In dieser Arbeit wurde die zelluläre Pharmakokinetik mit den Teilaspekten Invasion, Evasion und intrazellulärer Verteilung sowie die Apoptoseinduktion als Parameter der Pharmakodyna- mik von zwei Anthrazyklinen, dem freien und liposomal verkapseltem Daunorubicin unter- sucht. Die Versuche wurden anhand einer T-lymfatischen Zelllinie, den CEM-Zellen, durchgeführt. Mittels Durchflußzytometer und konfokaler Lasermikroskop wurde die intrazelluläre Fluoreszenz gemessen, die der intrazellulären Konzentration entsprach. Es zeigte sich, dass freies Daunorubicin anfangs deutlich schneller in die Zellen einströmte als liposomal verkapseltes Daunorubicin und früher die maximale Konzentration erreichte. Über eine längere Versuchszeit kam es aber zu einer Angleichung der maximal erreichten Konzen- trationen. Der Invasionsverlauf von Daunoxome verlief sigmoidförmig, während Daunorubi- cin einer Sättigungskinetik folgte. Der Invasionsverlauf beider Anthrazykline war sowohl zeit- als auch konzentrationsabhängig. Die Versuche zur intrazellulären Verteilung zeigten, dass sich beide Stoffe nach drei Stunden Inkubationszeit vom Zytoplasma in den Kern verteilten. Daunorubicin erreichte sehr schnell seine maximale Fluoreszenz im Kern. Bei Daunoxome ließ sich auch nach sechs Stunden Inkubation eine weitere Zunahme der Fluoreszenz messen. Die Untersuchungen zur Apoptoseinduktion unterstützten die Aussagen zur Invasion. Dauno- rubicin induzierte zu Anfang deutlich schneller Apoptose als Daunoxome. Über den gemessenen Versuchszeitraum kam es aber zu einer Angleichung aller Apoptoseraten. Auch hier zeigte sich eine Zeit- und Konzentrationsabhängigkeit. Die Ergebnisse der Evasionsversuche zeigten, dass Daunorubicin biphasisch und Daunoxome monophasisch ausströmte. Zusammenfassend kann gesagt werden, dass freies Daunorubicin initial eine bessere zelluläre Pharmakokinetik und damit eine höhere Zytotoxizität aufweist als liposomal verkapseltes Daunorubicin. Über die Zeit kommt es allerdings zu einer Angleichung der Zytotoxizität. Damit ist Daunoxome auf zellulärer Ebene mindestens genauso wirksam wie Daunorubicin. / We studied the cellular pharmacokinetics, including uptake, intracellular distribution and efflux, and the induction of apoptosis as a parameter of pharmacodynamics of the two anthracyclines, free and liposomal encapsulated daunorubicin. We used a flowcytometer and a confocal lasermicroscope to measure the intracellular fluorescence in CEM-cells, corresponding to the intracellular concentration of the drugs. Free daunorubicin invaded initially the cells much quicker than liposomal encapsulated daunorubicin and attained earlier the maximum concentration. After the examined time daunoxome achieved the same maximum concentration as daunorubicin. The invasion of liposomal encapsulated daunorubicin followed a sigmoid course, while free daunorubicin followed a saturation kinetic. It was shown that the uptake of both anthracyclines was time- and concentration-dependent. The examinations about the intracellular distribution showed, that both drugs accumulated in the nucleus after three hours of incubation. Daunorubicin attained quickly the maximum fluorescence there, while daunoxome increased slowly for the next six hours. The results of the apoptosis induction correlated to the results of the uptake experiments. Free daunorubicin induced initially quicker apoptosis than liposomal encapsulated daunorubicin. At the end of the measured time all the apoptosis rates of both drugs appeared to be equal. It was determined that the induction of apoptosis also is time- and concentration-dependent. The efflux of daunoxome was monophasic in contrast to a biphasic decline of daunorubicin. These results indicate that free daunorubicin has improved initial cellular pharmacokinetics and therefore enhanced cytotoxicity compared with liposomal encapsulated daunorubicin. But over the examined period both got equal cytotoxicity. Therefore daunoxome is on the cellular basis at least as effective as daunorubicin.

Adriamycin is a first line cancer treatment drug for breast cancer and many soft tissue carcinomas. Adriamycin is a highly effective anti-cancer drug. It has a fused ring system that is planar, hydrophobic and electron rich. These features allow intercalation into DNA which, along with its topoisomerase inhibition, results in its anti-cancer effectiveness. Despite its success, its use is limited by a dose-dependent cardiotoxicity. Adriamycin forms tight complexes with iron in varying iron:drug ratios. The chelated complex [Fe+3-Adriamycin (1:2)] oxidatively cleaves DNA via the generation of reactive oxygen species (ROS). Enzymes associated specifically with heart mitochondria increase ROS formation explaining why adriamycin is selectively cardiotoxic. Pretreatment of mice with the neural hormone melatonin eliminates the cardiotoxic effectiveness of adriamycin therapy without reducing the drugÆs antitumor effect (Wahab et al. 2000, Tumori 86: 157-162). This has recently been verified in human cancer patients (Lissoni et al. 1999, European Journal of Cancer 35: (12) 1688-1692). Melatonin is soluble in both lipid and aqueous environments and has no known side-effects. This study analyzes the mechanistic features of DNA damage by Fe+3-Adriamycin and how melatonin ameliorates this damage. An Fe+3-Adriamycin (1:2) complex + glutathione cleaves DNA; this oxidative DNA cleavage does not occur with adriamycin +/- glutathione or with FeADR in the absence of glutathione. Melatonin reduces this oxidative DNA cleavage by 67%. Using a supercoiled-to-nicked-circular-conversion assay in conjunction with spectroscopic analyses give results revealing : 1) Fe+3-Adriamycin (1:2) complex + glutathione forms a reactive intermediate. 2) Melatonin protects the DNA from cleavage by this reactive intermediate. 3) Enzymatic assays show that H2O2 is the primary ROS formed with downstream production of the hydroxy radical via the Fenton reaction. 4) Fe+3-Adriamycin (1:2) intercalates into DNA to the same degree as uncomplexed adriamycin. 5) Melatonin also binds to DNA but not by intercalation. These experiments indicate that the cardioprotective effect of melatonin in adriamycin therapy may stem from melatoninÆs interaction with a reactive intermediate from Fe+3-Adriamycin(1:2) complex + glutathione and/or a direct interaction of melatonin with DNA.

Insensitivity to chemotherapy is an ongoing issue in cancer treatment, one that appears to be highly dependent on patient-specific variations. It has been shown clinically that while a subset of patients will successfully respond to a particular chemotherapeutic regimen, there exists another subset of patients who when exposed to the same course of therapy will remain resistant to treatment or exhibit signs of relapse after treatment has been administered. This discrepancy raises interesting questions regarding the role that patient-specific variations play in controlling the efficacy of chemotherapy treatment regimens. Doxorubicin (Dox) is a common chemotherapeutic agent used in the treatment of a variety of solid tumors and leukemias and resistance to Dox treatment is a major issue in cancer chemotherapy, oftentimes leading to patient relapse. To gain a deeper understanding of the processes that influence Dox resistance, we must first understand the mechanisms that underlie and contribute to Dox's toxicity. To this end, the metabolic reactions that activate Dox have been implicated as major determinants of Dox cytoxicity and as possible factors that control Dox resistance in cancer cells. There are several lines of evidence that redox-dependent metabolism plays a large role in Dox toxicity. The Dox bioactivation network is comprised of a system of reduction/oxidation (redox) reactions that lead to the formation of toxic Dox metabolites and reactive oxygen species (ROS). Moreover, multi-drug resistant acute lymphoblastic leukemia cells derived from relapsed patients have elevated levels of the antioxidant glutathione and show insensitivity to Dox treatment. The redox dependence of Dox bioactivation, the understanding that Dox treatment generates ROS, and the evidence that Dox resistant cells exhibit increased antioxidant capacity, suggest the possibility that redox pathways modulate the efficacy of Dox treatment in cancer cells. The overall objectives of the proposed dissertation, therefore, were to investigate how the redox properties of the Dox bioactivation network influence Dox toxicity in acute lymphoblastic leukemia cells, and to provide evidence that cell-specific variations in the intracellular levels of these redox components influences the degree to which Dox treatment will induce cancer cell death. The significant findings of this study are that the redox reactions involved in Dox metabolism are dual-natured, containing a toxicity-generating module characterized by nicotinamide adenine dinucleotide phosphate (NADPH)-dependent Dox reductive conversion, as well as an ROS signal-generating module characterized by NADPH- and oxygen-dependent Dox redox cycling. The balance between the coupled redox reactions that comprise the toxicity- and ROS signal-generating modules of Dox bioactivation determines the sensitivity-phenotype of leukemia cells and phenotypic changes in the Dox-sensitivity of leukemia cells can be induced by the successful modulation of the Dox bioactivation network through the pharmacological inhibition of NADPH in a concentration- and cell type-dependent manner. This study highlights the importance of the intracellular redox network in controlling chemotherapy-induced ROS. The unequal distribution in antioxidant burden across the various intracellular antioxidant enzymes suggests a significant role for NADPH supply, as controlled by the enzyme glucose-6-phosphate dehydrogenase (G6PD), to the intracellular ROS buffering capacity of cells during instances of oxidative stress. Changes in G6PD activity were shown to promote protein-S-glutathionylation during oxidative stress conditions, thereby implicating G6PD in the modulation of redox-sensitive signal transduction pathways. The intracellular glutathione redox balance, a measure of the intracellular redox environment, can effectively regulate Dox-induced NF-κB signal transduction in leukemia cells. The systematic modulation of intracellular glutathione redox balance in leukemia cells by N-acetylcysteine (NAC) revealed an important role for protein S-glutathionylation mechanisms in the control of NF-κB signal transduction induced by Dox treatment. These findings identify the glutathione redox network as a potential therapeutic target for the systematic modulation of Dox sensitivity in cancer cells and elucidate the complex role that antioxidants such as NAC can play in modulating the effectiveness of Dox chemotherapy treatment regimens. Lastly, this study highlights the need for and the capacity of computational models to accurately describe the complex redox-reactions that contribute to Dox metabolism in leukemia cells. This study is groundbreaking in its use of computational modeling to analyze reversible electron transfer events between proteins using mass-action kinetics. The models developed in this study can accurately explain cytosolic doxorubicin bioactivation, intracellular hydrogen peroxide clearance, and kinase-specific S-glutathionylation, thereby showing that the use of comprehensive and/or relatively simple computational models can provide semi-quantitative predictions about the behavior of redox systems in mammalian cells as they relate to Dox-induced toxicity and Dox-induced cell signaling.

Reproduction de : Thèse doctorat : Pharmacie. Biochimie et biologie moléculaire : Reims : 2004. / Bibliogr.

O Câncer de mama é um dos problemas de saúde pública mais importantes em nosso país. São estimados, para 2010, 49.400 novos casos de câncer de mama no Brasil, com um risco estimado de 51 casos a cada 100 mil mulheres. A estratégia de tratamento das pacientes com tumores de mama pode passar pelo uso de quimioterapia. A doxorrubicina é uma das drogas mais ativas para o câncer de mama, pertencendo ao grupo das antraciclinas. A família das antraciclinas apresenta como efeito colateral dano ao miocárdio que pode chegar a 36% dependendo da dose utilizada. O efeito sobre o miocárdio costuma ocorrer mais comumente durante ou logo após o último ciclo de quimioterapia podendo, entretanto ocorrer após vários anos do último ciclo de quimioterapia. O objetivo deste estudo foi analisar as alterações da função diastólica ventricular esquerda em mulheres usuárias de antraciclínicos no tratamento do câncer de mama. Realizamos um estudo prospectivo, em uma coorte de mulheres entre 18 e 69 anos, com câncer de mama e indicação de quimioterapia com doxorrubicina. Acompanhamos por período não inferior a 18 meses um grupo de 38 pacientes que cumpriram os critérios de elegibilidade. A dose de doxorrubicina utilizada variou de 50 a 60 mg/m/SC. Todos os pacientes são do sexo feminino, e portadores do tipo histológico carcinoma ductal infiltrante. Duas pacientes faleceram durante o estudo, de causa não cardíaca. Em nossa avaliação, ao final do estudo observamos que os parâmetros: dimensões do átrio esquerdo, dimensões do ventrículo esquerdo na diástole, dimensões do ventrículo esquerdo na sístole, velocidade da onda E, relação da fase de enchimento rápido pela sístole atrial, velocidade diastólica tardia do anel mitral, velocidade diastólica precoce do anel mitral, tempo de desaceleração e a relação da velocidade de enchimento rápido precoce de VE pela velocidade diastólica precoce do anel mitral demonstraram serem parâmetros de grande utilidade para seguimento da lesão cardíaca por antraciclínicos. Já o que não ocorreu com: a fração de encurtamento, fração de ejeção, volume do AE, volume do AE corrigido pela superfície corporal, velocidade diastólica tardia, tempo de relaxamento isovolumétrico, velocidade sistólica do anel mitral, que não apresentaram alterações significativas neste estudo. A análise da função diastólica utilizando o ecocardiograma mostrou ser um método eficaz, que em conjunto com a da função sistólica possibilita detectar precocemente o possível dano miocárdico, oriundo ao uso da quimioterapia com antraciclínicos, favorecendo uma intervenção terapêutica precoce e adequada. / Breast cancer is one of the most important public health of our country. It was estimated for 2010, 49,400 new cases of breast cancer in Brazil, with an estimated risk of 51 cases per 100 000 women. The treatment strategy of patients with breast tumors can pass through the use of chemotherapy. Doxorubicin is one of the most active drugs for breast cancer, belonging to the group of anthracyclines. The family of anthracyclines has as side effect myocardial damage that can reach 36% depending on the dose used. The effect on the myocardium is known to occur most commonly during or shortly after the last cycle of chemotherapy, but it could occur several years after the last cycle of chemotherapy. The objective of this study was to evaluate changes in left ventricular diastolic function in women users of anthracyclines in the treatment of breast cancer. A prospective study in a cohort of women aged 18 to 69 years, with breast cancer and indications for chemotherapy with doxorubicin. We follow a period of not less than 18 months a group of 38 patients who met the eligibility criteria. The doxorubicin dose used ranged from 50 to 60 mg / m / SC. All patients were female, and carriers of infiltrating ductal carcinoma histology. Two patients died during the study of noncardiac causes. In our view, the end of the study showed that the parameters: size of left atrium (LA), left ventricular (LV) dimensions in diastole, left ventricular dimensions in systole, E wave velocity, ratio of phase rapid filling by atrial systole, late diastolic velocity of mitral annulus, early diastolic velocity of mitral annulus, deceleration time and speed ratio early rapid filling of the LV early diastolic velocity of mitral annulus proved to be useful parameters for monitoring of cardiac injury by anthracyclines. Although this did not occur with: fractional shortening, ejection fraction, LA volume, LA volume corrected for body surface area, late diastolic velocity, isovolumetric relaxation time, mitral annular systolic velocity, which showed no significant changes in this study. The analysis of diastolic function using echocardiography proved to be an effective method, which, together with the systolic function allows early detection of possible myocardial damage, arising the use of chemotherapy with anthracycline, favoring an early and appropriate therapeutic intervention.

O Câncer de mama é um dos problemas de saúde pública mais importantes em nosso país. São estimados, para 2010, 49.400 novos casos de câncer de mama no Brasil, com um risco estimado de 51 casos a cada 100 mil mulheres. A estratégia de tratamento das pacientes com tumores de mama pode passar pelo uso de quimioterapia. A doxorrubicina é uma das drogas mais ativas para o câncer de mama, pertencendo ao grupo das antraciclinas. A família das antraciclinas apresenta como efeito colateral dano ao miocárdio que pode chegar a 36% dependendo da dose utilizada. O efeito sobre o miocárdio costuma ocorrer mais comumente durante ou logo após o último ciclo de quimioterapia podendo, entretanto ocorrer após vários anos do último ciclo de quimioterapia. O objetivo deste estudo foi analisar as alterações da função diastólica ventricular esquerda em mulheres usuárias de antraciclínicos no tratamento do câncer de mama. Realizamos um estudo prospectivo, em uma coorte de mulheres entre 18 e 69 anos, com câncer de mama e indicação de quimioterapia com doxorrubicina. Acompanhamos por período não inferior a 18 meses um grupo de 38 pacientes que cumpriram os critérios de elegibilidade. A dose de doxorrubicina utilizada variou de 50 a 60 mg/m/SC. Todos os pacientes são do sexo feminino, e portadores do tipo histológico carcinoma ductal infiltrante. Duas pacientes faleceram durante o estudo, de causa não cardíaca. Em nossa avaliação, ao final do estudo observamos que os parâmetros: dimensões do átrio esquerdo, dimensões do ventrículo esquerdo na diástole, dimensões do ventrículo esquerdo na sístole, velocidade da onda E, relação da fase de enchimento rápido pela sístole atrial, velocidade diastólica tardia do anel mitral, velocidade diastólica precoce do anel mitral, tempo de desaceleração e a relação da velocidade de enchimento rápido precoce de VE pela velocidade diastólica precoce do anel mitral demonstraram serem parâmetros de grande utilidade para seguimento da lesão cardíaca por antraciclínicos. Já o que não ocorreu com: a fração de encurtamento, fração de ejeção, volume do AE, volume do AE corrigido pela superfície corporal, velocidade diastólica tardia, tempo de relaxamento isovolumétrico, velocidade sistólica do anel mitral, que não apresentaram alterações significativas neste estudo. A análise da função diastólica utilizando o ecocardiograma mostrou ser um método eficaz, que em conjunto com a da função sistólica possibilita detectar precocemente o possível dano miocárdico, oriundo ao uso da quimioterapia com antraciclínicos, favorecendo uma intervenção terapêutica precoce e adequada. / Breast cancer is one of the most important public health of our country. It was estimated for 2010, 49,400 new cases of breast cancer in Brazil, with an estimated risk of 51 cases per 100 000 women. The treatment strategy of patients with breast tumors can pass through the use of chemotherapy. Doxorubicin is one of the most active drugs for breast cancer, belonging to the group of anthracyclines. The family of anthracyclines has as side effect myocardial damage that can reach 36% depending on the dose used. The effect on the myocardium is known to occur most commonly during or shortly after the last cycle of chemotherapy, but it could occur several years after the last cycle of chemotherapy. The objective of this study was to evaluate changes in left ventricular diastolic function in women users of anthracyclines in the treatment of breast cancer. A prospective study in a cohort of women aged 18 to 69 years, with breast cancer and indications for chemotherapy with doxorubicin. We follow a period of not less than 18 months a group of 38 patients who met the eligibility criteria. The doxorubicin dose used ranged from 50 to 60 mg / m / SC. All patients were female, and carriers of infiltrating ductal carcinoma histology. Two patients died during the study of noncardiac causes. In our view, the end of the study showed that the parameters: size of left atrium (LA), left ventricular (LV) dimensions in diastole, left ventricular dimensions in systole, E wave velocity, ratio of phase rapid filling by atrial systole, late diastolic velocity of mitral annulus, early diastolic velocity of mitral annulus, deceleration time and speed ratio early rapid filling of the LV early diastolic velocity of mitral annulus proved to be useful parameters for monitoring of cardiac injury by anthracyclines. Although this did not occur with: fractional shortening, ejection fraction, LA volume, LA volume corrected for body surface area, late diastolic velocity, isovolumetric relaxation time, mitral annular systolic velocity, which showed no significant changes in this study. The analysis of diastolic function using echocardiography proved to be an effective method, which, together with the systolic function allows early detection of possible myocardial damage, arising the use of chemotherapy with anthracycline, favoring an early and appropriate therapeutic intervention.

Notre deuxième travail expérimental visait à élucider le rôle de la surcharge pondérale dans le développement de la cardiotoxicité des anthracyclines et du trastuzumab. Grâce à un modèle murin de surpoids modéré et de risque cardio-métabolique accru induits par programmation post-natale, nous avons mis en évidence le rôle potentiateur d’une surcharge pondérale sur le développement de la cardiotoxicité aux anthracyclines ; alors que la cardiotoxicité du trastuzumab ne semble pas être en revanche majorée par le surpoids. Nos travaux ont également permis de préciser les conditions dans lesquelles existent des potentialisations des effets lors de l’association doxorubicine et trastuzumab. / Cancer treatment has advanced considerably in recent years, allowing a reduction in mortality. Longer life expectancy of patients has helped to highlight the delayed onset of cardiovascular toxicity induced by these chemotherapies. The pathophysiological mechanisms responsible for these cardiac dysfunctions are complex, entangled and remain partially unknown. A better understanding of the phenomena involved in these cardiotoxicities is needed to prevent their occurrence. Therefore, we have developed two different experimental approaches to understand the pathophysiological mechanisms involved in the cardiac toxicity of anthracyclines and trastuzumab.A first experimental study aimed to clarify the role of iron in heart failure induced by anthracyclines. We have demonstrated that a tissular iron overload in mice prior to doxorubicin injection does not increase the cardiotoxicity of chemotherapy. On the contrary, the involvement of anti-radical defenses following the iron load could reduce cardiac oxidative damage generated by doxorubicin. In view of these data, the role of iron chelators in cardioprotection against anthracyclines has to be questioned.Our second experimental work was to elucidate the role of overweight in the development of anthracycline and trastuzumab cardiotoxicity. Using a mouse model of moderate overweight and of increased risk of cardiometabolic induced postnatal programming, we have highlighted the role of overweight on the development of anthracycline cardiotoxicity; whereas trastuzumab cardiotoxicity did not appear to be increased by overweight. Our work also clarified the conditions in which there are cumulative cardiac alterations when doxorubicin and trastuzumab are associated.

INTRODUÇÃO: Cardiotoxicidade é frequentemente associada ao uso crônico de doxorubicina (DOX) podendo levar a cardiomiopatia e insuficiência cardíaca. A identificação de miRNAs cardiotoxicidade-específicos e seu potencial como biomarcadores poderia fornecer uma ferramenta prognostica valiosa e uma potencial área de intervenção. METODOLOGIA: Este é um sub-estudo do ensaio clínico prospectivo \"Efeito do Carvedilol na Prevenção da Cardiotoxicidade Induzida por Quimioterapia\" (ensaio CECCY) no qual incluiu 56 pacientes do sexo feminino (idade 49.9±3.3) provenientes do braço placebo. Os pacientes incluídos foram submetidos à quimioterapia com DOX seguido por taxanos. Troponina cardiaca I (cTnI), fração de ejeção do ventrículo esquerdo (FEVE) e microRNAs foram mensurados periodicamente. RESULTADOS: Os níveis circulantes de miR-1, -133b, -146a e -423-5p aumentaram significativamente durante o tratamento (18.6, 11.5, 10.6 e 12.1-vezes respectivamente; p < 0.001) enquanto miR-208a e -208b foram indetectáveis. cTnI aumentou de 6.6 ± 0.3 para 46.7 ± 5.5 pg/ml (p < 0.001) enquanto FEVE tendeu a diminuir de 65.3±0.5 para 63.8±0.9 (p=0.053) após 12 meses; deis pacientes (17.9%) desenvolveram cardiotoxicidade. miR-1 foi associado a mudanças na FEVE (r2=0.363, p < 0.001) enquanto miR-1 e -133b foram associados a cTnI (r2 = 0.675 e 0.758; p < 0.001). Além disso, miR-1 antecipou a cardiotoxicidade e mostrou uma area sobre a curva maior que cTnI para discriminar pacientes que desenvolveram cardiotoxicidade daqueles que não desenvolveram (AUC = 0.849 e 456, p<0.001 e 0.663, respectivamente). CONCLUSÃO: Nossos dados sugerem miR-1 como um potencial novo biomarcador de cardiotoxicidade induzida por DOX em pacientes com câncer de mama. Estes resultados podem levar a novas estratégias de detecção precoce do risco de lesão cardíaca induzida por DOX bem como a introdução de uma nova área para intervenção / INTRODUCTION: Cardiotoxicity is frequently associated with the chronic use of doxorubicin (DOX) and may lead to cardiomyopathy and heart failure. Identification of cardiotoxicity-specific miRNA biomarkers could provide clinicians with a valuable prognosis tool and a potential area for intervention. METHODS: This is an ancillary study from the prospective trial \"Carvedilol Effect in Preventing Chemotherapy-Induced Cardiotoxicity.\" (CECCY trial) which included 56 female patients (49.9±3.3 age) from placebo arm. Enrolled patients were treated with DOX followed by taxanes. Cardiac troponin I (cTnI), left ventricle ejection fraction (LVEF) and miRNAs were measured periodically. RESULTS: Circulating levels of miR-1, -133b, -146a and -423-5p increased along the treatment (18.6, 11.5, 10.6 and 12.1-fold respectively; p < 0.001); miR-208a and -208b were undetectable. cTnI increased from 6.6±0.3 to 46.7 ± 5.5 pg/ml (p < 0.001) while LVEF tended to decrease from 65.3±0.5 to 63.8±0.9 (p=0.053) over 12 months; ten patients (17.9%) developed cardiotoxicity. miR-1 was associated to changes in LVEF (r2=0.363, p < 0.001) while miR-1 and -133b were associated to cTnI (r2 = 0.675 and 0.758; p < 0.001). Furthermore, miR-1 anticipated cardiotoxicity and showed greater area under the curve than cTnI to discriminate between patients who did and did not developed cardiotoxicity (AUC = 0.849 and 456, p < 0.001 and 0.663, respectively). CONCLUSION: Our data suggest circulating miR-1 as a potential new biomarker of DOX-induced cardiotoxicity in breast cancer patients. These results may lead to new earlier strategies to detect drug-induced cardiac injury risk before it develops to an irreversible stage or introduce new area for intervention

This dissertation reports the preparation of several types of anthraquinones structurally related to adriamycin. It describes the synthesis of two types of 2-aminoquinizarin compounds. It also presents two new syntheses of a heterocyclic tetracyclic ring system, similar to the aglicone ring system of adriamycin. A series of 2-aminoquinizarins was prepared by adding several primary amines to quinizarin. Quinizarin was shown to be essentially inert toward secondary amines. Several secondary amine adducts with quinizarin have been prepared, however, by treating the bis-boroacetate ester of quinizarin with the amines. Both types of 2-aminoquinizarin compounds exhibit outstanding potential for possessing antineoplastic activity, and several have been submitted to the National Cancer Institute for testing in their screening program for antineoplastic agents.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lucie Čermáková Supervisor: RNDr. Eva Novotná, Ph.D. Title of diploma thesis: The effect of acalabrutinib and ibrutinib on the efficacy of daunorubicin in cancer cells Leukemia presents malignant diseases of hematopoiesis, which essence is the malignant transformation of a hematopoietic stem cell at various levels of maturation and increased proliferative activity. Chemotherapy is the gold standard in the treatment of leukemia. One of the many treatments is the use of anthracycline chemotherapeutics, especially daunorubicin (DAU). Anthracyclines are widely used in clinical practice but have high cardiotoxic effects that limit their dosage. One of the main causes of side effects is the reduction of an anthracycline chemotherapeutic to the appropriate toxic metabolite, which accumulates in the heart. Carbonyl, reducing enzymes from the superfamily aldo-ketoreductase (AKR), and short-chain dehydrogenase/reductase (SDR) are involved in this reduction. At the same time, carbonyl reducing enzymes, has been shown to be involved in the mechanisms that cause tumor cells to be resistant to anthracyclines, thereby reducing the inhibition of the growth of these cells. In the diploma thesis we found that...