreskin (BJ)-derived normal human fibroblast cell line CRL-2522 was included as a non-cancerous control. Results showed that cytotoxic activities from the seed extract were greater than commercially available Quillaja saponaria saponin. <p>The human cancer cell lines were also exposed to fractions containing high titers of saponins as well as semi-purified saponins (ca. 80%). All bisdesmosidic saponins and fractions thereof showed cytotoxicity against the cell lines studied. The effect was particularly strong in breast and prostate cancer cell lines with IC50 values in the range 14 ug/mL. Monodesmosidic saponins, phenolics and cyclopeptides did not show activity even at the highest concentration (50 ug/mL) tested in this study. Chemical modifications of the saponin structures resulted in an overall decrease in activity, but an increase in selectivity in comparison to CRL-2522. Time and concentration-dependent studies using the nuclear stains propidium iodide and Hoechst 33342, demonstrated that the stimulation of apoptosis was the mechanism of cytotoxic action. When breast and prostate cell lines were exposed to small amounts (4-7 mM) of bisdesmosidic saponins Segetalin H (MW 1448) and Segetalin I (MW 1464), it was observed that apoptosis was induced at an early incubation time (4-10 h). Activation of caspases and changes in membrane potential were determined by flow cytometry.<p>As a result of this study, we propose that triterpene bisdesmosidic saponins from the seed of <i>Saponaria vaccaria</i> L. represent a new type of drug having potential antitumor/anticancer activity due to their ability to induce apoptosis in vitro in human cancer cell lines at low concentrations. These compounds are extracted from a plant that can be easily cultivated using conventional agricultural equipment in Western Canada.

Medicinal plants have provided important advances in the treatment of numerous diseases and many plant-derived drugs are currently in use or under investigation for the treatment of many ailments including cancer.<p>A phytochemical analysis of the methanol extract from the seed of <i>Saponaria vaccaria</i> L. cultivated in Saskatchewan was performed which resulted in the detection of several bisdesmosidic saponins. A high-performance liquid chromatographic method using photodiode array and single quadrupole electrospray mass detection for analysis and profiling was developed. Due to their structural similarities, purification of bisdesmosidic saponins was challenging. However, monodesmosidic saponin Vaccaroside B and cyclopeptides Segetalin A, Segetalin B, and a new cyclopeptide, segetalin I [whose structure was proposed to be cyclo(Gly1-Pro2-Tyr3-Tyr4-Pro5-Phe6)], were purified employing various chromatographic techniques such as HPLC, VLC, PTLC). <p>Crude methanol extracts of <i>S. vaccaria</i> seed were evaluated for cytotoxic activity using the methyl-thiazol-tetrazolium non-radioactive cell proliferation assay (MTT assay). Various concentrations of the extract (2-50 ug/mL) were tested against a series of four human cancer cell lines (WiDr, colon; MDA-MB-231, breast; NCI-417, lung and PC-3, prostate). The human foreskin (BJ)-derived normal human fibroblast cell line CRL-2522 was included as a non-cancerous control. Results showed that cytotoxic activities from the seed extract were greater than commercially available Quillaja saponaria saponin. <p>The human cancer cell lines were also exposed to fractions containing high titers of saponins as well as semi-purified saponins (ca. 80%). All bisdesmosidic saponins and fractions thereof showed cytotoxicity against the cell lines studied. The effect was particularly strong in breast and prostate cancer cell lines with IC50 values in the range 14 ug/mL. Monodesmosidic saponins, phenolics and cyclopeptides did not show activity even at the highest concentration (50 ug/mL) tested in this study. Chemical modifications of the saponin structures resulted in an overall decrease in activity, but an increase in selectivity in comparison to CRL-2522. Time and concentration-dependent studies using the nuclear stains propidium iodide and Hoechst 33342, demonstrated that the stimulation of apoptosis was the mechanism of cytotoxic action. When breast and prostate cell lines were exposed to small amounts (4-7 mM) of bisdesmosidic saponins Segetalin H (MW 1448) and Segetalin I (MW 1464), it was observed that apoptosis was induced at an early incubation time (4-10 h). Activation of caspases and changes in membrane potential were determined by flow cytometry.<p>As a result of this study, we propose that triterpene bisdesmosidic saponins from the seed of <i>Saponaria vaccaria</i> L. represent a new type of drug having potential antitumor/anticancer activity due to their ability to induce apoptosis in vitro in human cancer cell lines at low concentrations. These compounds are extracted from a plant that can be easily cultivated using conventional agricultural equipment in Western Canada.

Universiẗat, Diss., 2004--Frankfurt (Main).

Universiẗat, Diss., 2005--Düsseldorf. / Erscheinungsjahr an der Haupttitelstelle: 2004.

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2017-04-27T12:32:10Z No. of bitstreams: 1 2003 - Patricia Miranda da Costa.pdf: 2556665 bytes, checksum: 84eaee321ba021876a1fd855d0767602 (MD5) / Made available in DSpace on 2017-04-27T12:32:10Z (GMT). No. of bitstreams: 1 2003 - Patricia Miranda da Costa.pdf: 2556665 bytes, checksum: 84eaee321ba021876a1fd855d0767602 (MD5) Previous issue date: 2003-11-28 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Solvent partition and chromatographic fractionation of metanolic extract from the leaves and wood of L. arianeae lead to the isolation of triterpenes, one carboidrate, saponin and flavonoid. 3b,6b,19a -trihydroxiursa-12-en- 28-oic acid, 3b,6b,24-trihydroxiursa-12-en- 28-oic acid, 3,5,7-trihydroxi-4?-metoxi,6-flavone-( sulfonate), 3b-hydroxiolean-12-en-28- ?ico acid, 3b-O-b-D-galactopiranosyl-(6?-1 para hydroxi benzoil)-ursa-12-en-28-oic acid, 1-metyl glycopiranosyl and mixture of two flavonoids. The 3,5,7-trihydroxi-4?-metoxi,6- flavone-(sulfonate) and 3b-O-b-D-galactopiranosyl-(6?-1 para hydroxi benzoil)-ursa-12-en- 28-oic acid are being descricted for the first time in the literature. The IR, 1H NMR, 13C NMR and MS spectra analysis was used for structural determination. / O fracionamento dos extratos das folhas e madeira de L. arianeae atrav?s de processo de parti??o com solventes e t?cnicas cromatogr?ficas conduziu ao isolamento dos ?cidos 3b,6b, 19a -triidroxiursan-12-eno-28-?ico, 3b,6b,24,19a-tetraidroxiursan-12-eno-28-?ico, 3b-hidroxiolean-12-eno-28-?ico, da 3,5,7-triidroxi, 4?-metoxi, 6-sulfonato, flavona, da 3b- O-b-D-galactopiranosil-(6?-p-hidroxi-benzo?la)-ursan-12-eno-28-?ico, da 1-metil glicose, e da mistura de flavon?ides. As estruturas das subst?ncias foram deduzidas atrav?s da an?lise dos espectros de IV, RMN de 1H e 13C, incluindo experimentos 2D e espectro de massas das subst?ncias naturais e dos derivados. Este ? o primeiro registro destes constituintes no g?nero Licania. A flavona e saponina est?o sendo descritos pela primeira vez na literatura. As folhas e as cascas da esp?cie E. longipes foram submetidas a extra??o com solventes org?nicos e os extratos foram fracionados atrav?s de parti??o e t?cnicas cromatogr?ficas. As fra??es reunidas foram submetidas a t?cnicas cromatogr?ficas e cristaliza??o. Esses processamentos conduziram ao isolamento dos ?cidos 1a,2b,3a,19a -tetraidroxiursan-12- eno-28-?ico, da saponina 3b-O-b-D-glicopiranosil-sitosterol e dos triterpenos fridelinol e 3b,24-diidroxifridelano. As estruturas das subst?ncias foram deduzidas atrav?s das t?cnicas citadas acima. Este ? o primeiro registro desta saponina no g?nero Eschweilera. O triterpeno 3b,24-diidroxifridelano foi descrito pela primeira vez na literatura (COSTA, P.M. & Carvalho, M. G., Annais da Academia Brasileira de Ci?ncias, 2003).

The goal of this PhD research was directed to the isolation and structural elucidation of the secondary metabolites from four species of Durio plants, especially from those growing in the island of Borneo; This island contains most Durio species. The structures of isolated compounds were deduced using state-of-the-art approaches, including 1D and 2D nuclear magnetic resonance (NMR), mass spectrometric (MS) and circular dichroism (CD) analyses. From the bark of the woody species Durio zibethinus Murr. (Bombacaceae), that was collected in Pontianak, a chemically diverse group of oxygenated compounds was obtained. These included caffeoyl triterpenes, lignans and phenylpropanoid derivatives. During this research, one new compound 27-O-cis-caffeoylcylicodiscate was obtained which resulted from a trans to cis isomerization; this process might be caused by acid and light. Two presumed fungal metabolites were also isolated during this study, these possibly arising from fungal (unknown species) contamination of the bark. The relative stereochemistry of 4-hydroxymellein were also established by conversion to it diacetate and measurement of the coupling constant between H-3 and H-4. The absolute configuration was determined by comparison of []D values with those from the literature. Assignment of the 13C NMR spectrum for (+)-(R)-de-O-methyllasiodiplodin was completed using HSQC-TOCSY data. The methanol extract of the bark of Durio kutejensis (Hassk.) Becc. was fractionated and purified by C18-HPLC. Two new caffeoyl pentacyclic triterpenes 3-O-trans-caffeoyl-2-hydroxyolean-12-en-28-oic acid and 3-O-trans-caffeoyl-2-hydroxyurs-12-en-28-oic acid, together with five known compounds were isolated. The known compound fraxidin had previously been obtained from D. zibethinus Murr. The bark of Durio carinatus Mast., a plant species which has non edible fruit was collected in Sambas district, West Kalimantan. Three new lignans, boehmenan X, (-)-(7S,8S)-threo-carolignan X and (-)-(7R,8S)-erythro-carolignan X, and a new caffeoyl triterpene 3-O-cis-caffeoyl betulinic acid, together with three known lignans and one known caffeoyl triterpene, were obtained from the crude MeOH extract. The relative configurations and conformational models of the diastereomers threo- and erythro-carolignan X were determined by the coupling constant data for H-7′ and H-8′ and supported by 2D NMR spectroscopic analysis including NOESY and HSQC-HECADE in CDCl3. However, the magnitude of the coupling constants for both compounds in MeOH-d4 was similar. The stability of the individual conformers was solvent dependent and must be considered for their contribution to the overall conformational equilibria. This stability was determined by steric interactions, dipole repulsion effects and intramolecular hydrogen bonding. The absolute configuration of the two new diastereomers was established by application of circular dichroism (CD) and optical rotation []D measurements. Another Durio plant studied and collected in Bengkayang region, West Kalimantan, was Durio oxleyanus Griff. From the MeOH extract of its bark, two new lignans, (-)-(7S,8S)-threo-carolignan Y and (-)-(7R,8S)-erythro-carolignan Y were isolated using C18-HPLC. The relative stereochemistry of the carolignan Y diastereomers could not be deduced from the coupling constants for H-7′ and H-8′ either in CDCl3 and MeOH-d4, because of the presence of multiple conformers. The conformational model for the diastereomers of carolignan Y was similar to that of threo- and erythro-carolignan X in MeOH-d4. Using 2D NMR spectroscopic methods, including NOESY and HSQC-HECADE, the relative stereochemistry of the threo and erythro isomers of carolignan Y was confirmed. Further, the absolute configuration of threo- and erythro-carolignan Y was explored via analysis of CD spectra and by comparison of experimentally obtained []D values with literature values.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Universidade Federal do Cearà / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / The known metabolites of the genus Senna have a variety of applications, while the oldest is like purging and preparation is present in the British Pharmacopoeia, European, American and Brazilian indicated for cases of constipation. Are used in folk medicine preparations of various species of the genus as, stomachic, febrifuge, antianemics, antiblenorrÃgica, antinefrÃtica,antidot a, antimycotic, diuretic, fungicidal, laxative and against skin diseases. In this stdy is described the investigation of chemical of Senna georgica H. S. Irwin Barneby (Leguminoseae), popularly known as "Lava pratoâ and "mata pasto". The literature review revealed the complete absence of studies chemical, biological and pharmacological performed with this species, which is why the plant was selected for the study. The chemical study started with chromatographic treatment of hexane andethanol extracts of Senna which allowed the isolation and identification of two anthraquinones crisofanol and fisciona,the triterpene lupeol, two phytosteroid shaped and acid free and glycosylated triacontanÃico. Another stage of the work was devoted to the pharmacological study of S.georgica, which analyzed by testing the antioxidant potential, cytotoxicity and anticolinasterÃsico of extracts hexane and ethanol of flowers, pods, leaves, stem and root of the plant. The structural determinations were made using techniques of infrared spectroscopy, nuclear magnetic resonance of a 1-H-and C-13 uni-dimensional, mass spectrometry and comparison with data reported in the literature. / Os metabÃlitos conhecidos do gÃnero Senna apresentam uma variedade de aplicaÃÃes, embora a mais antiga seja como purgativa e sua preparaÃÃo està presente na FarmacopÃia britÃnica, europÃia, americana e brasileira com indicaÃÃo para casos de constipaÃÃo. Na medicina popular sÃo utilizadas preparaÃÃes de vÃrias espÃcies do gÃnero como tÃnica, estomÃquica, febrÃfuga, anti-herpÃtica, antianÃmica, antiblenorrÃgica, antinefrÃtica, antÃdota, antimicÃtica, diurÃtica, parasiticida, laxante, e contra doenÃas de pele. Este trabalho descreve a investigaÃÃo quÃmica de Senna georgica H. S. Irwin & Barneby (Leguminoseae), popularmente conhecida como âmata pastoâ e âlava pratoâ. O levantamento bibliogrÃfico revelou a completa ausÃncia de estudos quÃmicos, biolÃgicos e farmacolÃgicos realizados com esta espÃcie, motivo pela qual selecionou-se a planta para o estudo. O estudo quÃmico iniciou-se com o tratamento cromatogrÃfico dos extratos hexÃnico e etanÃlico do caule de S. georgica o que possibilitou o isolamento e identificaÃÃo das antraquinonas crisofanol e fisciona, o triterpeno de esqueleto lupano, o lupeol, dois fitoesteroides em forma livre e glicosilados e o Ãcido triacontanÃico. Outra etapa do trabalho foi dedicada ao estudo farmacolÃgico de S. georgica, que atravÃs de ensaios analisou o potencial antioxidante, citotoxidade e anticolinasterÃsico dos extratos etanÃlico e hexÃnico das flores, vagens, folhas, caule e raiz da planta. As determinaÃÃes estruturais foram realizadas atravÃs das tÃcnicas espectromÃtricas de infravermelho, ressonÃncia magnÃtica nuclear de hidrogÃnio-1 e carbono-13 uni e bidimensionais, espectrometria de massas e comparaÃÃes com dados relatados na literatura.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Este trabalho relata o estudo quÃmico do extrato hexÃnico e etanÃlico do caule de Senna cana H.S Irwin & Barneb, popularmente conhecida no Nordeste como candieiro preto e SÃo JoÃo, coletada em MucugÃ-BA. O estudo do Ãleo fixo do caule de Senna. cana resultou na identificaÃÃo de oito Ãcidos graxos, totalizando 83,13% da composiÃÃo do Ãleo fixo. Destacaram-se como componentes majoritÃrios os Ãcidos graxos dos Ãcidos saturados: palmÃtico (26,83%), lignocÃrico (18,60%) e o nonadecanÃico (11,18%). O tratamento cromatogrÃfico dos extratos hexÃnico e etanÃlico do caule da Senna cana, possibilitou o isolamento e caracterizaÃÃo de duas antraquinonas 1,8-diidroxiladas, crisofanol e fisciona, de um Ãcido carboxÃlico de cadeia longa denominado de Ãcido triacontanÃico e de dois triterpenos lupeol e friedelina, bem como a mistura &#946;-sitosterol e estigmasterol. O potencial farmacolÃgico e biolÃgico de Senna cana, foi investigado atravÃs de ensaios de atividade antioxidante, anticolinesterÃsica e larvicida. Das amostras analisadas, o extrato etanÃlico foi o que apresentou melhor resultado, com o I.V 39,61%. As determinaÃÃes estruturais foram realizadas atravÃs de anÃlises espectroscÃpicas, tais como: espectroscopia na regiÃo do infravermelho, de ressonÃncia magnÃtica nuclear de hidrogÃnio-1 e carbono-13, espectrometria de massas e comparaÃÃo com dados da literatura. / This paper reports the chemical study of the hexane and ethanol extracts from the Senna cana H.S Irwin & Barneby, called in the Northeast of Brazil as "Candieiro preto" and "SÃo JoÃo", collected in MucugÃ-BA. The study of the fixed oil of the stem of Senna cana resulted in the identification of eight fatty acids, amounting to 83.13% of fixed oil composition. Stood out as the major components of fatty saturated acids: palmitic (26.83%), lignoceric (18.60%) and nonadecanoic (11.18%). The chromatographic treatment of the hexane and ethanol extracts of the stem from the Senna cana, allowed the isolation and characterization of two dyhydroxylated 1,8-anthraquinones, chrysophanol and physcion, a long chain carboxylic acid named triacontanoic acid, two triterpenes called lupeol and friedelin and the mixtures &#946;-sitosterol and stigmasterol. The pharmacological and biological potencial from Senna cana was investigated by antioxidant activity, anticholinesterase and larvicide tests. The ethanol extract showed the best result with 39.61% IV. The structural determination was performed using spectroscopic analysis, such as infrared spectroscopy, hydrogen-1 and carbon-13 nuclear magnetic resonance, mass spectrometry and comparison with the results founded on the literature.

Oleanolic and ursolic acid (OA and UA) are triterpenic acids with diverse biological activities that are of interest to the pharmaceutical industry. To investigate the scope for producing these compound using cell suspension cultures of Salvia species, calli from S. officinalis, S. virgata and S. fruticosa were induced using several plant growth regulator (PGR) combinations. Eleven lines were selected for suspension induction from a pool of calli. Six suspension cultures were established successfully and cultivated in the Respiration Activity MOnitoring System® (RAMOS®) to obtain online data on their growth kinetics and to establish appropriate sampling schedules for the determination of their OA and UA production. Based on their observed growth behaviour, OA and UA contents, and aggregation properties, one suspension culture from each studied Salvia species was selected for further optimisation. The μmax values for these suspension cultures ranged from 0.20 to 0.37°d-1, their OA and UA contents were greater than 1.3 and 1.2 mg g-1, respectively, and they afforded maximum volumetric yields of 21.0 mg l-1 for OA and 32.8 mg l-1 for UA. These results will be useful in the development of a refined Salvia suspension-based process for OA and UA production.