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The objectives of the doctoral dissertation were achieved by determining the chemical composition of the extracts of H. physodes, isolation, structural determination and testing of the biological activity of physodalic acid, physodic acid, 3-hydroxyphysodic acid and atranorin. In this dissertation antioxidant activity, cytotoxic activity on cells of human cervix carcinoma and on rat thymocytes, anticholinesterase activity, antimicrobial activity and influence on the formation of micronuclei in lymphocytes of human blood plasma of physodalic acid, physodic acid, 3-hydroxyphysodic acid and atranorin were investigated The antioxidant activity and anticholinesterase activity were investigated by spectrophotometric methods. Cytotoxic activity on cells of human cervix carcinoma and rat thymocytes was assayed by a colorimetric MTT assay, and CCK-8 assay, respectively. The impact on the formation of micronuclei in lymphocytes of human blood plasma was investigated by micronucleus test. Antimicrobial activity was tested by using microdilution method. Comparing the literature available to us, and these results, it was found that for the first time was investigating the effect of Hypogymnia physodes methanol extract on cells of human cervical cancer and the number of micronuclei in lymphocytes of human blood plasma, impact of atranorin on the number of micronuclei in lymphocytes of human blood plasma and serum activity cholinesterase, antioxidant activity of 3-hydroxyphysodic acid and its effects on rat thymocytes, the number of micronuclei in lymphocytes of human blood plasma, the cells of the human cervical cancer and the activity of serum cholinesterase and the impact physodic acid on rat thymocytes, the number of micronuclei in human lymphocytes blood plasma, the cells of the human cervical cancer and serum cholinesterase activity and antioxidant activity of physodalic acid and its effect on rat thymocytes, the number of micronuclei in lymphocytes of human blood plasma, the cells of the human cervical cancer and the activity of serum cholinesterase. 3-Hydroxyphysodic acid shows a greater anti-oxidant activity from a commercial antioxidant 3,5-di-tert-butyl-4-hydroxytoluene (BHT), and reduce the number of micronucleus cells in human blood plasma 2.2 times more than the commercial protectant amifostine, recommending it as good candidate for in vivo experiments.

Ethyl acetate extracts of cultures grown in liquid Czapek and on solid rice media of the fungal endophyte Fusarium oxysporum SS46 isolated from the medicinal plant Smallanthus sonchifolius (Poepp.) H. Rob., Asteraceae, exhibited considerable cytotoxic activity when tested in vitro against human cancer cells. Chromatographic separation yielded anhydrofusarubin (1) and beauvericin (2) that were identified based on their ¹H and 13C NMR data. Compounds 1 and 2 showed the strongest cytotoxic activity against different cancer cell lines. Compound 2 also showed promising activity against Leishmania braziliensis. Hexanic extract of F. oxysporum SS50 grown on solid rice media also afforded a mixture of compounds that displayed cytotoxic activity against different cancer cell lines. Chemical analysis of the mixture of compounds, investigated by gas chromatography-mass spectrometry (GC-MS), showed that there was a predominance of methyl esters of fatty acids and alkanes.

The essential oil (EO) of aerial parts of Lantana camara L., Verbenaceae, from Simões, Piaui, Northeast of Brazil, were obtained by hydrodistillation and analyzed by GC-FID and GC-MS techniques. In total, 68 compounds were identified. The most representative compounds of the oil were mono and sesquiterpenes. The main compounds found in the oil of the leaves in different months were β-caryophyllene (10.5%, in June of 2009), sabinene (7.98%, in September of 2008), limonene (7.68%, in September of 2008), spathulenol (11.64%, in September of 2008). The oil from stems of L. camara was characterized by a largest amount of sesquiterpenoids, with spatulenol (15.9%) and caryophyllene oxide (17.1% in June of 2009), as main components. β-Gurjunene (32.7%, in September of 2008) was the most prominent compound in the stems oils, which was absent or at very low relative abundance in leaves. L. camara essential oils from leaves were cytotoxic to V79 mammalian cells and also to Artemia salina, showing 50% lethal concentration (LC50) values from 0.23 µg/mL. The in vitro data obtained in this study suggested that EO may also be effective treating yeast infection in patients infected with fluconazole and terbinafine resistant isolates, but its toxicity must be monitored carefully.

The syntheses of new Pt-II complexes with 4-acyl-5-pyrazolones [HQ(Ph) = 1-phenyl-3-methyl-4-benzoylpyrazol-5-one, a; HQ(py,CF3) = 1-(2-pyridyl)-3-methyl-trifluoroacetylpyrazol-5-one, b] are reported: trans-[PtCl2(DMSO)(HQ(Ph))] (1a), cis-[PtCl2(DMSO)(HQ(Ph))] (2a), [PtCl(DMSO)(Q(py,CF3))] (1b), and [PtCl2(KQ(py,CF3))] (2b). All complexes were characterized by multinuclear (H-1, C-13, F-19, and Pt-195), multidimensional NMR spectroscopy and single-crystal X-ray diffraction analyses. The isomer trans-[PtCl2(DMSO)(HQ(Ph))] (1a) crystallizes in two polymorphic forms, which correspond to two different conformers. In the trans-syn conformer (1a(M)) (monoclinic, space group P2(1)/c), the H atom of the enol group [O(1)H(1)] establishes an intramolecular hydrogen bond with the ketone oxygen (O2), whereas in the trans-anti conformer (1a(O)) (orthorhombic, space group Pca2(1)), the same groups are involved in an analogous intermolecular H bond. Interestingly, the cis isomer (2a) showed in solution and at room temperature slow rotation around both the Pt-N1 and the Pt-S bonds, which restrained both the acylpyrazolone and DMSO ligand motion. Moreover, the water solubility of the complexes trans- and cis-[PtCl2(DMSO)(HQ(Ph))] (1a and 2a) and [PtCl2(KQ(py,CF3))] (2b) allowed to perform in vitro biological assays on platinum-sensitive human endometrium (HeLa) and platinum-resistant human breast (MCF-7) cancer cell lines. It was also possible to compare their cytotoxicity to cisplatin. Interestingly, trans isomer 1a showed higher cytotoxicity on HeLa cells compared to the cis isomer 2a.

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Luis R??diger.pdf: 13443546 bytes, checksum: f45078022552e0a3713a149b282b0569 (MD5) Previous issue date: 2012-11-01 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior PHYTOCHEMICAL AND CITOTOXIC STUDY OF BURSERACEAE OLEORESINS. The Burseraceae family includes approximately 700 specimens distributed in 19 genres, and many of these specimens do not present phytochemical studies. Interested on describing the chemical composition, evaluate the cytotoxic activity and isolate substances of chemical and cytotoxic interest, this work has been divided in three parts: Determination of the extraction in hexane chemical profiles; cytotoxicity evaluation of oleoresins against HCT-8 (collon), MDA/MB-435 (breast) and SF-295 (glioblastom); and isolation of substances present in the oleoresins. The isolated substances were identified by mass spectrometry techniques (GC-MS and ESI-IT-MS) and nuclear magnetic resonance (NMR). In the volatile compounds profile it was possible to observe that among the monoterpenes, the p-cymene was detected more times, in 11 out 23 specimens evaluated. The highest concentration of this compound was observed in Protium apiculatum (2.4%), however the compound mostly found in this specimen is ??pinene (5.1%). Among the sesquiterpenes, the compound that presented highest concentration was the junenol, in P. cf. rubrum (23.9%) and P. cf. ferrugineum (11.3%) being isolated and elucidated. The samples with the most complex sesquiterpenic profile were P. tenuifolum and Tetragastris panamensis, with common detection of sesquiterpenes of bisabolane, cadinane, cubebane, eudesmane, guaiane and muurulane series. The total ion chromatograms evaluations showed the presence of ??- and ??-amyrin in all the samples, confirming the presence of these triterpenes as family markers. In P. cf. ferrugineum, it has been observed the absence of ??- and ??-amyrone, and it was the differential of this specimen among other samples. About the triterpenes, glutinol (first description in the genres studied) and the multiflorenol (first description in the family) were detected. The chemometric analysis resulted in five distinct cluster (C-1 to C-5), where C-1 presented high concentration of ??- and ??-amyrone and absence of brein and maniladiol; C-2, having potentially producers of new triterpene and volatile compound specimens; C-3 with medium concentration among the variables analyzed; C-4 with high concentration of brein and maniladiol; and C-5, having ??- and ??-amyrin high concentration specimens. Te cititoxicity assays showed that two extracts in hexane (P. cf. rubrum e Trattinnickia peruviana - HrTPE) showed inhibition against HCT-8 (93.08 % for both specimens) and SF-295 (93.46 e 86.34%, respectively) tumor cells lines. In the extracts in acetate and acetyl, inhibition for P. giganteum var giganteum and Trattinnickia glaziovii against HCT-8 (88.7 e 92.9%, respectively) and MDAM B-435 (97.8 e 96.2%, respectively) lines were observed. HrTPE was submitted to fractioning, being identified the ??-amyradienol. From the extract in ethyl acetate of P. paniculatum var. modestum, breine and maniladiol triterpenes in binary mixture, 3-hydroxy-tirucalla-8,24-diene-21-oic acid, pure and in binary mixture with 3-hydroxy-tirucalla-7,24-dieno-21-oic acid were isolated. In the P. bahianum ethyl acetate extract, the ursolic and oleanolic acids were isolated. From the Tetragastris panamensis ethyl acetate extract, an unpublished triterpene (3-oxo-triptocallic B acid) and two triterpenes decribed for the first time in the Burseraceae family (triptocallic B and 3-oxo-katononic acids) were isolated. A 2,3-seco-isobriononico acid biosynthesis from multiflorenol proposal was presented. The family chemosystematics has been discussed. A fam??lia Burseraceae compreende aproximadamente 700 esp??cies distribu??das em 19 g??neros, sendo que v??rias destas esp??cies n??o apresentam estudos fitoqu??micos. Com interesse em descrever a composi????o qu??mica, avaliar a atividade citot??xica e isolar subst??ncias de interesse qu??mico e citot??xico, o trabalho foi dividido em tr??s partes: determina????o do perfil qu??mico dos extratos em hexano; avalia????o da citotoxicidade de extratos das oleorresinas contra as linhagens HCT-8 (c??lon), MDA/MB-435 (melanoma) e SF-295 (glioblastoma); e isolamento de subst??ncias presentes nas oleorresinas. As subst??ncias isoladas foram identificadas pelas t??cnicas de espectrometria de massas (CG-EM e EMCIESI) e resson??ncia magn??tica nuclear (RMN). No perfil dos compostos vol??teis observou-se que dentre os monoterpenos, o p-cimeno foi detectado mais vezes, em 11 das 23 esp??cies avaliadas. A maior concentra????o deste composto foi observada em Protium apiculatum (2,4%), contudo o componente majorit??rio nesta esp??cie ?? o ??-pineno (5,1%). Dentre os sesquiterpenos o componente que apresentou maior concentra????o foi o junenol, em P. cf. rubrum (23,9%) e P. cf. ferrugineum (11,3%) sendo isolado e elucidado. As amostras com perfil sesquiterp??nico mais complexo foram P. tenuifolum e Tetragastris panamensis, com detec????o em comum de sesquiterpenos das s??ries bisabolano, cadinano, cubebano, eudesmano, guaiano e muurulano. As avalia????es dos cromatogramas de ??ons totais demonstraram a presen??a dos ??- e ??-amirina em todas as amostras, confirmando a presen??a destes triterpenos como marcadores desta fam??lia. Em P. cf. ferrugineum , observou-se a aus??ncia de ??- e ??-amirona, o que diferenciou esta esp??cie das demais amostras. Dentre os triterpenos, foram detectados o glutinol (primeira descri????o nos g??neros estudados) e o multiflorenol (primeira descri????o na fam??lia). A an??lise quimiom??trica resultou em cinco agrupamentos distintos (C-1 a C-5), onde o C-1 apresentou alta concentra????o de ??- e ??-amirona e aus??ncia de breina e maniladiol; o C-2, com esp??cies potencialmente produtoras de novos triterpenos e de compostos vol??teis; o C-3 com concentra????es m??dias entre as vari??veis analisadas; o C-4 com concentra????o elevada de breina e maniladiol; e o C-5, das esp??cies com alta concentra????o de ??- e ??-amirina. Os ensaios de citotoxicidade demonstraram que dois extratos em hexano (P. cf. rubrum e Trattinnickia peruviana - HrTPE) apresentaram inibi????o contra as linhagens de c??lulas tumorais HCT-8 (93,08 % para ambas as esp??cies) e SF-295 (93,46 e 86,34%, respectivamente). Nos extratos em acetato de etila foram observadas inibi????es por P. giganteum var giganteum e Trattinnickia glaziovii contra as linhagens HCT-8 (88,7 e 92,9%, respectivamente) e MDAM B-435 (97,8 e 96,2%, respectivamente). HrTPE foi submetido a fracionamento, sendo isolado o ??-amiradienol. Do extrato em acetato de etila de P. paniculatum var. modestum foram isolados os triterpenos breina e maniladiol em mistura bin??ria, o ??cido 3-hidroxitirucalla-8,24-dieno-21-??ico puro e em mistura bin??ria com 3-hidroxitirucalla-7,24-dieno-21-??ico. No extrato em acetato de etila de P. bahianum foram isolados os ??cidos urs??lico e olean??lico. Do extrato em acetato de etila de Tetragastris panamensis foram isolados um triterpeno in??dito (??cido 3-oxo- triptoc??lico B) e dois triterpenos descritos pela primeira vez na fam??lia Burseraceae (??cidos triptoc??lico B e 3-oxo-katononico). Uma proposta de bioss??ntese do ??cido 2,3-seco-isobrion??nico a partir do multiflorenol foi apresentada. A quimiossistem??tica da fam??lia foi discutida.

The [M(ESDT)Cl] n (M=Pd or Pt; ESDT=EtO 2 CCH 2 (CH 3 )NCS 2 − , methylamino-acetic acid ethyl ester-dithiocarboxylate) species have been reacted with various amines (py, pyridine; PrNH 2 , n -propylamine; c -BuNH 2 , cyclobutylamine; en, ethylenediamine) in dichloromethane or chloroform with the aim to obtain mixed ligand complexes. The neutral complexes [M(ESDT)(L)Cl] (L=py, PrNH 2 or c -BuNH 2 ) and the ionic species ([M(ESDT)(L) 2 ]Cl and [M(ESDT)(En)]Cl) have been isolated, and characterized by IR and proton NMR spectroscopies. The crystal structure of [Pd(ESDT)(PrNH 2 )Cl] has been determined by X-ray crystallography. The behaviour of the complexes in various solvents was described on the basis of the proton NMR spectra. The complexes and the dithiocarbamato intermediates have been tested for in vitro cytostatic activity against human leukemic HL-60 and HeLa cells.