NRH quinone oxidoreductase 2 (NQO2) is regarded as a mammalian Phase I detoxifying enzyme responsible for reducing quinones to hydroquinones. NQO2 is highly expressed in different types of cancer such as breast and prostate cancer suggesting its participatory role in the progression of these diseases. A potential reason for this is that NQO2 has the ability to modulate the stability of cyclin D1 and activity of NF-ÃŽÂoB and it has been shown that inhibition of NQO2, either genetically or pharmacologically, can alter the pattern of proliferation of cancer cells. However, the biological roles of NQO2 in cancer progression are still ambiguous and need further investigation. A panel of seven ovarian cancer cell lines (OVCs) were screened for the presence and functionality of NQO2. SKOV-3 and TOV-112D cells expressing comparatively the highest and lowest levels of NQO2 were stably transduced to silence and overexpress NQO2 respectively. Pharmacological inhibition was achieved using resveratrol or a series of novel 4-aminoquinolines synthesised in-house. Cell proliferation was monitored by cell counting and clonogenic assays. Flow cytometric analysis was used to determine cell cycle distribution and levels of ROS following modulation of NQO2 function. The expression of cell cycle regulatory markers was determined by Western blot. The contributory roles of NQO2 in determining the cytotoxicity of Adriamycin (ADR) towards OVCs was investigated using MTT assay together with evaluation of P-gp expression and basal ROS levels. In the OVCs panel, NQO2 protein levels and enzymatic activity showed an excellent correlation; with activity varying 36-fold between the cell lines. The sensitivity of OVCs to CB1954 was significantly increased when combined with the NRH-like co-factor, EP0152R. This supports the notion that NQO2 mediates the toxicity of CB1954, which is further confirmed by the strong correlation between cellular NQO2 activity and the responsiveness of the OVC cell lines to CB1954. Hydrazone quinolines showed the highest inhibitiory potency against NQO2 in SKOV-3 when compared to the typical and in-house synthesised quinolines inhibitors. NQO2-overexpressing TOV-112D cells showed more aggressive growth pattern and higher capacity to form colonies than wild-type cells. This was consistently associated with an enhancement in the progression of cells through cell cycle phases and significant reduction in Rb expression. A reduction in ROS levels in NQO2-OE cells may also explain this enhancement in cell growth. Overexpressing NQO2 also resulted in destabilisation of CDK4 and cyclin D1 with significant reduction in their expression levels, and concomitant increase in p-cyclin D1 (Thr286). The involvement of NQO2 in controlling cyclin D1 turnover is also confirmed in SKOV-3 cells when genetic silencing of NQO2 was accompanied by significant reduction in p-cyclin D1 and subsequent stabilisation of cyclin D1 levels. In spite of this, no alterations in the growth pattern of SKOV-3 cells were observed highlighting the impact of cell type on the variations in cellular responses. The role of NQO2 in determining the toxicity of ADR treatment was not proved in OVC cells. This was despite that modulation of NQO2 levels caused significant changes in P-gp expression. The intracellular basal levels of ROS was found to affect the responsiveness of OVCs to ADR as demonstrated when treating SKOV-3 with resveratrol was accompanied by significant increase in ROS levels and concomitant enhancement in the cells’ response to ADR. In conclusion, NQO2 can profoundly alter the proliferation characteristics of OVCs and is a potential therapeutic target for the treatment of this disease. However, the biological functions of NQO2 and its contributory roles in particular pathways are varied among different types of cancer -in other words- are highly dependent on cancer type.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-05-25T14:41:06Z No. of bitstreams: 1 isabeladeoliveirasouza.pdf: 8840752 bytes, checksum: de1f7daf9c0f6025ccc57475de168c79 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-25T14:48:39Z (GMT) No. of bitstreams: 1 isabeladeoliveirasouza.pdf: 8840752 bytes, checksum: de1f7daf9c0f6025ccc57475de168c79 (MD5) / Made available in DSpace on 2017-05-25T14:48:39Z (GMT). No. of bitstreams: 1 isabeladeoliveirasouza.pdf: 8840752 bytes, checksum: de1f7daf9c0f6025ccc57475de168c79 (MD5) Previous issue date: 2017-03-07 / Este trabalho mostra a síntese de 26 derivados aminoquinolínicos e suas avaliações em protozoários. Para uma melhor apresentação do projeto este foi dividido em três séries: a primeira está relacionada à síntese de hidrazonas e a segunda à síntese de bases de Schiff. Os dois núcleos bioativos foram conjugados envolvendo o conceito de hibridação molecular. Já a terceira série envolveu a síntese de complexos de prata, a partir dos ligantes 4-aminoquinolínicos. Todos os compostos foram devidamente caracterizados utilizando técnicas espectroscópicas que permitiram a identificação dos mesmos. Os compostos sintetizados tiveram seu potencial antiparasitário avaliado em parasitos do gênero Leishmania e Plasmodium, que são responsáveis pelas doenças Leishmanioses e Malária, respectivamente. Além disto, tiveram sua toxidez avaliada em células de mamíferos, tendo sido escolhidos macrófagos peritoneais de camundongos. Em relação à atividade leishmanicida, parte dos compostos foram ativos em Leishmania sp, com destaque para o composto 1a em formas amastigotas de L. amazonensis (CI50 de 8,1 µM), resultado próximo ao fármaco utilizado como referência, Miltefosina (CI50 de 4,15µM). A maioria dos compostos não apresentou citotoxicidade expressiva para os macrófagos, com exceção dos compostos 1b, 1f e 2d. Os testes em Plasmodium ainda estão em fase de análise. Os resultados aqui apresentados confirmam o potencial biológico de derivados aminoquinolinas e estimulam a continuidade dos estudos com esta classe de moléculas para o tratamento de doenças parasitárias. / This work demonstrates the synthesis of 26 aminoquinolinic derivatives and their evaluation in protozoa. For a better display, the project was divided into three series: the first is related to hydrazone synthesis and the second to synthesis of Schiff bases. The two bioactive nuclei were conjugated involving the concept of molecular hybridization. The third series involved the synthesis of silver complexes from the 4-aminoquinoline ligands. All components were properly characterized using spectroscopic techniques that allowed their identification. The synthesized compounds had their antiparasitic potential evaluated in parasites of the genus Leishmania and Plasmodium, which are responsible for Leishmaniasis and Malaria, respectively. In addition, their toxicity was evaluated in mammalian cells using murine peritoneal macrophages. Regarding the leishmanicidal activity, part of the compounds presented activity against Leishmania sp, with emphasis on compound 1a in amastigotes of L. amazonensis (IC 50 of 8.1 μM), a close result to the reference drug Miltefosine (IC 50 of 4.15 μM). Most compounds did not present expressive cytotoxicity for macrophages, except for compounds 1b, 1f and 2d. Plasmodium tests are still under review. These results confirm the potential biological of aminoquinoline derivatives and stimulate further studies with this class of molecules for the treatment of parasitic diseases.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2016-08-08T14:06:04Z No. of bitstreams: 1 arturenemarialinocarmo.pdf: 9897809 bytes, checksum: 80040aff8b9e00693411113ba82ed918 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-08-08T14:10:07Z (GMT) No. of bitstreams: 1 arturenemarialinocarmo.pdf: 9897809 bytes, checksum: 80040aff8b9e00693411113ba82ed918 (MD5) / Made available in DSpace on 2016-08-08T14:10:07Z (GMT). No. of bitstreams: 1 arturenemarialinocarmo.pdf: 9897809 bytes, checksum: 80040aff8b9e00693411113ba82ed918 (MD5) Previous issue date: 2013-05-24 / A tese de doutorado intitulada Síntese de Derivados Aminoquinolínicos e Nucleotídeos Não-Naturais, Potenciais Agentes Antiparasitários e Antibacterianos está apresentada em dois capítulos que descrevem a síntese de substâncias com potencial atividade antiparasitária e antibacteriana. Foram obtidos 46 compostos nesse trabalho, 31 compostos do capítulo 1, sendo 15 inéditos, e o restante foi desenvolvido durante a realização do capítulo 2, sendo 8 inéditos. O capítulo 1 apresenta a síntese de derivados quinolínicos funcionalizados usando a reação do tipo “click” para obtenção dos conjugados aminoquinolina/ácido cólico. Inicialmente é apresentada a obtenção dos derivados aminoquinolinas, aminoalcinoquinolinas e aminodialcinoquinolinas, estes dois últimos foram usados na reação do tipo “click” para obtenção dos derivados triazólicos, alguns também foram utilizados como ligantes na síntese de complexos de Pt (II). Um estudo da relação estrutura/atividade dos compostos sintetizados foi realizado, permitindo a verificação da importância biológica dos compostos. O segundo capítulo envolveu a síntese de derivados nucleotídicos não- naturais em que o grupo fosfato é substituído por um triazol e foi realizado no Instituto de Química Molecular de Reims na Universidade de Reims Champagne-Ardenne CNRS sob a coordenação da professora Dra. Pascale Clivio, / The doctoral thesis entitled “Synthesis of Derivatives Aminoquinolínicos and Non-Natural Nucleotides, Potential Antiparasitic and Antibacterial Agents” is presented in two chapters that describe the synthesis of substances with potential antibacterial and antiparasitic activity. 46 compounds were obtained in this work, Chapter 1 31 compounds, including 16 unpublished, and the remainder was developed during the course of Chapter 2, and 8 unpublished. Chapter 1 presents the synthesis of functionalized quinoline using the click reaction to obtain the conjugated aminoquinoline/cholic acid. Initially it is presented to obtain the derivatives aminokynolines, and aminoalkynoquinolines aminodialkynoquinolines, these two latter were functionalized using reaction type click to prepare the triazoles, some have also been used as binders in the synthesis of platinum (II) complexes. A study of the structure/activity of the synthesized compounds was performed by allowing verification of the biological importance of the compounds. The second chapter involved the synthesis of non-natural nucleotide in which the phosphate group is replaced by a triazole and was conducted at the Institute of Molecular Chemistry of the University of Reims Champagne-Ardenne Reims CNRS under the supervision of Professor Dr. Pascale Clivio,

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-05-02T11:30:44Z No. of bitstreams: 1 robertareissoares.pdf: 1821120 bytes, checksum: 37f4636d8bd6ef3623072173760d6105 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-12T15:51:49Z (GMT) No. of bitstreams: 1 robertareissoares.pdf: 1821120 bytes, checksum: 37f4636d8bd6ef3623072173760d6105 (MD5) / Made available in DSpace on 2017-05-12T15:51:49Z (GMT). No. of bitstreams: 1 robertareissoares.pdf: 1821120 bytes, checksum: 37f4636d8bd6ef3623072173760d6105 (MD5) Previous issue date: 2013-02-20 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / A malária permanece ao longo dos anos como emergência global em saúde pública. São aproximadamente 87 milhões de casos e 106.820 mortes ocorridas anualmente no mundo. As principais estratégias atuais no controle da doença se baseiam no diagnóstico precoce e tratamento oportuno dos casos, devendo ser considerada a resistência dos parasitos a praticamente todos os fármacos atualmente em uso, o que torna urgente a busca por novos antimaláricos. Neste contexto, análogos de 4-aminoquinolinas e 6-mercaptopurinas contendo 1,2,3-triazol ou esteroide foram sintetizados e tiveram sua atividade antimalárica investigada utilizando o teste supressivo de Peters em modelo murino de infecção por Plasmodium berghei NK65. Dentre os 11 análogos avaliados, 5 exibiram atividade antimalárica significativa, alcançando percentuais de supressão de até 81% em relação ao controle não tratado. Além da atividade antimalárica, um composto promissor não pode apresentar efeitos indesejáveis às células do hospedeiro, sendo assim foram avaliadas a citotoxicidade e atividade hemolítica dos análogos. A maioria dos compostos foram considerados seguros às células do hospedeiro mesmo nas maiores concentrações avaliadas. Portanto, estes análogos merecem ser objeto de futuras investigações visando compor novos antimaláricos. / Malaria remains over the years as a global public health emergency. There are approximately 87 million cases and 106.820 deaths worldwide each year. The main current strategies to control the disease are based on early diagnosis and appropriate treatment cases and should be considered the resistance of parasites to almost all drugs currently in use, which makes urgent the search for new antimalarials. In this context, analogs of 4-aminoquinolines and 6-mercaptopurines containing 1,2,3-triazole or steroid were synthesized and investigated for their antimalarial activity using the 4-day suppressive test described by Peters in a murine model of infection by Plasmodium berghei NK65. Among the 11 analogues evaluated, 5 exhibited significant antimalarial activity, reaching percentages of suppression up to 81% compared to untreated control. Besides antimalarial activity, a promising compound cannot have undesirable effects on host cells, thus was evaluated the cytotoxicity and hemolytic activity of analogs. Most compounds were considered safe to host cells even at the highest concentrations evaluated. So, this analogs deserve to be object of future investigations aiming compose new antimalarials.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-08-17T11:16:52Z No. of bitstreams: 1 lucianamariaribeiroantinarelli.pdf: 16756474 bytes, checksum: 2304892df326fd4997c78bc7f0870bae (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-17T11:34:54Z (GMT) No. of bitstreams: 1 lucianamariaribeiroantinarelli.pdf: 16756474 bytes, checksum: 2304892df326fd4997c78bc7f0870bae (MD5) / Made available in DSpace on 2017-08-17T11:34:54Z (GMT). No. of bitstreams: 1 lucianamariaribeiroantinarelli.pdf: 16756474 bytes, checksum: 2304892df326fd4997c78bc7f0870bae (MD5) Previous issue date: 2017-02-15 / As leishmanioses representam um grande problema de saúde pública mundial com sérias limitações na quimioterapia atual, como: número limitado de fármacos, baixa eficácia, custo elevado, crescente resistência parasitária e toxicidade. O objetivo do presente trabalho foi avaliar a atividade de uma série de dez derivados de 4- aminoquinolinas em espécies de Leishmania relacionadas a leishmaniose tegumentar. Avaliar os possíveis mecanismos de ação do composto com atividade antileishmanial promissora, identificando as organelas alvo e os processos de morte celular desencadeados no parasito, bem como a sua eficácia in vivo. Dentre os compostos avaliados, o derivado AMQ-j foi o mais ativo, com atividade expressiva em L. amazonensis e L. braziliensis (CI50 menor que 6.0 μg/mL e 2,5 μg/mL em promastigotas e amastigotas intracelulares, respectivamente) para ambas as espécies avaliadas. O composto AMQ-j apresentou baixa toxicidade para macrófagos murinos (CC50> 40.0 μg/mL), sendo mais tóxico para amastigotas intracelulares (Índice de Seletividade>12,0). Os resultados preliminares acerca do modo de ação apontam que o composto AMQ-j induziu drásticos efeitos na mitocôndria do parasito e caracterizado pelo colapso do potencial de membrana mitocondrial (ΔΨm), inchaço da organela, aumento na produção de Espécies Reativas do Oxigênio (EROS) e acúmulo de corpúsculos lipídicos (CLs) no citoplasma. O tratamento com AMQ-j induziu nas formas promastigotas uma série de alterações bioquímicas e celulares sugestivas de morte por apoptose-like como redução do volume celular, exposição de fosfatidilserina no folheto externo da membrana plasmática, manutenção da integridade da membrana plasmática e alterações drásticas no núcleo celular evidenciadas por meio da desorganização da cromatina e fragmentação do DNA. O efeito do composto em promastigotas está também associado à indução de morte por autofagia evidenciada pelo aumento de vacúolos autofágicos, presença de corpos multivesiculares dentro de vacúolos, vesículas citoplasmáticas e acúmulo de compartimentos acídicos no citoplasma dos promastigotas. O composto também induziu fragmentação do DNA dos amastigotas intracelulares de modo seletivo, sem induzir fragmentação da célula hospedeira. Estudos in silico sugerem que AMQ-j é um potencial inibidor da tripanotiona redutase (TryR), enzima fundamental na defesa antioxidante do parasito. Estudos in vivo em modelo murino de infecção com L. amazonensis demonstraram a eficácia do AMQ-j pela via intralesional na redução do tamanho da lesão e da carga parasitária, sem indução de toxicidade hepática, cardíaca e renal. Os estudos de predição in silico relacionados a propriedades farmacocinéticas (ADMET) e características físico-químicas (regra de Lipinsky) sugerem que AMQ-j pode ser utilizado pela via oral. O efeito leishmanicida do composto está associado a múltiplos alvos, desencadeando a morte do parasito por diferentes vias, como apoptose e autofagia. O efeito in vivo do composto aponta para a necessidade da continuidade dos estudos no intuito de melhor estabelecer o seu efeito leishmanicida. / Leishmaniasis represents a major global public health problem with serious limitations in current chemotherapy, such as limited number of drugs, low efficacy, high cost, increasing parasitic resistance and toxicity. The objective of the present study was to evaluate a series of ten 4-aminoquinolines derivatives (AMQs) on Leishmania species related to tegumentary leishmaniasis. It was also evaluated the possible mechanisms of action of a compound with promising leishmanicidal activity, identifying the target organelles and the type of death triggered in the parasite, as well as to its leishmanicidal effect in vivo. Among the evaluated compounds, the AMQ-j derivative was the most active, with expressive activity on L. amazonensis and L. braziliensis (IC50 less than 6.0 μg/mL and 2.5 μg/mL against intracellular promastigotes and amastigotes, respectively) for both evaluated species. Furthermore, AMQ-j showed low toxicity for murine macrophages (CC50> 40.0 μg/mL) being more destructive to the intracellular parasites (selectivity index > 12.0). Preliminary studies about the mode of action showed that AMQ-j compound induced marked effects on the parasite mitochondria, characterized by mitochondrial membrane potential collapse (ΔΨm), organelle swelling, increased of Reactive Oxygen Species (ROS) production and lipidic bodies accumulation in the cytoplasm. The treatment with AMQ-j induced in the promastigote forms a series of biochemical and cellular alterations which suggest apoptosis-like death, including reduction of cellular volume, phosphatidylserine exposure on the outer leaflet of the plasma membrane, maintenance of plasma membrane integrity and drastic changes in the cell nucleus evidenced by chromatin disorganization and DNA fragmentation. The effect of AMQ-j in promastigote forms is also associated with the induction of death by autophagy evidenced by the increase of autophagic vacuoles, presence of multivesicular bodies inside vacuoles, cytoplasmic vesicles and accumulation of acidic compartments in the promastigote cytoplasm. The compound also induced DNA fragmentation of intracellular amastigotes selectively, without inducing host cell fragmentation. In silico studies suggest that this compound is a potential inhibitor of key redox enzyme trypanothione reductase (TryR). In vivo studies in murine infection model of L. amazonensis demonstrated the efficacy of AMQ-j by the intralesional route, reducing lesion size and parasite load, without induction of hepatic, cardiac and renal toxicity. The in silico prediction studies on pharmacokinetic properties (ADMET) and physico-chemical characteristics (Lipinsky's rule) suggest that AMQ-j can be used orally. Taken together, the results suggest that the leishmanicidal effect of the compound is associated with multiple targets and triggers parasite death through different pathways, including apoptosis and autophagy. The in vivo effect indicates that studies with this compound should be continued in order to better establish its leishmanicidal effect.

In this work, the evaluation of the antinociceptive and anti-inflammatory activities of two series of rationally designed derivative was performed. The first series belong to the class hydrazones thiophene derivative is formed by nine numbered 5a-i. The second is termed aminoquinolínicos BAQ, 7-Cl-MAQ, 2-CF3-MAQ e 7-CF3-MAQ, synthesized from structural modifications in the molecule chloroquine. The two series were synthesized and subjected to testing for antinociceptive activity (writhing test induced by acetic acid and testing of formalin-induced nociception), testing for acute anti-inflammatory activity (zymosan A/carrageenan-induced peritonitis) and chronic (experimental arthritis syndrome induced by Freund's complete adjuvant). The first series was still subjected to a theoretical study in silico toxicity Osiris® program through which the data showed no significant toxic effects. Forthe study, Swissmale mice (20-30g) and Wistar rats (130-170g) rats line ages were used. The drugs dexamethasone, dipyrone and indomethacin were used as reference standards for the two series. All substances were administered 40 minutes before stimulation. The number of hydrazones thiophene derivatives was administered orally, while aminoquinolines were administered intraperitoneally. All derivatives of the first series exhibited anti-inflammatory activity and high potency and antinociceptive efficacy highlighting derivatives 5a and 5d were submitted to chronic testing of arthritis syndrome, being able to reduce the paw edema from the 4th day of treatment without producing renal, hepatic or gastric toxicity under the conditions tested. The derived series of aminoquinolines induced anti-inflammatory and antinociceptive activity, being selected for the chronictest the BAQ derivatives and 7-CF3-MAQ. The treatment for seven days this series showed a significant reduction in the size of the legs of animals, especially from the 4th day of treatment, without changing the levels of liver enzymes, change the weight or generate gastric toxicity. These results demonstrate that administration of derivatives of thiophene and hydrazones series aminoquinolines systemically generated antinociceptive and anti-inflammatory activity, but further studies are needed to elucidate their mechanisms of action. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Neste trabalho foi realizada a avaliação das atividades antinociceptiva e anti-inflamatória de duas séries de derivados racionalmente planejados. A primeira série pertence à classe hidrazonas tiofeno, é formada por nove derivados numerados de 5a-i.A segunda são aminoquinolínicos denominadas de BAQ, 7-Cl-MAQ, 2-CF3-MAQe 7-CF3-MAQ, sintetizadas a partir de modificações estruturais na molécula de cloroquina. As duas séries foram sintetizadas e submetidas a ensaios para atividade antinociceptiva (ensaio de contorções abdominais induzidas por ácido acético e ensaio de nocicepção induzida por formalina), ensaios para atividade anti-inflamatória aguda (peritonite induzida por carragenina / zymosan A) e crônica (síndrome experimental da artrite induzida por adjuvante completo de Freund’s). A primeira série foi ainda submetida a um estudo in silico de toxicidade teórica através do programa Osiris® o qual os dados não demonstraram efeitos tóxicos significativos. Para a realização do estudo foram utilizados camundongos machos da linhagem Swiss (20-30g) e ratos Wistar (130-170g). Os fármacos dexametasona, dipirona e indometacina foram utilizados como padrões de referência para as duas séries. Todas as substâncias foram administradas 40 minutos antes do estímulo. A série dos derivados hidrazonas tiofeno foi administrada por via oral, enquanto que aminoquinolinas foram administradas por via intraperitoneal. Todos os derivados da primeira série exibiram atividade anti-inflamatória e elevada potência e eficácia antinociceptiva destacando-se os derivados 5a e 5d que foram submetidos ao ensaio crônico de síndrome de artrite, sendo capazes de reduzir o edema de pata a partir do 4º dia de tratamento, sem produzir efeitos tóxicos renais, hepáticos ou gástricos nas condições testadas. Os derivados da série das aminoquinolinas induziram atividade anti-inflamatória e antinociceptiva, sendo selecionados para o ensaio crônico os derivados BAQ e 7-CF3-MAQ. O tratamento por sete dias dessa série evidenciou uma significativa redução do tamanho das patas dos animais, especialmente a partir do 4º dia de tratamento, sem alterar os níveis das enzimas hepáticas, alterar o peso ou gerar toxicidade gástrica. Esses resultados demonstram que a administração dos derivados das séries hidrazonas tiofeno e aminoquinolinas por via sistêmica gerou atividade antinociceptiva e anti-inflamatória, no entanto novos estudos são necessários para elucidação dos seus mecanismos de ação.