Yes / Electron‐deficient half‐sandwich complexes are a class of under‐studied organometallics with demonstrated potential as metallodrug candidates. The present study investigates the effect of two 16‐electron organoruthenium complexes ([( p‐ cym)Ru(benzene‐1,2‐dithiolato)] ( 1 ) and [( p ‐cym)Ru(maleonitriledithiolate)] ( 2 )) on the cell viability of non‐immortalised human lymphocytes from healthy individuals. The genotoxic effects of 1 and 2 in lymphocytes using the Comet and cytokinesis‐block micronucleus assays is also investigated. Gene expression studies were carried out on a panel of genes involved in apoptosis and DNA damage repair response. Results show that the two 16‐electron complexes do not have significant effect on the cell viability of human lymphocytes from healthy individuals. However, an increase in DNA damage is induced by both compounds, presumably through oxidative stress production. / This project was supported by the Royal Society (University Research Fellowship No. UF150295 to NPEB), the University of Bradford (RDF Award), and by the Academy of Medical Sciences/the Wellcome Trust/ the Government Department of Business, Energy and Industrial Strategy/ the British Heart Foundation Springboard Award [SBF003\1170 to NPEB].

The thesis “Ancillary Ligand Effects on the Anticancer Activity of Ruthenium (II) Piano Stool Complexes” is an effort to design better antitumor metallodrugs based on ruthenium(II) complexes with various H-bond donor/acceptor ligands and to understand their mechanism of action. Chapter 1 presents a brief review of metallodrugs and their mechanism of action. Different classes of metallodrugs are discussed. A short discussion on ruthenium based anticancer drugs and their established mechanism of action is also included in this chapter. Chapter 2 deals with the synthesis, characterization and anticancer activity of Ru(II) complexes with P(III) and P(V) ligands. The effect of a strong hydrogen bond acceptor on the cytotoxicity of the complexes has been investigated which allows comparison of complexes with ligands possessing a strong hydrogen bond donor or hydrogen bond acceptor. Partial oxidation of the tertiary phosphine ligands leads to a decrease in cytotoxicity of the ligand, while coordination to ruthenium resulted in a significant increase in the cytotoxicity. A molecular mechanism of action for these complexes was suggested on the basis of various biophysical studies. These complexes bind DNA through non-intercalative interactions which lead to the destabilization of the double helix of the DNA and also unwinding of the negatively supercoiled DNA. Results show that the presence of a hydrogen bond acceptor on the ligand is not capable of enhancing interactions with DNA in comparison with hydrogen bond donor groups. Cellular studies of these complexes showed that inhibition of DNA synthesis and apoptosis occur on treatment with these complexes. Interestingly, these complexes are found to be not only cytotoxic but also antimetastatic. Chapter 3 deals with the synthesis, characterization and anticancer activity of Ru(II) complexes with biologically active S containing heterocyclic ligands and their mechanistic study. Complexation of ruthenium with mercaptobenzothiazole (MBT) gave the most cytotoxic complex (H3) in the series. Heterocyclic Ru(II) complexes behave differently as evidenced by cellular and biophysical studies. Unlike phosphine complexes, H3 shows biphasic melting of DNA at higher concentrations which suggests two different types of interaction with DNA. Chapter 4 deals with synthesis and characterization of water soluble multiruthenated hydrophilic ruthenium(II) complexes with urotropine. An increase in cytotoxicity and binding affinity has been observed with increase in the number of ruthenium atoms per molecule. The complex with three ruthenium atoms showed the best activity. However cytotoxicity of the complexes decreases with decrease in the lipophilicity of the complexes. Chapter 5 describes studies on the interaction of Ru complexes with water, ss-DNA, AMP, GMP and GSH by various spectroscopic techniques. Hydrolysis of Ru-Cl bond in the complexes correlates with the cytotoxicity. Chapter 6 reports the summary of the observations of the thesis and the future prospects of metallodrugs.

No description available.

The application of ICP-MS to the fields of proteomics and genomics has arisen in part due to its ability to detect and quantify trace levels of S and P, which are major constituents in proteins and nucleic acids respectively. The development of collision/reaction cell technology and high resolution instruments has enabled these biologically important elements to be measured and quantified at the pg - ng ml-1 level. Despite these advances, the detection limits of P and S are still inferior compared to other elements. Oligonucleotides containing biotin functionality were labelled with Au nano-particles attached to a streptavidin protein to achieve site specific labelling, with 100% labelling efficiency. Each nano-particle contained ~86 Au atoms, resulting in an 882 fold signal enhancement for 24 base length oligonucleotides. However, this enhancement factor was only observed when one oligonucleotide bound to one nano-particle in a 1:1 ratio. Much lower Au labelling efficiencies and signal enhancements were observed when thiolated oligonucleotides were labelled with maleimide functionalised gold nano-particles. This was attributed to the extensive and difficult sample preparation steps that were required prior to labelling. The detection and quantification of adducts formed between DNA and the Pt anti-cancer drugs cisplatin and oxaliplatin were also investigated with ICP-MS. Acid digestion of the carbon based DNA matrix enabled Pt adducts to be quantified at low dose rates of 1 Pt atom per 1 500 000 nucleotides in ~12 μg DNA. Such sensitive mass spectrometric determinations could be employed in clinical tests to detect and quantify low level adducts formed in patients in-vivo. To complement ICP-MS analysis, electrospray ionisation linear ion trap mass spectrometry was employed to study the interaction of oxaliplatin with the four DNA nucleobases. Multiple stage mass spectrometry enabled detailed Pt-nucleobase adduct fragmentation pathways to be established. The method of DNA detection using P in conjunction with the collision cell, or cool plasma to form PO+ was also demonstrated and the limitations of the method, namely, polyatomic interferences and severe matrix effects were highlighted.

Os fármacos antiinflamatórios não-esteróides (FAINEs) são amplamente utilizados no combate a processos inflamatórios e dores, mas apresentam restrição de uso em razão de sérios efeitos colaterais sobre o trato gastrointestinal. A atividade biológica de complexos metálicos tem sido objeto de pesquisa de grande interesse na área de metalofármacos e compostos do tipo Cu(II)-FAINEs apresentam boa atividade antiinflamatória e efeitos colaterais reduzidos em relação aos fármacos orgânicos. Nesse trabalho, com o objetivo de contribuir para ampliar os estudos sobre o desenvolvimento de alternativas aos FAINEs, foram preparados e caracterizados complexos cobre(lI) com ibuprofeno, indometacina, naproxeno, sulindaco, meloxicam. Os compostos foram imobilizados em Hidrotalcita, um hidróxido duplo lamelar (HDL) de magnésio e alumínio (Mg/AI = 3) que é biocompatível e tem uso como antiácido estomacal. As interações dos metalofármacos com o HDL geraram materiais híbridos bioinorgânicos do tipo Cu-FAINElHDL, nos quais os complexos podem estar presentes de duas maneiras: intercalados entre as Ia meias ou adsorvidos nas superfícies externas do hidróxido duplo lamelar, dependendo do solvente utilizado. A intercalação de complexos neutros é favorecida em solvente misto álcooVamida no qual a Hidrotalcita sofre esfoliação promovida por um processo de hidrólise da amida. A estabilidade alguns dos complexos e dos materiais híbridos em condições gástricas simuladas (pH e temperatura) e as atividades antiinflamatória, analgésica e ulcerogênica in vivo dos sistemas contendo indometacina (Indo) foram investigadas. A intercalação do complexo Cu-Indo no HDL favorece sua estabilização, contribuindo para potencializar a sua atividade farmacolágica. Os materiais híbridos bioinorgânicos obtidos neste trabalho apresentam propriedades interessantes com vistas a potencial aplicação como sistemas de liberação controlada de fármacos. / Non-steroidal anti-inflammatory drugs are widely consumed to treat inflammatory diseases and pain but their clinical use are limited due to serious side-effects on the gastrointestinal tract. The bioactivity of metal complexes exhibits great interest in metal-based drug research. Cu(II)-NSAID compounds show good anti-inflammatory property and decreased side-effects compared to their organic parent drugs. This work aimed to contribute for development of alternative NSAIDs. Cu-NSAlD compounds containing the drugs Ibuprofen, Indomethacin, Naproxen, Sulindac and Meloxicam were synthesized and characterized. The compounds were also immobilized on Hydrotalcite, a layered double hydroxide (LDH) of magnesium and aluminum (Mg/Al = 3), that is biocompatible and used as stomach antacid. The interactions of the copper drugs with LDH led to Cu-NSAID/LDH- bioinorganic hybrid materiais. Two kinds of complex-LDH interactions were observed by changing the solvent: intercalation between LDH layers or adsorption on the LDH external surfaces. The intercalation of neutral complexes is favored in an alcohollamide solvent mixture where Hydrotalcite undergoes exfoliation process promoted by the amide hydrolysis. The stability of some complexes and their correspondent hybrid materials under gastric conditions (pH and temperature) and also in vivo anti-inflammatory, analgesic and ulcerogenic activities for Indomethacin (Indo)-containing systems were investigated. The stabilization of the Cu-Indo structure is promoted by intercalation of the complex into the LDH layers, contributing to increase its pharmacological activity. The bioinorganic hybrid materials here investigated also exhibit interesting properties for applications as controlled drug delivery systems.

Metalofármacos baseados em rutênio têm se mostrado promissores com relação à atividade anticancerígena frente a diversos tipos de tumores. Nosso grupo de pesquisa dedica-se ao estudo de compostos contendo o centro dimetálico de valência mista Ru2(II,III) coordenado a ligantes derivados de Faines (Fármacos anti-inflamatórios não esteroides), tendo demostrando o potencial desses complexos frente a glioma. O entendimento do modo de ação destes complexos requer o estudo de suas interações com biomoléculas presentes no meio biológico. Neste cenário, o presente trabalho teve por objetivo investigar a interação de três complexos de dirutênio-Faines, ou RuFaines, [Ru2(ibp)4Cl], [Ru2(ceto)4Cl] e [Ru2(npx)4(H2O)2]PF6 (ibp = ibuprofenato, ceto = cetoprofenato e npx = naproxenato), e também do precursor [Ru2(O2CCH3)4Cl], RuAc, com as principais proteínas presentes no soro humano, transferrina e albumina. Os complexos foram sintetizados e caracterizados conforme metodologias desenvolvidas no grupo. A interação destes complexos com a transferrina, em suas formas apo e holo, e com a albumina foi avaliada por técnicas como espectroscopia eletrônica, dicroísmo circular, fluorescência, e realizaram-se estudos de ultrafiltração com análise do aduto formado por ICP-OES e espectrometria de massas. Além disso, fez-se um estudo de captação celular dos complexos RuFaines por células de glioma humano da linhagem U-87. Os resultados demonstraram que os complexos de dirutênio-Faines interagem com ambas as proteínas séricas (transferrina (apo e holo) e albumina), de modo semelhante, mas que é distinto daquele observado para o complexo RuAc. A presença de íons Fe(III) nos sítios específicos da transferrina não afetou a interação dos complexos RuFaines, enquanto que um comportamento diferente foi observado para o RuAc. Verificou-se que todas as proteínas avaliadas (albumina, apo-transferrina e holo-transferrina) apresentam capacidades similares de retenção dos complexos (~ 70% da quantidade de Ru adicionada inicialmente), independentemente da natureza do ligante carboxilato coordenado. Estudos de captação celular mostraram que a interação dos complexos RuFaines com a transferrina não contribuiu para modificar a capacidade de entrada desses complexos na célula, em comparação com os metalofármacos livres. Em alguns casos, inclusive, a formação de aduto com a apo-transferrina teve um efeito contrário, diminuindo a captação de rutênio. Dessa forma, concluiu-se que o ciclo da transferrina provavelmente não é a principal rota de entrada nas células para os complexos estudados. / Ruthenium metallodrugs have shown promising antitumor activity against to several tumor types. Our research group is dedicated to study compounds containing the mixed-valence Ru2(II,III) dimetallic center coordinated to NSAIDs (nonsteroidal anti-inflammatory drugs) derived ligands, and have demonstrated the potential of these complexes against glioma. The understanding of the mode of action of these complexes requires the study of their interactions with biomolecules present in biological environment. In this scenario, the present work aimed to investigate the interaction of three complexes of diruthenium-NSAIDs, or RuNSAIDs, [Ru2(ibp)4Cl], [Ru2(ceto)4Cl] and [Ru2(npx)4(H2O)2]PF6 (ibp = ibuprofenate, ceto = ketoprofenate, npx = naproxenate), and also of the precursor [Ru2(O2CCH3)4Cl], RuAc, with the major proteins present in the human serum, transferrin and albumin. The complexes were synthesized and characterized according to methods developed in our group. The interaction of these complexes with transferrin, in the two forms apo and holo, and with albumin was evaluated by techniques as electronic spectroscopy, circular dichroism, fluorescence, and ultrafiltration studies accompanied by analysis of adducts by ICP-OES and mass spectrometry. Moreover, cellular uptake studies of the RuNSAIDs complexes by U-87 human glioma cells line were performed. The results demonstrated that the diruthenium-NSAIDs complexes interact with both proteins (transferrin (apo and holo) and albumin), in a similar way, but that is distinct of that observed for the RuAc complex. The presence of Fe(III) ions in transferrin specific binding sites did not affect the interaction of the RuNSAID complexes with the protein, while a different behavior was shown by RuAc. All the proteins studied here (albumin, apo-transferrin and holo-transferrin) showed similar capabilities for retention of the complexes (~ 70 % of the initial amount of Ru added), independently of the nature of the coordinated carboxylate ligand. Cellular uptake studies showed that the interaction of the RuNSAIDs complexes with transferrin did not contribute to modify the internalization capacity of these complexes, in comparison with the free metallodrugs. In some cases, the adduct formation with apo-transferrin showed an opposite effect, leading to the decrease of ruthenium uptake. The findings led to the conclusion that transferrin cycle probably is not the main entry way to the cells for the studied complexes.

A bioatividade de complexos metálicos é de grande interesse atual na área de pesquisa de novos medicamentos. Fármacos anti-inflamatórios não-esteróides (FAINEs) são amplamente consumidos para o tratamento de doenças inflamatórias e de dor, mas seu contínuo uso pode ocasionar sérios efeitos colaterais ao trato gastrointestinal (TGI). Metalofármacos alternativos de Cu-FAINEs causam menores danos ao TGI, sem perder a atividade anti-inflamatória. No entanto, alguns destes são pouco solúveis. Adicionalmente, o desenvolvimento de materiais híbridos de FAINEs/polímeros para liberação modificada dos fármacos é interessante do ponto de vista de se aumentar a eficácia terapêutica e minimizar problemas de toxicidade. O objetivo deste trabalho foi investigar metodologia, com emprego de secagem via spray-dryer, para a preparação de materiais híbridos contendo complexos de cobre(II) com os fármacos naproxeno e sulindaco incorporados em acetato de celulose. Dois tipos de materiais híbridos foram obtidos para cada metalofármaco: CuFAINE/AC e CuFAINE/AC/OM, sendo o segundo preparado em presença de óleo mineral (OM). A caracterização destes produtos foi efetuada por análises elementares, espectroscopia vibracional no infravermelho, difratometria de raios X, análise termogravimétrica acoplada a espectrômetro de massas, ressonância paramagnética eletrônica, microscopia eletrônica de varredura e espalhamento de luz dinâmico. Os métodos utilizados levaram à interação dos metalofármacos de CuFAINEs com a matriz polimérica biocompatível de acetato de celulose, gerando materiais híbridos bioinorgânicos-orgânicos. Duas espécies de CuFAINEs: uma dimérica mantendo a estrutura original ([Cu2(FAINE)4(Laxial)2]) e outra monomérica - provavelmente do tipo ([Cu(FAINE)2(Laxial)2]) - foram identificadas nestes materiais. As partículas apresentaram tamanho nanométrico. Os resultados aqui obtidos abrem novas perspectivas para futuros estudos que visem à investigação da viabilidade de usar sistemas como estes para liberação mofidicada dos metalofármacos de CuFAINEs / The bioactivity of metal complexes has current and high interest in the field of development of new drugs. Non-steroidal anti-inflammatory drugs (NSAIDs) are widely consumed to treat inflammatory diseases and pain. The continuous use of these drugs causes serious side-effects on the gastrointestinal tract (GT). Alternative copper(II)-metallodrugs (CuNSAIDs) show lower GT side-effects without loss of anti-inflammatory activity. However, some of them are poorly water-soluble. In addition, the development of NSAID/biocompatible polymeric hybrid materials for modified-release of pharmaceuticals is of interest to increase therapeutic efficacy and to lower toxicity of drugs. The aim of this work was to investigate a methodology adding the use of spray-drying technique to prepare hybrid materials containing copper(II) complexes with naproxen and sulindac drugs incorporated into cellulose acetate (CA). Two different materials were obtained for each metallodrug: CuNSAID/CA and CuNSAID/CA/MO, being the second one prepared in the presence of mineral oil (MO). The characterization of these products was conducted by elemental analysis, vibrational spectroscopy, powder x-ray diffraction, thermogravimetric analysis coupled to mass spectrometry, electron paramagnetic resonance, scanning electron microscopy and dynamic light scattering. The developed methodologies led to the interaction between the CuNSAIDs and the polymeric matrix generating bioinorganic-organic hybrid materials. Two CuNSAIDs species: a dimeric one that kept the complex original structure [Cu2(FAINE)4(Laxial)2] and a monomeric species, ([Cu(FAINE)2(Laxial)2] were identified in these materials. The particles have shown nanometric size. The obtained results open up new perspectives for future studies with the aim of investigating the availability of using this type of system for modified release of CuNSAIDs metallodrugs.

Neste trabalho sintetizaram-se novos complexos imínicos e diimínicos de Cu(II) derivados da isatina, um indol endógeno, que foram então extensivamente caracterizados por análise elementar, medidas de condutividade molar, técnicas espectroscópicas: IV, UV/Vis e EPR, e por espectrometria de massa, ESI-MS. Estes compostos apresentaram equilíbrio ceto-enólico em solução, variando a geometria ao redor do íon Cu(II), ao se alterar o pH do meio, indo de tetraédrico em meio ácido à tetragonal em meio básico. Para a elucidação do papel do cobre no mecanismo de oxidação de carboidratos pelo oxigênio molecular, realizaram-se estudos cinéticos tendo como catalisadores estes complexos de cobre. Determinou-se a uma lei cinética global mais abrangente para estas reações, incluindo uma etapa dependente do cobre (a concentrações bem baixas), seguida de outra independente do metal. As etapas no mecanismo proposto, sob condições de pseudo-primeira ordem, combinam transferência eletrônica intramolecular, com provável redução do íon de cobre pelo substrato, levando a espécies muito reativas (OH•-, O2•-, H2O2, CO2•-), responsáveis pelo processo de iniciação e propagação, e a formação de produtos carbonílicos de cadeia curta (glicolato, glicerato e formiato). Para o estudo da interação metal-carboidrato, importante para se entender melhor o papel do cobre frente a este ligante biológico, complexos de Cu(II) com monossacarídeos foram sintetizados e caracterizados por análise elementar e termogravimétrica, medidas de condutividade molar, espectroscopias UV/Vis, IV e EPR, além da espectroscopia Raman, através da qual investigou-se o modo de ligação do carboidrato ao metal em cada um destes complexos. Foram ainda preparados e caracterizados por análise elementar, medidas de condutividade molar, espectroscopias UV/Vis, IV, EPR, ESI-MS e CD dois novos complexos imínicos quirais de Cu(II), com ligantes do tipo aminocarboidrato, e que também foram utilizados para estudos biológicos. Estudos de atividade biológica foram feitos in vitro, usando as linhagens celulares tumorais promonocítica sanguínea U937 e neuroblastoma SH-SY5Y, com os complexos imínicos de cobre, principalmente aqueles derivados da isatina. As células tratadas com os complexos mais ativos sofreram apoptose, verificada por ensaios citofluorimétricos, em que os complexos agiram em diferentes fases do ciclo celular de cada linhagem (fase G1, S ou G2/M). Através da técnica citofluorimétrica foi observada também a geração de radicais livres na célula e, através de ensaios imunológicos, determinou-se a quantidade de proteínas citoplasmáticas carboniladas e glicosiladas, geradas após os tratamentos com os compostos, em diferentes tempos de incubação. De uma maneira geral, estes estudos indicaram modulação da atividade biológica, com um comportamento antiproliferativo muito diferente, indo de baixa eficácia até alta eficiência. Dentre os compostos mais ativos, pode ser observada uma especificidade diferente, tanto com relação ao tipo de célula quanto ao seu modo de ação, evidenciando sua potencial aplicação como agentes antitumorais. / In this work, novel imine and diimine copper(II) complexes with ligands derived from isatin, an endogenous indol, were synthesized, and extensively characterized by elemental analysis, conductivity measurements, spectroscopic techniques (IR, UV/Vis, and EPR), and electrospray mass spectrometry (ESI-MS/MS). These compounds showed a keto-enolic equilibrium in solution, with variations in the geometry around the copper ion with increasing pH, varying from a more tetrahedral configuration in acidic medium to a tetragonal one in basic solution. In order to elucidate the role of copper in the carbohydrate oxidation by molecular oxygen, kinetic studies were performed using these complexes as catalysts. A more comprehensive global rate law was determined for this process, including a copper-dependent pathway (at very low concentrations), in addition to an independent one, both influenced by alkaline medium. The proposed mechanism, under pseudo-first order conditions, combine intramolecular electronic transfer with reduction of the copper ion by the substrate, leading to the formation of intermediary reactive species (OH•-, O2•-, H2O2, CO2•-), responsible for initiation and propagation steps, and of short chain carbonylic products (glycolate, glycerate and formiate ions). To better understanding metal-carbohydrate interactions, copper(II) complexes with simple monosacharides were isolated, and characterized by elemental and thermogravimetric analyses, and spectroscopic techniques (IR, UV/Vis, and EPR), besides Raman spectroscopy, used to investigate the binding mode of the carbohydrate moiety to the copper ion, in each one of these complexes. Additionally, two novel chiral imine copper(II) complexes, derived from aminocarbohydrate ligands, were prepared and characterized by elemental analysis, conductivity measurements, and spectroscopic techniques (IR, UV/Vis, EPR, ESI-MS and CD), and one of them was also used in biological studies. Biological activity studies were carried out with the imine and diimine copper(II) complexes derived from isatin, verifying antiproliferative effect toward some tumor cell lines (promonocite U937 and neuroblastoma SH-SY5Y). Cells treated with the most active complexes were committed by the apoptotic program, as verified by citofluorimetric assays, with the complexes interfering the cell cycle in different ways (G1, G2/M or S phase. Formation of free radicals was detected, and citoplasmatic carbonylated and glycosilated proteins inside the treated cells were determined by imunologic assays. In conclusion, these studies indicated modulation of the biological activity by the imine ligand in the copper(II) complexes, with very different antiproliferative behavior, going from undetectable activity to high efficacy. Among the most active compounds, a different specificity and action mode in both cell type could be observed, evidencing their potential application as antitumoral agents.

Metalofármacos baseados em rutênio têm se mostrado promissores com relação à atividade anticancerígena frente a diversos tipos de tumores. Nosso grupo de pesquisa dedica-se ao estudo de compostos contendo o centro dimetálico de valência mista Ru2(II,III) coordenado a ligantes derivados de Faines (Fármacos anti-inflamatórios não esteroides), tendo demostrando o potencial desses complexos frente a glioma. O entendimento do modo de ação destes complexos requer o estudo de suas interações com biomoléculas presentes no meio biológico. Neste cenário, o presente trabalho teve por objetivo investigar a interação de três complexos de dirutênio-Faines, ou RuFaines, [Ru2(ibp)4Cl], [Ru2(ceto)4Cl] e [Ru2(npx)4(H2O)2]PF6 (ibp = ibuprofenato, ceto = cetoprofenato e npx = naproxenato), e também do precursor [Ru2(O2CCH3)4Cl], RuAc, com as principais proteínas presentes no soro humano, transferrina e albumina. Os complexos foram sintetizados e caracterizados conforme metodologias desenvolvidas no grupo. A interação destes complexos com a transferrina, em suas formas apo e holo, e com a albumina foi avaliada por técnicas como espectroscopia eletrônica, dicroísmo circular, fluorescência, e realizaram-se estudos de ultrafiltração com análise do aduto formado por ICP-OES e espectrometria de massas. Além disso, fez-se um estudo de captação celular dos complexos RuFaines por células de glioma humano da linhagem U-87. Os resultados demonstraram que os complexos de dirutênio-Faines interagem com ambas as proteínas séricas (transferrina (apo e holo) e albumina), de modo semelhante, mas que é distinto daquele observado para o complexo RuAc. A presença de íons Fe(III) nos sítios específicos da transferrina não afetou a interação dos complexos RuFaines, enquanto que um comportamento diferente foi observado para o RuAc. Verificou-se que todas as proteínas avaliadas (albumina, apo-transferrina e holo-transferrina) apresentam capacidades similares de retenção dos complexos (~ 70% da quantidade de Ru adicionada inicialmente), independentemente da natureza do ligante carboxilato coordenado. Estudos de captação celular mostraram que a interação dos complexos RuFaines com a transferrina não contribuiu para modificar a capacidade de entrada desses complexos na célula, em comparação com os metalofármacos livres. Em alguns casos, inclusive, a formação de aduto com a apo-transferrina teve um efeito contrário, diminuindo a captação de rutênio. Dessa forma, concluiu-se que o ciclo da transferrina provavelmente não é a principal rota de entrada nas células para os complexos estudados. / Ruthenium metallodrugs have shown promising antitumor activity against to several tumor types. Our research group is dedicated to study compounds containing the mixed-valence Ru2(II,III) dimetallic center coordinated to NSAIDs (nonsteroidal anti-inflammatory drugs) derived ligands, and have demonstrated the potential of these complexes against glioma. The understanding of the mode of action of these complexes requires the study of their interactions with biomolecules present in biological environment. In this scenario, the present work aimed to investigate the interaction of three complexes of diruthenium-NSAIDs, or RuNSAIDs, [Ru2(ibp)4Cl], [Ru2(ceto)4Cl] and [Ru2(npx)4(H2O)2]PF6 (ibp = ibuprofenate, ceto = ketoprofenate, npx = naproxenate), and also of the precursor [Ru2(O2CCH3)4Cl], RuAc, with the major proteins present in the human serum, transferrin and albumin. The complexes were synthesized and characterized according to methods developed in our group. The interaction of these complexes with transferrin, in the two forms apo and holo, and with albumin was evaluated by techniques as electronic spectroscopy, circular dichroism, fluorescence, and ultrafiltration studies accompanied by analysis of adducts by ICP-OES and mass spectrometry. Moreover, cellular uptake studies of the RuNSAIDs complexes by U-87 human glioma cells line were performed. The results demonstrated that the diruthenium-NSAIDs complexes interact with both proteins (transferrin (apo and holo) and albumin), in a similar way, but that is distinct of that observed for the RuAc complex. The presence of Fe(III) ions in transferrin specific binding sites did not affect the interaction of the RuNSAID complexes with the protein, while a different behavior was shown by RuAc. All the proteins studied here (albumin, apo-transferrin and holo-transferrin) showed similar capabilities for retention of the complexes (~ 70 % of the initial amount of Ru added), independently of the nature of the coordinated carboxylate ligand. Cellular uptake studies showed that the interaction of the RuNSAIDs complexes with transferrin did not contribute to modify the internalization capacity of these complexes, in comparison with the free metallodrugs. In some cases, the adduct formation with apo-transferrin showed an opposite effect, leading to the decrease of ruthenium uptake. The findings led to the conclusion that transferrin cycle probably is not the main entry way to the cells for the studied complexes.

A bioatividade de complexos metálicos é de grande interesse atual na área de pesquisa de novos medicamentos. Fármacos anti-inflamatórios não-esteróides (FAINEs) são amplamente consumidos para o tratamento de doenças inflamatórias e de dor, mas seu contínuo uso pode ocasionar sérios efeitos colaterais ao trato gastrointestinal (TGI). Metalofármacos alternativos de Cu-FAINEs causam menores danos ao TGI, sem perder a atividade anti-inflamatória. No entanto, alguns destes são pouco solúveis. Adicionalmente, o desenvolvimento de materiais híbridos de FAINEs/polímeros para liberação modificada dos fármacos é interessante do ponto de vista de se aumentar a eficácia terapêutica e minimizar problemas de toxicidade. O objetivo deste trabalho foi investigar metodologia, com emprego de secagem via spray-dryer, para a preparação de materiais híbridos contendo complexos de cobre(II) com os fármacos naproxeno e sulindaco incorporados em acetato de celulose. Dois tipos de materiais híbridos foram obtidos para cada metalofármaco: CuFAINE/AC e CuFAINE/AC/OM, sendo o segundo preparado em presença de óleo mineral (OM). A caracterização destes produtos foi efetuada por análises elementares, espectroscopia vibracional no infravermelho, difratometria de raios X, análise termogravimétrica acoplada a espectrômetro de massas, ressonância paramagnética eletrônica, microscopia eletrônica de varredura e espalhamento de luz dinâmico. Os métodos utilizados levaram à interação dos metalofármacos de CuFAINEs com a matriz polimérica biocompatível de acetato de celulose, gerando materiais híbridos bioinorgânicos-orgânicos. Duas espécies de CuFAINEs: uma dimérica mantendo a estrutura original ([Cu2(FAINE)4(Laxial)2]) e outra monomérica - provavelmente do tipo ([Cu(FAINE)2(Laxial)2]) - foram identificadas nestes materiais. As partículas apresentaram tamanho nanométrico. Os resultados aqui obtidos abrem novas perspectivas para futuros estudos que visem à investigação da viabilidade de usar sistemas como estes para liberação mofidicada dos metalofármacos de CuFAINEs / The bioactivity of metal complexes has current and high interest in the field of development of new drugs. Non-steroidal anti-inflammatory drugs (NSAIDs) are widely consumed to treat inflammatory diseases and pain. The continuous use of these drugs causes serious side-effects on the gastrointestinal tract (GT). Alternative copper(II)-metallodrugs (CuNSAIDs) show lower GT side-effects without loss of anti-inflammatory activity. However, some of them are poorly water-soluble. In addition, the development of NSAID/biocompatible polymeric hybrid materials for modified-release of pharmaceuticals is of interest to increase therapeutic efficacy and to lower toxicity of drugs. The aim of this work was to investigate a methodology adding the use of spray-drying technique to prepare hybrid materials containing copper(II) complexes with naproxen and sulindac drugs incorporated into cellulose acetate (CA). Two different materials were obtained for each metallodrug: CuNSAID/CA and CuNSAID/CA/MO, being the second one prepared in the presence of mineral oil (MO). The characterization of these products was conducted by elemental analysis, vibrational spectroscopy, powder x-ray diffraction, thermogravimetric analysis coupled to mass spectrometry, electron paramagnetic resonance, scanning electron microscopy and dynamic light scattering. The developed methodologies led to the interaction between the CuNSAIDs and the polymeric matrix generating bioinorganic-organic hybrid materials. Two CuNSAIDs species: a dimeric one that kept the complex original structure [Cu2(FAINE)4(Laxial)2] and a monomeric species, ([Cu(FAINE)2(Laxial)2] were identified in these materials. The particles have shown nanometric size. The obtained results open up new perspectives for future studies with the aim of investigating the availability of using this type of system for modified release of CuNSAIDs metallodrugs.