Thesis (M.S.)--University of North Carolina at Wilmington, 2005. / Includes bibliographical references (leaves: 57-60)

This thesis includes the synthesis, characterisation, antioxidant and antimicrobial activities of Cu(II)-, Co(II)- and Co(III) complexes with N,N'-bis(3- methoxysalicylaldimine)propan-2-ol, 2-OH-oVANPN. The Schiff base ligand, 2-OHoVANPN, is derived from o-vanillin and 1,3-diaminopropan-2-ol. The o-vanillin condensed with 1,3-diaminopropan-2-ol in a 2:1 molar ratio yields this potential tetraor pentadentate ligand. The complexes synthesized are tetra (or penta or hexa) coordinated. Formation of the complexes is symbolized as follows:- MX₂ + 2-OH-oVANPN (2:1) -> [M(2-OH-oVANPN)Xn] + HnX MX₂ + 2-OH-oVANPN (2:1) -> [Mn(2-OH-oVANPN)OH] + H₂X₂ MX₂ + (o-vanillin : diaminopropanol) (1:1) -> [M(1:1)X₂] MX₂ + (o-vanillin : diaminopropanol) (1:1) -> [M₃(1:1)X₄] M = Cu(II), Co(II) or Co(III); X = Cl; n = 1, 2. Their structural features have been deduced from their elemental analytical data, IR spectral data, and electronic spectral data. With the exception of {Cu₃(C₁₁H₁₄N₂O₃)(Cl)₄(H₂O)₆}(A4), the Cu(II) complexes were monomeric with 2-OH-oVANPN acting as a tetradentate ligand. A binuclear Co(II) complex, [Co₂(C₁₉H₁₉N₂O₅)(OH)] (B1), was synthesised and the rest of the Co(II) and Co(III) complexes were monomeric with chloride ions coordinating to the metal centre in some cases. Electronic data suggest that the cobalt(II) complexes have octahedral geometries and the copper(II) complexes have square planar structures – Co(III) is likely to be octahedral. Thermal analyses, which included the copper-block-method for determining sublimation temperatures, revealed that some copper(II) and cobalt(II) complexes are hygroscopic and sublime at 200 °C and below. DSC analyses of the Cu(II) complexes gave exotherms around 300 °C for complexes K[Cu(C₁₉H₂₀N₂O₅)(OH)]·2H₂O (A1) and [Cu(C₁₁H15N₂O₃)(Cl)₂]·2H₂O (A2) and above 400 °C for [Cu(C₁₁H₁₆N₂O₃)(Cl)₂] (A3) and {Cu₃(C₁₁H₁₄N₂O₃)(Cl)₄(H₂O)₆} (A4). Antioxidant studies were carried out against the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH·). The cobalt(II) complex, [Co₂(C₁₉H₁₉N₂O₅)(OH)] (B1), which was synthesized in the presence of KOH, had no antioxidant activity, whilst the other cobalt(II) complexes, [Co(C₁₇H₁₇N₂O₅(Cl))]·1½H₂O (B2), [Co(C₁₉H₂₂N₂O₅) (Cl)₂]·5½H₂O (B3) and [Co(C₁₉H₂₂N₂O₅)(Cl)₂]·5½H₂O (B4), which were synthesised in the absence of KOH, demonstrated antioxidant activity. The latter complexes are candidates for cancer cell line testing, while [Cu(C₁₁H₁₆N₂O₃)(Cl)₂] (A3), {Cu₃(C₁₁H₁₄N₂O₃)(Cl)₄(H₂O)₆} (A4), [Co(C₁₉H₂₁N₂O₅)(Cl)₂ ]·5H₂O (C2) and [Co(C₁₉H₂₀N₂O₅)(Cl)]·3H₂O (C3) may show anticancer activity through possible hydrolysis products. Most of the complexes synthesized displayed antimicrobial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Aspergillus niger and Candida albicans. The results indicated that complexes [Cu(C₁₁H₁₆N₂O₃)(Cl)₂](A3), [Co(C₁₉H₂₂N₂O₅)(Cl)₂]·5½H₂O (B3) and [Co(C₁₉H₂₁N₂O₅)(Cl)₂ ]·5H₂O (C2) are active against the Gram-negative Ps. aeruginosa and that the ligand, 2-OH-oVANPN, did not have any activity. The same trend was observed with 2-OH-oVANPN, {Cu₃(C₁₁H₁₄N₂O₃)(Cl)4(H₂O)₆} (A4) and [Co(C₁₉H₂₀N₂O₅)(Cl)]·3H₂O (C3) against the Gram-positive S. aureus. As for activity against E. coli and C. albicans, some complexes showed more activity than the ligand. There is an observed trend here that the metal complexes are more active (toxic) than the corresponding ligand, which is in agreement with Tweedy’s chelation theory.

Orientador: Ronaldo Aloise Pilli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-18T10:51:34Z (GMT). No. of bitstreams: 1 Castro_IltonBarrosDaltrode_D.pdf: 7677893 bytes, checksum: 67cdce338d4518746ab605e0f0b6e600 (MD5) Previous issue date: 2010 / Resumo: Fostriecina (10) é um fosfato orgânico natural citotóxico isolado de Streptomyces pulveraceu e apresenta potente atividade anticâncer contra células leucêmicas (IC50=0,46 mM) e outras linhagens de células tumorais. As propriedades citotóxicas desse composto são atribuídas à inibição seletiva da proteína fosfatase 2A (IC50=3,2 mM). A fostriecina havia avançado para estudos clínicos, no entanto os estudos foram suspensos devido à instabilidade do composto, que foi atribuída à porção triênica da sua estrutura. Com base nisso, foi decidido estudar análogos de fostriecina (105a e 105b) que apresentem estabilidade química atrelada a potente atividade biológica. A estratégia de síntese proposta para a obtenção dos análogos 105a e 105b foi orientada por algumas idéias centrais que visavam preservar a integridade do anel di-hidropirano, controlar adequadamente a geometria das ligações duplas presentes na estrutura e a estereoquímica absoluta dos centros estereogênicos, sem comprometer a eficiência da rota sintética. As etapas chave na preparação dos análogos de fostriecina foram: a reação de alilação catalítica e assimétrica de Yamamoto para instalação do centro estereogênico em C5; a reação de di-hidroxilação assimétrica de Sharpless para construção do centro estereogênico em C8 e a reação aldólica para construção da ligação C10-C11. Os produtos finais da síntese, bem como os intermediários avançados da síntese, foram submetidos a três ensaios: de atividade antiproliferativa in vitro contra 10 linhagens de células tumorais, de inibição da enzima LMWPTP isolada e de viabilidade celular em linhagens que superexpressam a LMWPTP. Três compostos se mostraram bastante promissores nos ensaios, apresentando inibição superior aos controles positivos e com baixa toxicidade para células sadias. Os resultados obtidos sugerem que os compostos induzem morte celular via perturbação das vias de sinalização da enzima LMWPTP / Abstract: Fostriecin (10) is a natural cytotoxic organic phosphate isolated from Streptomyces pulveraceus which exhibits potent antitumor activity against leukemic cells (IC50=0,46 mM), among others. The cytotoxic properties of this compound are presumably linked to the selective inhibition of the phosphatase protein 2A (IC50=3,2 mM). Although fostriecin has advanced to clinical studies, these were suspended due to its chemical instability, which was assigned to the trienic moiety of the structure. Therefore it was decided to prepare fostriecin analogues (105a and 105b), which should present chemical stability and potent biological activity.The synthetic strategy proposed towards the achievement of analogues 105a and 105b was guided by a main idea that aimed to preserve the integrity of the dihydropyran ring and to properly control the geometry of the double bonds, as well as the absolute stereochemistry of the stereogenic centers, without compromising the efficiency of the synthetic route. The key steps in the preparation of the fostriecin analogues were: a Yamamoto catalytic and asymmetric allylation reaction to install the stereogenic center in C5; a Sharpless asymmetric dihydroxilation reaction to access the stereogenic center in C8; and an aldol reaction to form the bond between C10-C11. The target products, as well as advanced synthetic intermediates were submitted to three biological assays: antiproliferative activity in vitro against 10 cancer cell lines; LMWPTP isolated enzyme inhibition; and cell viability within cell lines that overexpress the LMWPTP. Three of those compounds showed promising results, exhibiting higher inhibition than the positive controls and with low cytotoxicity to healthy cells. These results suggest that these compounds induct apoptosis via disruption of the LMWPTP enzyme signaling pathways / Doutorado / Quimica Organica / Doutor em Ciências

Apesar de existir novas técnicas e múltiplas alternativas terapêuticas para o câncer de próstata localmente avançado, esta enfermidade se constitui em um grande problema de saúde pública mundial, resultando em índices significativos de morbidade e mortalidade, gerando desta forma um desafio para urologistas e oncologistas. Existem múltiplas e bem sucedidas estratégias de tratamento da doença localizada, tais como: a prostatectomia radical, a radioterapia externa conformacional, a braquiterapia e a crioablação. Em contraste, o tratamento da doença metastática e localmente avançada, freqüentemente necessita da alguma forma de bloqueio hormonal. Não existe consenso em vários aspectos da terapia hormonal para tumores localmente avançados tais como: o tipo de bloqueio androgênico a ser usado, terapia hormonal precoce ou tardia, associação com outras modalidades terapêuticas e o uso de bloqueio intermitente. Foi realizada uma revisão crítica deste tipo de tratamento, bem como as indicações atuais de bloqueio hormonal nos tumores de próstata localmente avançado. Não existem estudos prospectivos e randomizados que comparem as diversas formas de tratamento cirúrgico versus radioterápico do câncer de próstata localmente avançado. A hormonioterapia adjuvante à prostatectomia radical, na doença localmente avançada, parece reduzir a progressão tumoral bioquímica, porém, não há estudo que evidencie melhora na sobrevida livre de metástase ou na sobrevida global. O bloqueio androgênico neoadjuvante à prostatectomia radical aumenta a proporção dos pacientes com doença órgão-confinada e margens cirúrgicas negativas, porém sem efeito nas taxas de falha bioquímica do tratamento. A terapia hormonal adjuvante à radioterapia em pacientes portadores de câncer de próstata localmente avançado oferece vantagens na sobrevida global. A terapia hormonal neoadjuvante à radioterapia, em estudos multicêntricos e randomizados, resulta em melhor controle local do tumor bem como prolonga a sobrevida doença-específica. Não há, porém evidência de melhora na sobrevida global. O tratamento por tempo prolongado com bloqueadores hormonais adjuvante à radioterapia mostrou-se superior em relação à sobrevida global e sobrevida livre de doença quando comparado a um período curto de bloqueio, principalmente em pacientes com tumores indiferenciados (Gleason 8-10). Os análogos LHRH, orquiectomia ou o dietilestilbestrol se mostraram como opções de monoterapia, igualmente eficazes, para os pacientes que iniciam terapia hormonal de primeira linha, no tratamento da doença localmente avançada. Não existe evidência que justifique o bloqueio androgênico máximo como terapia hormonal de primeira linha ao invés de monoterapia. Existem vantagens potenciais na qualidade de vida e nos custos do tratamento quando realizada a ablação intermitente, mas a sua eficácia a longo prazo necessita ser confirmada / Despite new techniques and multiple therapeutic alternatives, locally advanced prostate cancer is a serious public health problem, resulting in significant morbidity and mortality rates, that remains a great challenge for urologists and oncologists. Several therapeutic strategies to treat localized prostate cancer have been successful such as conformational external beam radiation therapy, brachytherapy and cryoablation. In contrast, treatment of metastatic and locally advanced tumors may often involve androgenic suppression. However, there are no consensus on several aspects of hormonal therapy for locally advanced tumors such as the type of antiandrogenic drug to be used, early versus delayed hormonal therapy, association with other therapeutic modalities and the use of intermittent blockade. We set out to critically review important aspects and current indications of hormonal blockade in the locally advanced prostate tumors. There are no prospective and randomized study that compares current forms of surgical treatment versus radiation therapy of locally advanced prostate cancer. After radical prostatectomy, adjuvant hormonal therapy in the locally advanced disease reduces biochemical failure rates, although no benefit has been shown regarding metastatic free survival or overall suvival. Neoadjuvant androgen blockade enhances the proportion of patients with organ-confined disease and negative surgical margins but no benefit is seen regarding biochemical free recurrence. Neoadjuvant hormonal therapy to the radiotherapy improves local tumor control as well as it prolongs the diseasespecific survival, although there are no survival advantage. Adjuvant hormonal therapy offers overall survival advantage in patients with locally advanced prostate cancer treated with radiotherapy Long term adjuvant hormonal blockade offers survival benefit for patients with high Gleason score (8-10). LHRH analogues, bilateral orquiectomy and dietilestilbestrol were shown are equally effective as adjuvant therapy for patients with locally disease advanced. There are evidences that maximum androgenic blockade are not more efficient than monotherapy. Potential quality of life and costs advantages of intermittent ablation could be considered an alternative treatment for this group of patient

No presente estudo realizou-se a análise custo-efetividade das terapias antineoplásicas IFL (irinotecano, 5-fluorouracil e leucovorin) e IFL+BV (IFL associado ao bevacizumabe) empregado no tratamento do câncer colorretal metastático em primeira linha. Estimou o custo direto de medicamentos, materiais e recursos humanos. A efetividade dos protocolos foi medida pela proporção de pacientes livre de progressão de doença. Os dados relativos aos custos de materiais e medicamentos foram obtidos a partir de tabelas de preços que regulamentam o mercado hospitalar. Os dados concernentes à efetividade foram obtidos através da literatura científica. Utilizou-se o modelo de análise de decisão para estimar o custo total da terapia antineoplásica. Os resultados mostraram que o protocolo IFL apresentou a melhor relação custo-efetividade durante todo o tempo de seguimento, ou seja, o menor custo por unidade de efetividade, que no 10º mês foi de R$ 180.619,46. A análise de sensibilidade mostrou que esta conclusão foi robusta. Essas análises farmacoeconômicas apontaram que a seleção do protocolo antineoplásico depende do custo e efetividade, mas, sobretudo da relação custo-efetividade que permite saber o custo estimado por unidade de sucesso. / In this paper a cost–effectiveness analyses was done of the antineoplasics therapies IFL (irinotecan, 5- fluorouracil and leucovorin) and IFL + BV (IFL associated to bevacizumab) used as metastatic colorectal cancer as first line treatment. It has estimated the cost of medications, materials and human resources. The effectiveness of the protocols was measured through the proportion of patients that were free from the illness progression. Data regarding material cost and medication were obtained by price tables that regulate Hospital market. Data relating to effectiveness were obtained through scientific literature. We utilized the decision analysis model to estimate the total cost of the antineoplasics therapy. The results showed that the IFL protocol presented a better cost–effectiveness relationship during the whole period following, that is, the lowest cost per effectiveness units, that on the 10th Month was R$ 180.619,46. The sensitivity analysis showed that this conclusion was strong. These pharmacoeconomic analyses pointed to the fact that antineoplasics protocol selection depends on cost and effectiveness, but, above all on the cost–effectiveness relation which allows us to know the estimated cost per successful unit.

OBJETIVO: Avaliar a eficácia e segurança da imunoterapia tópica com imiquimode creme 5% no tratamento do carcinoma basocelular nodular periocular. MÉTODOS: Pacientes com carcinoma basocelular confirmado por biopsia e com contraindicação clínica para a cirurgia reconstrutiva devido ao alto risco ou que recusaram a cirurgia por razões estéticas ou fobia foram incluídos no estudo. O tratamento foi iniciado após treinamento do paciente e de acompanhante. A posologia foi de 5 vezes por semana por 8 a 16 semanas. Acompanhamento quinzenal foi realizado durante a vigência do tratamento com questionário, exame biomicroscópico, medida da acuidade visual e documentação fotográfica. As características clínicas das lesões foram mensuradas através do software ImageJ. Após 12 semanas do fim da terapia, uma nova biópsia na região da lesão foi guiada por fotografia. O seguimento dos pacientes foi semestral, após fim do tratamento, com biópsias anuais da região até o presente momento. RESULTADOS: 19 foram tratadas. A taxa de cura histológica foi de 89,5% após três meses do final do tratamento, e de 84,2% nos três anos de seguimento (39,5 meses). A taxa de cura histológica em três anos foi de 100% para lesões menores que 10 mm, e de 81,8% para lesões maiores que 10 mm. De uma forma geral, os efeitos colaterais da medicação foram mais frequentes durante as oito primeiras semanas de tratamento. Quanto menor foi a distância da lesão à margem palpebral, maior foi chance de o paciente desenvolver ectrópio no tratamento (p = 0,045). Assim, como quanto maior foi a inflamação, maior foi a chance de desenvolver ectrópio, dor e edema durante o tratamento (p = 0,017, p = 0,016 e p = 0,044, respectivamente). CONCLUSÕES: Imiquimode creme 5% mostrou-se eficaz para o tratamento alternativo do carcinoma basocelular periocular, principalmente em lesões menores que 10 mm. Em adição, demonstrou um interessante efeito neoadjuvante sobre as lesões maiores que 10 mm que não foram curadas. Mostrou-se um tratamento seguro; entretanto, um cuidado maior deve ser dado às lesões próximas à margem palpebral devido ao maior risco de complicações e desenvolvimento de ectrópio / Objective: to evaluate the efficacy and safety of topical administration of 5% imiquimod cream in the treatment of periocular nodular basal cell carcinoma (BCC). Methods: Patients with periocular nodular basal cell carcinoma confirmed by biopsy and clinical contraindication to reconstructive surgery due to high risk or who decline surgery for aesthetic reasons or phobia were included in the study. The medication was applied once a day, five days a week for 8-16 weeks. Treatment was initiated after provision of patient and caretaker training. During treatment, patients were followed up twice a month with questionnaires, biomicroscopic examinations, measurement of visual acuity and photographic documentation. The clinical characteristics of the tumors were registered with the software ImageJ. Twelve weeks after the end of treatment, an image-guided biopsy of the tumor site was performed. Patients have since been attending follow-up visits every six months, and biopsies of the region are performed annually. Results: 19 tumors were treated with imiquimod. The average histological cure rate was 89.5% after 3 months at the end of the treatment and 84.2% after 3 years of follow-up (39.5 months). The 3-year histological cure rate was 100% for smaller tumors and 81.8% for larger tumors (>10 mm). In general, drug-related side effects were more frequent during the first 8 weeks of treatment. The smaller the distance between tumor and lid margin, the greater the probability of developing ectropion during treatment (p=0.045). Likewise, the more severe the inflammation, the greater the probability of developing ectropion, pain and edema during treatment (p=0.017, p=0.016 and p=0.044 respectively). Conclusion: Topical administration of 5% imiquimod cream was found to be an efficacious and relatively safe alternative treatment for periocular BCC, especially for tumors smaller than 10 mm, with interesting neoadjuvant effects on uncured tumors larger than 10 mm. However, special care is required when treating tumors near the eyelid margin due to the risk of complications and development of ectropion

La plupart des molécules utilisées en chimiothérapie conventionnelle, bien qu’ayant des cibles moléculaires différentes, induisent dans la majorité des cas une mort cellulaire par apoptose. Or, de plus en plus de chimiorésistances se rencontrent au niveau des cellules cancéreuses vis-à-vis de ce type de molécules. Face à cette situation il devient urgent de trouver des molécules ayant des mécanismes d’action différents et capables de court-circuiter spécifiquement les mécanismes de résistance des cellules cancéreuses. <p>La stratégie mise en place lors de ce travail a été de partir d’une molécule naturelle issue d’un extrait de la racine de Glycyrrhiza glabra qui présentait déjà une activité anti tumorale marquée. L’intérêt du travail a été de dériver l’acide 18β-glycyrrhétinique de manière originale afin de potentialiser son effet anticancéreux, notamment vis-à-vis de huit lignées cellulaires présentant des résistances plus ou moins marquées aux stimuli pro-apoptotiques. Ainsi après avoir caractérisé la pureté et la stabilité de cette série de nouvelles molécules, nous avons retenu les dérivés les plus intéressants en termes d’inhibition in vitro de la prolifération cellulaire. Sur base de ce premier choix, nous avons investigué des cibles spécifiques décrites dans la littérature pour les hémidérivés de l’acide 18β-glycyrrhétinique :le protéasome 26S et le récepteur nucléaire PPARγ. Cette étude nous a permis de retenir un dérivé en particulier capable d’inhiber à 50% les trois sites catalytiques du protéasome sans toutefois inhiber PPARγ :le N-(2-{3-[3,5-bis(trifluoromethyl)phenyl]ureido}ethyl)-glycyrrhetinamide (6b). Nous avons ensuite évalué ce composé sur un ensemble de 333 kinases afin de déterminer un profil antitumoral plus large pour ce type de molécule. <p>Le profil pharmacologique in vitro de ce dérivé de l’acide 18β-glycyrrhétinique nous a amenés à étudier son comportement in vivo chez la souris saine. A cette fin, une étude de préformulation nous a permis de définir une formulation galénique optimale pour ce composé, la nanoémulsion qui a servi à déterminer une dose maximale tolérée (indice DMT) par la souris saine. Nous avons ensuite travaillé à une dose non toxique pour déterminer le profil pharmacocinétique plasmatique chez la souris saine, par voie d’administration intraveineuse et par voie orale. <p>Les conclusions de cette étude nous montrent que le dérivé de l’acide 18β-glycyrrhétinique que nous avons mis au point présente de remarquables caractéristiques pharmacologiques in vitro et un comportement in vivo proche de la molécule naturelle. Des études d’activité in vivo devraient débuter prochainement.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

The search for more effective treatment strategies in melanoma led to many new innovative approaches aiming at different molecular targets. Chemotherapy still remains the most effective treatment and many efforts are put in order to improve targeting and delivery of the chemotherapeutic agents. Among these, peptide conjugates of anticancer drugs were designed to increase stability, cell penetration, specificity and accumulation in cancer cells. We as well as others evaluated such a conjugate, termed PSF (L-prolyl-m-L-sarcolysyl-L-p-fluorophenylalanine-ethylester) in terms of its cytotoxicity in vitro and in vivo using a human melanoma tumor as a model, its stability, transport, and metabolisation. <p>By comparing the cytotoxicity of PSF and melphalan towards different cancer primary melanoma cell cultures, we noticed some interesting observations: PSF displayed the same toxicity pattern both in short (2h) and long term (24h) cell exposures whereas melphalan and m-sarcolysin needed long term exposure to reach the same toxicity. This could indicate that PSF very quickly penetrates the cells in accordance with what has been shown with red blood cells (RBCs). PSF has shown a much better and quicker penetration into the cells in vitro as compared to melphalan. <p>In this present work, the cytotoxic effect of PSF was further evaluated in vivo using a standardized nude mice tumor model bearing a human melanoma. First, the acute toxicity in rats and mice and the maximum tolerated dose were determined. After a dose-escalation study one dose was singled out and tested as a single dose and as a fractionated dose. PSF was able to reach the tumor site and a dose-response relationship was observed. The IP administration of fractionated doses of PSF had significantly better effect on tumor growth inhibition, regression and regrowth than single dose administration and this without any evidence for general toxicity monitored by animal weight loss. We also compared the efficacy of PSF to its parent drug m-sarcolysin, melphalan and cyclophosphamide and observed that PSF was much more active than both melphalan and m-sarcolysin at the same molar doses.<p>Body distribution of the 14C-labelled PSF revealed ratios of 2.4 and 1.5 compared to muscle tissue for the two melanoma tumors evaluated with no significant and stable accumulation in any vital organ. The amount of tracer was still high in the blood after 24 hours explaining the high radioactivity in the kidney and partly in the liver. Interestingly, the spleen had an unusual high radioactivity uptake reflecting the exceptional binding of the tracer to blood cells (BC), while the pancreas very high load was an indicator of protease-mediated specific delivery and strongly support our hypothesis elaborated on the basis of in vitro results. <p><p>Our in vitro data point to a particular mechanism of action of PSF based on the transport of PSF through the body by the rapid binding to blood cells and the delivery at the tumor site by the subsequent release of its active metabolites due to cleavage by tumor-associated proteases.<p>Concerning the binding of PSF to membranes and its transport the following observations were made: while PSF was stable in human plasma, it disappeared very quickly in whole blood along with the generation of a main metabolite: m-sarcolysin. The presence of BC membranes was required for both binding and generating the metabolites. Binding to natural or artificial membranes was achieved and only competition with melanoma cells or proteolytic enzymes such as dispase, led to the generation of active metabolites. The different metabolites were isolated using preparative LC and were then identified using Electrospray Ionisation Mass Spectrometry (ESI). Three metabolites, of which m-sarcolysin was the main one, were identified all bearing the chloroethyl alkylating group. <p>Enzymatic catalysis was further supported by a set of experiments where the enzymatic activity was non-specifically and specifically inhibited. In order to look at the effect of extracellular matrix proteases on PSF, three representatives of ECM proteases were incubated with PSF: collagenase A had no effect, but both dispase and trypsine were able to process PSF. <p>The following data indicate the higher processing of PSF in the presence of cells with a higher proteolytic activity and thus the delivery of the blood cell-bound PSF. When comparing BC with melanoma cells (MC), the latter showed a higher ability to bind and process PSF both by membrane-associated and most interestingly soluble proteases. A lot of families of enzymes are reported to be overexpressed by melanoma cells including: metalloproteases, cysteine cathepsins, serine proteases and aminopeptidases. All the melanoma cells and cell lines evaluated were able to generate PSF active metabolites. <p>To identify the families of enzymes expressed on the membrane of melanoma cells that might be involved in the mechanism of action of PSF, we performed 2D-gel electrophoresis on their membrane extracts. The 2D-gels experiments revealed the presence of proteins compatible with enzymes known to be important in melanoma and further work is needed to identify the individual enzymes involved by using mass spectrometry and Western blotting. <p><p>Both our in vitro and in vivo findings strongly suggest that not only melanoma tumor cells and tumor sites but other types of tumors as well may be targets for the toxic activity of PSF owing to their much higher load in proteolytic enzymes that are closely related to their invasive potential. The transport of PSF by the blood cells and the release of its metabolites at the tumor site result in a low amount of drug in its free soluble form within the blood and this may explain the relatively lower side-effects observed. PSF is thus expected to have a much better therapeutic index than conventional alkylating agents. This original mechanism of drug delivery may well be extended to other cancer and non-cancer drugs than alkylating agents.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Avec environ 700 000 décès en 2008, le carcinome hépatocellulaire se situe au 3ème rang de la mortalité par cancers dans le monde. La chimioembolisation est le traitement recommandé chez les patients atteints d’un carcinome hépatocellulaire de stade intermédiaire B de la classification Barcelona Clinic Liver Cancer. Cette technique de radiologie interventionnelle consiste en l’injection intraartérielle d’un agent anticancéreux à l’aide d’un vecteur (lipiodol ou microsphères d’embolisation) complétée par une occlusion artérielle lorsque le lipiodol est utilisé. La médiane de survie des patients traités par chimioembolisation pour un carcinome hépatocellulaire n’excède pas 2 ans et il n’existe aucun consensus sur la procédure optimale.L’objectif de notre travail est d’essayer d’améliorer la procédure de chimioembolisation en optimisant d’une part l’agent anticancéreux et d’autre part, son vecteur.Il a été démontré au cours d’un travail de sélection in vitro, que l’idarubicine est l’agent anticancéreux le plus cytotoxique sur 3 lignées humaines de carcinome hépatocellulaire. Cette anthracycline présente une cytotoxicité supérieure à 10 autres agents anticancéreux dont ceux utilisés en pratique clinique pour la chimioembolisation des carcinomes hépatocellulaires.L’essai de chimioembolisation de phase II randomisé LIPIOAMIO a montré que l’addition d’amiodarone utilisé pour stabiliser une émulsion à base de lipiodol et d’anthracycline n’augmente pas signicativement la survie des patients atteints d’un carcinome hépatocellulaire non résécable non métastatique. Nous avons par ailleurs montré que l’idarubicine était chargeable et donnait une solution stable plusieurs mois avec les microsphères d’embolisation DC Bead™. Un essai de phase I est en cours pour déterminer la dose limitante de l’idarubicine administrée dans une solution de microsphères DC Bead™ au cours d’une séance de chimioembolisation chez des patients atteints d’un carcinome hépatocellulaire non résécable, non métastatique. Quelques résultats préliminaires de cet essai sont présentés dans le manuscrit. / With 700,000 deaths in 2008, hepatocellular carcinoma is the 3rd most common cause of cancer-related death worldwide. Transarterial chemoembolization is the standard treatment for intermediate-stage hepatocellular carcinoma. This intraarterial treatment is performed by injecting an anticancer drug carried by ethiodized oil or by drug-eluting beads and followed by the occlusion of the artery when ethiodized oil is used. Median survival of patients remains < 2 years, and there is no consensus about the optimal treatment regimen. The aim of our work was to improve the efficacy of transarterial chemoembolization in optimizing the anticancer drug and its carrier.We have demonstrated that idarubicin was the most cytotoxic anticancer drug in an in vitro screening study of 11 anticancer drugs on 3 human hepatocellular carcinoma cell lines. Idarubicin was more cytotoxic in our experiment than the anticancer drugs which are currently used for transarterial chemoembolization of hepatocellular carcinoma.The randomized LIPIOAMIO phase II trial has shown that the addition of amiodarone to stabilize an emulsion composed of an anthracycline and of ethiodized oil injected for transarterial chemoembolization does not improve significantly survival of patients with a non resectable, non metastatic hepatocellular carcinoma. We have also demonstrated that idarubicin could be loaded in drug-eluting DC Bead™ and that the resulting solution was stable during several months.We designed the dose-escalation IDASPHERE phase I trial to determine the limiting dose of idarubicin administred in a solution of drug-eluting DC Bead™ during a transarterial chemoembolization session in patients with non resectable, non metastatic hepatocellular carcinoma. First results of the trial are presented in the manuscript.

<p>Glavni problem u lečenju karcinoma dojke je kako na osnovu kliničke klasifikacije i morfolo&scaron;kih osobina tumora predvideti njegovo dalje pona&scaron;anje. Vrlo često ni kombinacija standardnih prognostičkih faktora ne daje odgovor o potrebi davanja adjuvantne hemioterapije. U cilju sprovođenja adekvatne dalje terapije karcinoma dojke i otkrivanja agresivnih tipova tumora, a nakon hirur&scaron;kog lečenja, postoji stalna potreba za pronalaženjem novih pokazatelja pomoću kojih bi se identifikovale bolesnice koje imaju povećan rizik od razvoja relapsa bolesti. Ciljevi ove studije su bili da se odredi učestalost ekspresije androgenih receptora (AR) u infiltrativnom duktalnom karcinomu dojke. Da se utvrdi povezanost ekspresije AR i kliničko-patolo&scaron;kih prognostičkih faktora u infiltrativnom duktalnom karcinomu dojke. Odnos ekspresije AR i ekspresije estrogen receptora (ER), progesteron receptora (PR) i humanog epidermalnog faktora rasta (HER-2). Da se proceni povezanosti pozitivne ekspresije AR, kao i odnosa AR/ER, sa odgovorom na primenjenu hormonsku terapiju kod ER pozitivnih bolesnica. Da se proceni povezanost ekspresije AR, kao i odnosa AR/ER, sa kliničkim tokom bolesti: pojavom recidiva, metastaza, kao i smrtnim ishodom u toku petogodi&scaron;njeg perioda praćenja pacijentkinja. Istraživanjem je obuhvaćeno oko 200 pacijentkinja obolelih od infiltrativnog duktalnog karcinoma dojke, koje su operisane na Institutu za onkologiju Vojvodine u periodu 2010-2012. godine. Pacijentkinje su odabrane metodom slučajnog izbora. Ne postoji statistički značajna razlika između kliničko patololo&scaron;kih faktora i ekspresije androgenih receptora. Kod pacijentkinja sa infiltrativnim duktalnih karcinomom dojke koje su ER-/AR+ nije pokazana statistički značajna razlika u HER2 proteinskoj ekspresiji. Učestalost receptora za progesteron, estrogen, HER2, Ki-67, tripl negativne ćelija ne karakteri&scaron;u prisustvo androgenskih receptora Nije dokazana statistička značajnost za prvi i drugi stadijum bolesti duktalnog invazivnog karcinoma dojke kada se uzme u obzir kraće vreme preživljavanja kod pacijentkinja koje su primale hormonoterapiju. Statistički značajno kraće vreme preživljavanja pokazano je za treći stadijum bolesti kod pacijentkinja koje su AR i ER (&ge; 2) u odnosu na pacijentkinje kod kojih je odnos AR/ER &lt; 2, čime je za treći stadijum bolesti dokazana inicijalna hipoteza . Analize u prikazanom istraživanju nisu pokazale statističku značajnost kada se porede učestalost relapsa i smrtnog ishoda kada se posmatraju pacijentkinje sa AR pozitivnim i AR negativnim infiltrativnim duktalnim karcinomom dojke. Pokazana je statistički značajna razlika u učestalosti smrtnog ishoda između pacijenatkinja koje su lečene i inhibitorima aromataze i tamoksifenom. Zaključci ove studije bi mogli biti osnova za preporuku da se utvrđivanje ekspresije AR kod karcinoma dojke uvrsti u rutinsku praksu i sadržaj patohistolo&scaron;kog nalaza. Određivanje odnosa ekspresije AR i ER u grupi ER pozitivnih bolesnica moglo bi poslužiti kao vodič za primenu konvencionalne hormonske terapije ili, s druge strane, preporuka za terapiju antiandrogenima, sa ciljem da se izborom novih terapijskih modaliteta pobolj&scaron;a efikasnost lečenja bolesnica sa karcinomom dojke.</p> / <p>The main problem in the treatment of breast cancer is how to predict its future behavior based on the clinical classification and morphological characteristics of the tumor. Very often even a combination of standard prognostic factors does not answer the need for adjuvant chemotherapy. In order to conduct adequate further breast cancer therapy and to detect aggressive tumor types, and following surgical treatment, there is a continuing need to find new indicators to identify patients at increased risk of relapse. The objectives of this study were to determine the frequency of androgen receptor (AR) expression in infiltrative ductal breast cancer. To determine the association between AR expression and clinical-pathological prognostic factors in infiltrative ductal breast cancer. Relationship between AR expression and expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor (HER-2). To evaluate the association of positive AR expression, as well as the AR / ER ratio, with response to hormone therapy in ER positive patients. To evaluate the association of AR expression, as well as the relationship of AR / ER, with the clinical course of the disease: onset of relapse, metastasis, as well as fatal outcome during the 5-year follow-up period. The study included about 200 patients suffering from infiltrative ductal breast cancer, operated on at the Institute of Oncology of Vojvodina in the period 2010-2012. years. Patients were selected by random selection. The results there is no statistically significant difference between clinically pathologic factors and androgen receptor expression. No statistically significant difference in HER2 protein expression was shown in patients with infiltrative ductal breast cancer who are ER- / AR +. The frequency of progesterone receptors, estrogen, HER2, Ki-67, tripl negative cells do not characterize the presence of androgen receptors. No statistical significance was demonstrated for the first and second stages of ductal invasive breast cancer when considering shorter survival times in patients receiving hormone therapy. A statistically significant shorter survival time was shown for the third stage of disease in patients with AR and ER (&ge; 2) compared to patients with an AR / ER ratio of &lt;2, thus proving an initial hypothesis for the third stage of disease. The analyzes in the study presented showed no statistical significance when comparing the incidence of relapse and death when looking at patients with AR positive and AR negative infiltrative ductal breast cancer. There was a statistically significant difference in the incidence of death between patients treated with both aromatase inhibitors and tamoxifen. Conclusions of this study could be the basis for recommending that the determination of AR expression in breast cancer be incorporated into the routine practice and content of pathohistological findings. Determining the ratio of AR and ER expression in a group of ER-positive patients could serve as a guide for the administration of conventional hormone therapy or, on the other hand, a recommendation for anti-androgen therapy, with the aim of improving the effectiveness of breast cancer treatment in the choice of new therapeutic modalities.</p>