Triquentrinas A são produtos naturais marinhos com atividade biológica e alta complexidade estrutural. Estes fatores tornam estes alcaloides e compostos análogos, como os herbindóis, alvos para a síntese total e plataforma para o desenvolvimento de novas metodologias sintéticas. Nesta Dissertação de Mestrado, procurou-se completar a síntese estereosseletiva da (+)-cis-triquentrina A empregando-se intermediários sintéticos protegidos com o grupo benzila que poderão ser usados para a futura prospecção de novos compostos com atividade biológica. A rota proposta baseia-se na obtenção do ácido (S)-3-(1-benzil-4-etil-1H-indol-7-il)butanóico, um intermediário-chave já descrito por Silva e colaboradores no percurso da síntese total da (+)-trans-triquentrina A, e da finalização da síntese de acordo com a abordagem proposta pelo grupo de RajanBabu para conversão do análogo protegido com grupo tosila à (+)-cis-triquentrina A. A resolução enzimática do intermediário-chave com lipase de Pseudomonas cepacia imobilizada em terra diatomácea foi otimizada, resultando em rendimentos de 32% e 99% ee na metade do tempo descrito anteriormente. Na etapa-chave da síntese, o (S)-ácido foi submetido a uma acilação de Friedel-Crafts intramolecular na presença de anidrido trifluoroacético que produziu o produto de ciclização desejado com 40% de rendimento. Na etapa final da síntese, o intermediário sintético protegido com grupo benzila não pode ser convertido à (+)-cis-triquentrina A, empregando-se a metodologia desenvolvida para a redução do composto análogo tosilado, dada a diferença de reatividade imposta pela troca do grupo protetor. As etapas realizadas até o penúltimo intermediário sintético (S)-8, consta com 10 etapas e rendimento global de 1,3%. viii As diferenças eletrônicas e estruturais relacionadas a diferentes grupos protetores poderão ser refletidas em variações na atividade antiproliferativa de indóis relacionados a triquentrinas. Ainda no interesse de preparar moléculas para envio à análises de atividade antiproliferativa preparou-se um composto relacionado à síntese da trans-triquentrina A tendo como etapa-chave uma contração de anel mediada por I(III) em 21% de rendimento, cujo trabalho foi incluído nos anexos. / Trikentrins A are marine natural products with biological activity and high structural complexity. These factors make these alkaloids and analogous compounds, such as herbidoles, targets for total synthesis and platform for the development of new synthetic methodologies. In this Master\'s Dissertation, we attempted to complete the stereoselective synthesis of (+)-cis-trikentrin A using synthetic intermediates protected with the benzyl group that could be used for the future prospection of new compounds with biological activity. The proposed route is based on the preparation of (S)-3-(1-benzyl-4-ethyl-1H-indol-7-yl) butanoic acid, a key intermediate already described by Silva et al., in the course of total synthesis (+)-trans-triquentrin A, and the final part of the synthesis according to the approach proposed by the RajanBabus group for conversion of the protected analogue with tosyl group to (+)-cis-trikentrin A. The enzymatic resolution of the key intermediate with Pseudomonas cepacia lipase immobilized on diatomaceous earth was optimized, resulting in 32% yield and 99% ee in half the time described above. In the key step of the synthesis, the (S)-acid was subjected to an intramolecular Friedel-Crafts acylation in the presence of trifluoroacetic anhydride which yielded the desired cyclization product in 40% yield. In the final step of the synthesis, the synthetic intermediate protected with benzyl group couldnt be converted to (+)-cis-trikentrin A, employing the methodology developed for the reduction of the tosylated analogous compound, given the difference of reactivity imposed by exchange of the protective group. The steps carried out up to the synthetic intermediate (S)-8, consists of 10 steps and overall yield of 1.3%. Electronic and structural differences related to different protective groups may be reflected in variations in the antiproliferative activity of indoles related to trikentrins. A compound related to the synthesis of trans-triquentrin A having as its key step an I(III) mediated ring contraction in 21% yield was also prepared in the interest of preparing molecules for antiproliferative activity analysis, whose work was included in the appendix.

Prior work using a contractility bioassay determined that the serotonin (5-HT) receptor subtype 5-HT2A is present in bovine lateral saphenous veins and plays a role in ergot alkaloid-induced vascular contraction in steers grazing endophyte-infected (Epichloë coenophiala) tall fescue (Lolium arundinaceum). A study was conducted to determine what 5-HT receptors are involved in vasoconstriction of bovine gut vasculature. The findings of this study indicate that 5-HT2A is present and may play a role in ergot alkaloid induced vasoconstriction. A second study was conducted to determine if ergot alkaloids were transported in the small intestine. The active transporter, peptide transporter 1 (PepT1), was evaluated for its role in the transport of various concentrations of ergot alkaloids across Caco-2 cell monolayers. Results indicate that CEPH, ERT, EXT, and LSA do move across Caco-2 cell monolayers, but appear to utilize PepT1 at larger concentrations. Overall, the demonstrated presence of 5-HT2A receptors in the bovine gut vasculature established a potential for vascular interference by ergot alkaloids entering the bloodstream through transepithelial absorption.

L’objectif de ce travail a consisté en le développement de nouveaux processus multi-composant radicalaires et leur application en synthèse organique. Des carbo-alcénylation d'oléfines ont ainsi été réalisées avec de nouveaux précurseurs de radicaux, des oléfines diverses, en présence de Z-diphénylsulfonyléthylène comme accepteur terminal. Les conditions de la réaction ont été optimisées, en introduisant notamment la diphénylsulfonylhydrazine comme amorceur de radicaux sous irradiation UV, et substitut au couteux DTBHN. Des conditions sans étain ont également été étudiées avec l’objectif de remplacer le réactif (Bu3Sn)2 par des radicaux silylés. Le tris (triméthylsilyl)silylthiopropene a ainsi été testé avec succès en tant qu'agent de propagation des chaînes radicalaires. A l’issue de ce travail méthodologique, nous avons développé une stratégie de synthèse de l'Eucophylline, un alcaloïde isolé de Leuconotis griffithii, dont le squelette tétracyclique a été élaboré sur la base d’une réaction de carbo-oximation radicalaire d’oléfine. Ce processus multicomposant, suivi d’une réduction de la fonction oxime et d’une lactamisation offre une voie d’accès rapide au motif bicyclo[3.3.1]lactame, intermédiaire-clé de la synthèse. Une réaction de type Friedländer entre ce lactame et un ortho-aminobenzonitrile a permis d’accéder au squelette tetrahydrobenzo[1,8]naphthyridine de l'Eucophylline. La synthèse du composé modèle a enfin été complétée par l’introduction du substituant vinylique par un couplage de Heck. / The aim of this work was to develop new radical multi-component processes and their application in organic synthesis. Carbo-alkenylation processes were thus performed with new radical precursors, different olefins, in the presence of Z-diphenylsulfonylethylene as a terminal acceptor. Reaction conditions have also been optimized, including the diphenylsulfonylhydrazine as a radical initiator under U.V. irradiation, and substitute to the costly DTBHN. Tin-free conditions were also screened with the goal of replacing (Bu3Sn)2 with silyl radicals. Tris(trimethylsilyl)silylthiopropene was tested with success as a radical chain carrier. After this methodology studies, we developed a strategy toward the synthesis of Eucophylline, an alcaloid isolated from Leuconotis griffithii, which tetracyclic skeleton was elaborated based on a carbo-oximation of olefin. This multicomponent process, followed by a reduction of the oxime function and a lactamization offered a fast access to the bicyclo[3.3.1]lactam, a key-intermediate in the synthesis. A Friedländer-type reaction between this lactam and an ortho-aminobenzonitrile allowed an access to the Eucophylline tetrahydrobenzo[1,8]naphthyridine skeleton. The synthesis of the model compound was finally completed with the introduction of the vinylic substituent through a Heck coupling.

Chapter one; “(-)-Rhazinilam and (-)-Rhazinal: Alkaloids with Anti-mitotic Properties Derived from Kopsia teoi”, provides the background information behind the motives that initiated this research project. The plant alkaloid (-)-rhazinilam [(-)-1] and its naturally-occurring derivative (-)-rhazinal [(-)-13] both exhibit potent anti-mitotic activities and, as such, are interesting targets for total synthesis. Chapter one is a review of the literature regarding these two compounds and discusses the occurrence, proposed biosynthetic origins, structural elucidation and biological activites of compound (-)-1 and that of its analogues including alkaloid (-)-13. Previous total syntheses of these two compounds are then examined, concluding with the only reported total synthesis of compound (-)-13. Developed within the Banwell research group, this total synthesis produced the racemic modification of alkaloid (-)-13 due to a lack of any stereocontrol in the key intramolecular Michael addition step. This unprecedented key step, involving cyclisation of the C2 of pyrrole onto an N-tethered and ?,?-disubstituted acrylate to produce a quaternary-carbon stereogenic centre, would be of greatly enhanced utility if it could be achieved in a catalytic-enantioselective fashion. The realisation of this goal is the central aim of the research conducted within this thesis. ¶ Chapter two; “Investigating Asymmetric Induction in the Intramolecular Michael Addition of pyrrole to N-Tethered Acrylates and Related Species”, introduces the model study used to direct research towards achieving the goal of asymmetric induction in the title process. The model is a somewhat simplified version of the original process used in the total synthesis of compound (-)-13 involving cyclisation of the C2 of pyrrole onto an N-tethered and ?-monosubstituted Michael acceptor, to produce a tertiary-carbon stereogenic centre. This simplification allows the rapid synthesis of a broad range of potential substrates for use in the title process, thus enabling the investigation of various different approaches to inducing asymmetry therein. High levels of asymmetric induction are observed with the use of chiral substrates or catalysts, facilitating the synthesis of both 6- and 7-membered rings annulated to pyrrole with construction of the relevant tertiary-carbon stereogenic centre in enantio-enriched form. For the reactions producing a 6-membered ring annulated to pyrrole, unambiguous proof of the absolute sense of asymmetric induction observed in the intramolecular Michael addition event is established using a chemical correlation study involving elaboration of a key indolizine-type cyclisation product, to the plant alkaloid of known absolute stereochemistry, (-)-tashiromine [(-)-75]. For the reaction producing a 7-membered ring annulated to pyrrole, the same information is obtained via X-ray crystallographic analyses of a dibrominated derivative of a key pyrroloazepine-type cyclisation product. ¶ Chapter three “An Enantioselective Total Synthesis of the Alkaloid (-)-Rhazinal: An Anti-mitotic Agent Isolated from Kopsia teoi.”, focuses on the application of methodology developed in the previous chapter, to the original goal of inducing asymmetry in the intramolecular Michael addition reaction, involving cyclisation of the C2 of pyrrole onto an N-tethered and ?,?-disubstituted acrylate to produce a quaternary-carbon stereogenic centre. This is ultimately achieved in a catalytic-enantioselective fashion, resulting in the first such total synthesis of the anti-mitotic alkaloid (-)-rhazinal [(-)-13]. ¶ Chapter four “Extending the Reaction Manifold to the Syntheses of Related Natural Products: A Formal Total Synthesis of (+)-Aspidospermidine and Syntheses of (-)-Rhazinilam and (-)-Leuconolam from (-)-Rhazinal”, describes three extensions to the reaction manifold used in the enantioselective total synthesis of alkaloid (-)-13: The acquisition in an enantioselective manner, of an intermediate previously obtained in racemic form, en route to the racemic modification of the natural product (±)-aspidospermidine [(±)-134], constitutes a formal and enantioselective total synthesis of (+)-aspidospermidine [(+)-134]. The direct deformylation of (-)-rhazinal [(-)-13], is carried out, to produce the parent alkaloid (-)-rhazinilam [(-)-1]. The pyrrole ring present in (-)-rhazinilam [(-)-1] is oxidised, to produce the related natural product (-)-Leuconolam [(-)-12] which has not, hitherto, been prepared by total synthesis. ¶Chapter five contains the experimental procedures and characterisation data associated with compounds described in chapters two to four.

Salvia divinorum is a hallucinogenic sage used to treat illness by the Mazatec Indians of Mexico. Salvinorin A (1a), a neoclerodane diterpenoid isolated from the plant, is a potent, selective agonist at the kappa opioid receptor (KOR), and is the first non-nitrogenous opioid. The plant is used recreationally as a hallucinogen, but is unpopular due to its dysphoric effects. 1a has been prohibited in Australia under an invalid systematic name. An early report of psychoactive alkaloids in S. divinorum proved to be irreproducible. Similarly, tests in mice suggesting the presence of psychoactive compounds other than 1a were confounded and therefore unreliable. In this work, an improved isolation method for 1a was developed, using filtration through activated carbon to decolourise the crude extract. Six new diterpenoids were isolated: salvinorins D–F (1d–1f) and divinatorins A–C (28a–28c). Five known terpenoids not previously reported from this species were also isolated. The structure–activity relationships of 1a were evaluated via selective modifications of each functional group. Useful synthetic methods are reviewed, including the first thorough review of furanolactone hydrogenations. Testing of the derivatives at the KOR suggests that the methyl ester and furan ring of 1a are required for activity, but that the lactone and ketone functionalities are not. Other compounds from S. divinorum did not bind to the KOR, suggesting that 1a is the plant’s active principle.

Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2016-09-01T17:22:47Z No. of bitstreams: 1 Tese Sandra Virginia A. Hohlemwerger.pdf: 5499646 bytes, checksum: 10ad62e0bf160a9a5e0326cba05adb7f (MD5) / Approved for entry into archive by Vanessa Reis (vanessa.jamile@ufba.br) on 2016-09-02T16:27:00Z (GMT) No. of bitstreams: 1 Tese Sandra Virginia A. Hohlemwerger.pdf: 5499646 bytes, checksum: 10ad62e0bf160a9a5e0326cba05adb7f (MD5) / Made available in DSpace on 2016-09-02T16:27:00Z (GMT). No. of bitstreams: 1 Tese Sandra Virginia A. Hohlemwerger.pdf: 5499646 bytes, checksum: 10ad62e0bf160a9a5e0326cba05adb7f (MD5) / Este trabalho tem como objetivo contribuir com o conhecimento fitoquímico das espécies do gênero Zanthoxylum. O presente estudo justifica-se devido ao escasso número de informações sobre a composição química de espécies deste gênero, endêmicas do Brasil, já que espécies pertencem a família Rutaceae vem sendo utilizadas na medicina popular em todo o mundo, inclusive no Brasil, e têm sido apontadas como potencial fonte para protótipos ou novos fármacos. A literatura correlaciona algumas atividades biológicas destas espécies com a presença de alcalóides, dentre eles os alcalóides benzilisoquinolínicos os quais além de serem relacionados com a atividade farmacológica são apontados como possíveis marcadores quimiossistemáticos. As espécies Zanthoxylum rhoifolium, Z. stelligerum e Z. tingoassuiba foram coletadas no semi-árido baiano e submetidas a procedimentos fitoquímicos convencionais os quais levaram a identificação dos alcalóides benzo[c]fenantridinicos (diidroqueleritina), (angolina), (arnotianamida) e (pseudo-norqueleritrina), do alcalóide protoberberínico (cis-N-metilcanadina iodeto) e do alcalóide pseudoprotoberberínico (2,3-metilenodioxi 10,11-dimetoxi tetrahidroprotoberberina iodeto) além disso também foi isolado o alcalóide aporfínico (predicentrina-metil-iodeto) e o alcalóide (metil antranilato de N-metila); as furanocumarinas (imperatorina) (xantotoxina) (isopipinelina) sendo ainda revelada a presença da lignana (senamina) e dos triterpenos (lupeol), (-amirina) e (amirinona), estes dois últimos encontrados apenas na cera epicuticular das folhas de Z. tingoassuiba. O óleo volátil das folhas de Z. tingoassuiba foi obtido pela extração com CO2 super crítico e através da hidrodestilação de onde foram identificados 06 monoterpenos, 10 sesquiterpenos e o alcalóide (metil antranilato de N-metila) este óleo apresentou atividade antifúngica, antibacteriana e capacidade de associação com lipossomas. As identificações destas substâncias foram efetuadas com base nos seus espectros de IV, RMN 1H e 13C uni e bidimensionais, cromatografia gasosa acoplada a espectrômetro de massas, espectrometria de massas por inserção direta e comparações com dados da literatura. Os metabólitos isolados e identificados nas espécies estudadas são condizentes com os já encontrados no gênero Zanthoxylum, como os alcalóides quaternários benzilisoquinolínicos e as furanocumarinas os quais estão associados ao caráter primitivo do gênero. Os resultados promissores das atividades biológicas, antifúngica, antibacteriana, antioxidante e antiparasitária, corroboram para a classificação deste gênero como fonte para o desenvolvimento de novos fármaco / This work aims to improve the knowledge phytochemical for the species of the genus Zanthoxylum. This study is justified due to the scarcity of information about the chemical composition of species of this genus, endemic to Brazil. Species belonging to family Rutaceae has been used in folk medicine around the world, including Brazil, have been identified as a potential source for prototypes or new drugs. The literature correlates the biological activities of some of these species with the presence of alkaloids, among them alkaloids benzylisoquinolines which besides being related to the pharmacological activity are considered possible chemical markers. Species Zanthoxylum rhoifolium, Z. stelligerum and Z. tingoassuiba were collected in semi- arid environments and subjected to conventional procedures phytochemicals which led to the identification of benzo [c] phenanthridine alkaloids (dihydrochelerythrine), (angoline), (arnottianamide) and (pseudo-norchelerythrine), and the protoberberínico alkaloid (cis-N-methylcanadine iodide) and pseudoprotoberberine alkaloid (2,3- methylendioxy 10,11-dimethoxy tetrahydroprotoberberine iodide) was also isolated the aporphine alkaloid (predicentrine-methiodide) and alkaloid (methyl N-methyl anthranilate); the furocumarins (imperatorin) (xanthotoxin) (isopimpinellin) besides the furofuran lignan (sesamin) and the triterpenoids (lupeol), (-amyrin) and (amyrinone) the latter two found only in the wax epicuticular on the leaves of Z. tingoassuiba. The volatile oil from leaves of Z. tingoassuiba was obtained by supercritical CO2 and by hydrodistillation of which were identified 06 monoterpenes, 10 sesquiterpenes and the alkaloid (methyl N-methyl anthranilate) this oil showed antifungal and antibacterial activity and ability to loaded into multilamellar liposomes. The identification of these substances was based on their IR spectra, 1H and 13C NMR single and two-dimensional, gas chromatography-mass spectrometry, mass spectrometry direct insertion, and comparison with literature data. The metabolites isolated and identified in the three species are consistent with those already found in the genus Zanthoxylum, such as quaternary benzylisoquinolines alkaloids and furnocumarinas which are associated with the primitive character of the genus. The promising results of biological activities, antifungal, antibacterial, antioxidant and anti- interference, collaborate with the classification of genus as a source for the development of new drugs.

Made available in DSpace on 2016-06-02T20:36:48Z (GMT). No. of bitstreams: 1 5580.pdf: 12897921 bytes, checksum: 7de4b9e632d7de714eb77bf5afad7e21 (MD5) Previous issue date: 2013-03-27 / Universidade Federal de Sao Carlos / The leaf-cutting ants are social species, living in permanent colonies. The genera of the greater importance, Atta (saúvas) and Acromyrmex (quenquéns), are the major plagues of Brazilian agriculture, to cause serious damages by its large and uncontrolled grazing, which leads to serious economic losses for many agrosystems. In the same way, the corn maize weevil, Sitophilus zeamais, belonging to the family Curculionidae, is the major pest species of stored corn. The corn grains are drilled and damaged by larvae and adults, causing numerous qualitative and quantitative losses of corn grains disabling its marketing. As the consequence, the effective control of these plagues is extremely important for agriculture and economy that aims to provide the nacional and international market quality products with high commercial value. In this context, the phytochemistry study of Annona sylvatica provides an alternative way to control these plagues, since that studies of these species showed a wide variety of natural products with promising activities, including, insecticide and fungicide, due to the large presence of acetogenins and lignans as secondary metabolites. This study allowed the isolation of fourteen compounds (four lignans, four acetogenins, three alkaloids and three steroids), however were identified eleven compounds due to the high structural complexity presented for the acetogenins. The identification techniques employed have been using single and two-dimensional spectra of 1H and 13C NMR and Mass Spectrometry (HPLC/NMR; HPLC/ESI-MS/MS e HPLC/APCI-MS/MS). For the biological tests, the ethanolic extract of the seeds stood out as potentially toxic to three bioassays cited above. The leaves and branches dichlorometanic fractions obtained by liquid-liquid partitioning of the extracts were toxic for leaf-cutting ants and their symbiotic fungus. Among the acetogenins isolated, only one of them showed a significant activity against corn maize weevil. / As formigas cortadeiras são espécies sociais, vivendo em colônias permanentes. Os gêneros de maior importância econômica, Atta (saúvas) e Acromyrmex (quenquéns), são as principais pragas da agricultura brasileira, por causarem sérios danos pela sua grande e descontrolada herbivoria, o que gera sérios prejuízos econômicos para muitos agroecossitemas. Nessa mesma vertente, o gorgulho do milho, Sitophilus zeamais, pertencente à família Curculionídea, é a principal espécie praga do milho armazenado. Os grãos são perfurados e danificados pelas larvas e adultos, inutilizando sua comercialização. Como consequência, o controle eficaz de tais pragas é de extrema importância para a agricultura e economia do país que visa oferecer ao mercado nacional e internacional produtos de qualidade e alto valor comercial. Nesse contexto, o estudo fitoquímico de Annona sylvatica fornece uma forma alternativa de controle a estas pragas, uma vez que estudos destas espécies mostraram uma ampla variedade de produtos naturais com atividades promissoras, entre elas, inseticida e fungicida, devido à ampla presença de acetogeninas e lignanas como metabólitos secundários. Este estudo resultou no isolamento de 14 compostos (quatro lignanas, quatro acetogeninas, três alcaloides e três esteroides), no entanto, foram identificados 11 compostos, devido à alta complexidade estrutural apresentada para as acetogeninas. As técnicas de identificação utilizadas foram através dos espectros uni e bidimensionais de RMN 1H e 13C e por Espectrometria de Massas (HPLC/NMR; HPLC/ESI-MS/MS e HPLC/APCI-MS/MS). Para os ensaios biológicos, o extrato etanólico das sementes destacou-se como potencial tóxico frente aos três bioensaios citados acima. As frações diclorometânicas das folhas e ramos obtidas através do particionamento líquido-líquido dos extratos foram tóxicas para as formigas cortadeiras e seu fungo simbionte. Dentre as acetogeninas isoladas, apenas uma apresentou uma atividade significativa frente ao gorgulho do milho.

Submitted by Cristhiane Guerra (cristhiane.guerra@gmail.com) on 2017-02-03T15:37:52Z No. of bitstreams: 1 arquivototal.pdf: 4718514 bytes, checksum: 73b76bdede325ded91073c36318f98f2 (MD5) / Made available in DSpace on 2017-02-03T15:37:52Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4718514 bytes, checksum: 73b76bdede325ded91073c36318f98f2 (MD5) Previous issue date: 2016-02-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The synthetic alkaloid 2-methoxy-4-(7-methoxy-1,2,3,4- tetrahydroisoquinolin-1-yl) phenol encoded as MHTP, has a 93.45% yield and it had no genotoxic effect with low acute toxicity in preclinical analyses, with LD50 higher than 1000 mg / kg. The MHTP presented vasorelaxant and anti-inflammatory effects in acute inflammation models by orally administration. The aim of this study was to evaluate the immunomodulatory effect of MHTP, administered intranasally (in) in experimental allergic pulmonary inflammation induced by ovalbumin (OVA) by means of allergic (measurement of serum IgE) and inflammatory parameters (cell migration and production of cytokines, the fluid broncoalveolar- washed BAL, mucus production and histopathological pulmonary remodeling). Female BALB / c mice were sensitized with OVA at days 0 and 12 of the experimental protocol and on days 19 and 22 were treated via i.n. MHTP with the doses of 2.5 and 5 mg / kg or dexamethasone (2 mg/kg). After each treatment was aerosol challenged with OVA for 30 minutes daily; 24 hours after the last challenge, it was collected biological material for evaluation of allergic lung inflammation characteristic on allergic asthma. Treatment with MHTP at dose of 2.5 mg/kg decreased (p<0,05) the migration of total lymphocytes, (p<0,0001) the lymphocytes CD4+ and production of the cytokines IL-13, IL-4, IL-17 and IL-10. The dose of 5 mg/kg, in turn, decreased (p <0,05) the production of OVA-specific IgE and migration of lymphocytes CD3+; (p<0,0001) migration of leukocytes, total lymphocytes CD4+, macrophages and (p<0,001) eosinophils; (p<0,0001) the percentage of granulocytes; the production of the cytokines IL-13, IL-4, IL-17 and IL- 10. In addition, MHTP reduced inflammatory histopathological parameters such as hyperplasia and hypertrophy of goblet cells and mucus overproduction. The results infer that the immunomodulatory mechanism of MHTP is related to regulation of Th2 profile that is responsible for generating and maintaining allergic pulmonary inflammation process characteristic of allergic asthma. / O alcaloide sintético 2-methoxy-4-(7-methoxy-1,2,3,4- tetrahydroisoquinolin-1-yl) phenol, codificado como MHTP, possui rendimento de 93,45%, não apresentou efeito genotóxico com baixa toxicidade aguda pré clínica e DL50 maior que 1.000 mg/kg. O MHTP apresentou efeito vasorelaxante e antiinflamatorio em modelo de inflamação aguda, administrado por via oral. O objetivo do presente trabalho foi avaliar o efeito imunomodulador do MHTP, administrado por instilação nasal (i.n.), na inflamação pulmonar alérgica experimental induzida por ovalbumina (OVA), por meio dos parâmetros alérgico (quantificação de IgE sérica), inflamatório (migração celular, produção de citocinas, no fluido do lavado broncoalveolar- BAL) e morfológico (produção de muco e histologia pulmonar geral). Camundongos fêmeas BALB/c foram sensibilizados com OVA nos dias 0 e 12 do protocolo experimental e nos dias 19 a 22 foram tratados por via i.n. com MHTP nas doses 2,5 ou 5 mg/kg ou Dexametasona (2 mg/kg). Após cada tratamento, foi realizado os desafios por aerossol com OVA, durante 30 minutos diários; 24 horas após o último desafio, foi coletado o material biológico necessário para avaliação da inflamação pulmonar alérgica, característica da asma alérgica. O tratamento com MHTP na dose de 2,5 mg/kg diminuiu (p<0,05) a migração de linfócitos totais, (p<0,0001) linfócitos CD4+ e a produção das citocinas IL-13, IL-4, IL-17 e IL-10. A dose de 5 mg/kg, por sua vez, diminuiu (p<0,05) a produção de IgE-OVA específica e migração de linfócitos CD3+; (p<0,0001) migração de leucócitos totais, linfócitos CD4+, macrófagos e (p<0,001) eosinófilos; (p<0,0001) o percentual de granulócitos e a produção das citocinas IL-13, IL-4, IL-17 e IL-10. Além disso, MHTP reduziu os parâmetros inflamatórios histopatológicos e morfológicos e a hiperprodução de muco. Os resultados obtidos inferem que o mecanismo imunomodulador do MHTP está relacionado à regulação do perfil Th2, que é responsável por gerar e manter o processo inflamatório alérgico pulmonar, característico da asma alérgica.

Submitted by Vasti Diniz (vastijpa@hotmail.com) on 2017-09-05T12:23:50Z No. of bitstreams: 1 arquivototal.pdf: 1922103 bytes, checksum: a207d4c95bf749dd37ae0eb1728c7e80 (MD5) / Made available in DSpace on 2017-09-05T12:23:50Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1922103 bytes, checksum: a207d4c95bf749dd37ae0eb1728c7e80 (MD5) Previous issue date: 2017-04-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Inflammation is an immune response that aims to establish a tissue homeostasis during an infection or injury. Recognized as a beneficial process, an inflammation can become harmful when in excess. Thus, therapeutic therapies that are applied to the resolution of the inflammation are studied and developed, with natural products and their derivatives playing a prominent role in the discoveries of new anti-inflammatory molecules. In this context, the objective of this study was to evaluate the antiinflammatory potential of milonine, a Cissampelos sympodialis Eichl alkaloid, in vivo, against the release of mediators by using the murine model of acute inflammation and pain. Swiss mice pretreated with milonine were submitted to protocols of: paw edema induced by LPS, prostaglandin (PGE2), bradykinin (BK) and serotonin (5-HT) to evaluate the anti-edematogenic activity of the alkaloid; microvascular permeability induced by acetic acid to obtain a total protein concentration and histological analysis of the peritoneum; the models of carrageenan-induced peritonitis to evaluate the effect of milonine on the migration of the total and differential cells; and the formalin test to evaluate its nociceptive activity. The results show that milonine was capable of significantly reduce (p<0.001) the formation of the paw edema induced by LPS, PGE2, BK, however it was not capable to reduce the edema induced by 5-HT; significantly reduced (p<0.05) the extravazation of liquid to the peritoneum induced by acetic acid, maintaining its morphology preserved. The alkaloid was also able to inhibit (p<0.01) the migration of the total leukocytes to the peritoneal cavity during the carrageenan-induced inflammation, decreasing the number of polymorphonuclear cells (PMN) without changing mononuclear cells (MNs). In the formalin test, the paw licking time of the animals was inhibited (p<0.01) only in the second phase through the intraperitoneal (i.p.) administration, corroborating the inhibition of inflammatory mediators. Therefore, this study demonstrates that milonine has an antiinflammatory and analgesic activity due to the inhibition of the action of mediators essential for the start of the inflammatory process. / O processo inflamatório é uma resposta benéfica do organismo frente a uma lesão ou infecção, visando a eliminação da injúria inicial, bem como as consequências dessa injúria. Entretanto, a inflamação, em resposta exagerada, pode ser prejudicial. Dessa forma, se faz necessário o desenvolvimento de novos fármacos com propriedades anti-inflamatória e analgésica, sendo utilizado como estratégia terapêutica a busca por moléculas oriundas de produtos naturais que possuam tais propriedades. Nesse contexto, o objetivo desse estudo foi avaliar o potencial anti-inflamatório da milonina, alcaloide de Cissampelos sympodialis Eichl frente à liberação de mediadores inflamatórios utilizando o modelo murino de inflamação aguda e dor. Camundongos Swiss pré-tratados com milonina foram submetidos aos protocolos de: edema de pata induzido por LPS, prostaglandina (PGE2), bradicinina (BK) e serotonina (5-HT) para avaliar a atividade antiedematogênica do alcaloide; permeabilidade microvascular induzida por ácido acético para obter a concentração de proteínas totais e análise histológica do peritônio; os modelos de peritonite induzida por carragenina para avaliar o efeito da milonina sobre a migração de células totais e diferenciais; e o teste da formalina para avaliar a sua atividade nociceptiva. Os resultados demonstraram que a milonina foi capaz de reduzir significativamente (p<0,001) a formação do edema de pata induzido por LPS, PGE2, BK, porém não foi capaz de reduzir o edema induzido pela 5-HT; diminuiu significativamente (p<0,05) o extravasamento de líquido para o peritônio induzido por ácido acético, mantendo a sua morfologia preservada. O alcaloide também foi capaz de inibir (p<0,01) a migração de leucócitos totais para a cavidade peritoneal durante a inflamação induzida por carragenina, diminuindo o número de células polimorfonucleares (PMN) sem alterar células mononucleares (MNs). No teste da formalina, o tempo de lambida da pata dos animais foi inibida (p<0,01) por milonina apenas na segunda fase quando administrada por via intraperitoneal (i.p.). Portanto, este estudo demonstrou que a milonina possui atividade anti-inflamatória e analgésica por inibir a ação de mediadores essenciais para o início do processo inflamatório.

Made available in DSpace on 2015-05-14T12:59:54Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 7265191 bytes, checksum: 432cedb6fda0226806b02fd0634dfff5 (MD5) Previous issue date: 2013-08-12 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The poisonous plants to livestock research has been restricted only the identification of poison species, epidemiology and clinical signs. But the active principle of toxic plants are little known and its knowledge its very important to develop preventive methods to poisoning which are responsible by countless cattle deaths. This work aimed the contribution to knowledge of active principle these plants, Crotalaria vitellina Ker Gawl (Fabaceae) e Ipomoea philomega (Vell.) House (Convolvulaceae). This work aimed the contribution to knowledge of active principle these plants, Crotalaria and Ipomoea were submitted to phytochemical study for the isolation of its chemical constituents by cromatographic methods followed by its identification through spectroscopic techniques such as Infrared (IR), one and two-dimentional Nuclear Magnetic Resonance (NMR) of 1H and 13C, and Mass Spectrometry (MS) besides literature data. The phytochemical study of C. vitellina resulted on the isolation of the pyrrolizidine alkaloid (type otonecine) named crotavitelin (Cv-1), and were obtained from the crude extract of its fruits, described by first time in the literature. This substance was subjected to acute toxicological evaluation according to OECD Guide 423 (Guideline for Testing of Chemicals), in mice (males and females) orally exposed to 50 and 300 mg/Kg doses and showed a low toxicity on the parameters evaluated. However, histopatologic studies should be performed to investigate the possible toxic effects in celular and tissue levels. Ipomoea philomega was submitted also to phytochemical studies and were isolated eight compounds from the dicloromethane phase of the ethanolic crude extract of the leaves: lanosterol (Ip-1), caffeic acid (Ip-2), ethyl p-coumarate (Ip-3), lupeol (Ip-4), ethyl caffeate (Ip-5), umbelliferone (Ip-6), scopoletin (Ip-7), and the 1,2-benzopirone (Ip-8), has been described for first time in I. philomega. / A pesquisa sobre plantas tóxicas para animais têm-se limitado principalmente à identificação das espécies, bem como à sua epidemiologia, patologia e sinais clínicos. Sendo, no entanto, pouco conhecidos os seus princípios ativos, cujo conhecimento é de grande importância no desenvolvimento de métodos preventivos da intoxicação, responsáveis por inúmeras mortes de animais e, consequentemente, perdas econômicas. Visando contribuir para o conhecimento dos princípios ativos dessas plantas, as espécies Crotalaria vitellina Ker Gawl (Fabaceae) e Ipomoea philomega (Vell.) House (Convolvulaceae) foram submetidas a um estudo fitoquímico para isolamento de seus constituintes químicos por métodos cromatográficos, seguidos de identificação através de métodos espectroscópicos tais como Infravermelho (IV), Ressonância Magnética Nuclear (RMN) de 1H e 13C uni e bi-dimensionais e Espectroscopia de Massas (EM) juntamente com a comparação com dados da literatura. O estudo fitoquimico do extrato etanólico bruto dos frutos e folhas de C. vitellina resultou no isolamento de um alcalóide pirrolizidínico do tipo otonecina, descrito pela primeira vez na literatura, nomeado crotavitelina (Cv-1). Essa substância foi submetida avaliação da toxicidade pré-clínica aguda, de acordo com o Guia da OECD-423 (Guideline for Testing of Chemicals), em camundongos (machos e fêmeas) nas doses de 50 e 300 mg/Kg e apresentou baixa toxicidade nos parâmetros avaliados. Entretanto, estudos histopatológicos, especialmente em nível de tecido hepático, devem ser realizados para a investigação de possíveis efeitos tóxicos em nível celular e tecidual. Ipomoea philomega, Convolvulaceae, igualmente submetida a estudo fitoquímico do extrato etanólico bruto das suas folhas possibilitou o isolamento de oito substâncias da fase diclorometano: lanosterol (Ip-1), ácido cafeico (Ip-2), p-cumarato de etila (Ip-3), lupeol (Ip-4), cafeato de etila (Ip-5), umbeliferona (Ip-6), escopoletina (Ip-7) e a 1,2- benzopirona (Ip-8), descritas pela primeira vez para I. philomega.