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Expression of ABC transporters in cell cultures of non-small cell lung cancer: effect of selected targeted drugs
Žofajová, Barbora ; Hofman, Jakub (advisor) ; Červený, Lukáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Barbora Žofajová Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Expression of ABC transporters in non-small cell lung carcinoma cell cultures: effect of selected targeted drugs ABC transporters form a special family of transmembrane proteins, their function is the active efflux of substrates into the extracellular environment. These transporters are expressed in normal tissues and tumor cells. Thus, they influence drug pharmacokinetics and play a role in the development of multidrug resistance (MDR). Tyrosine kinases have essential functions ranging from proliferation to differentiation and cell death, their mutation can cause carcinogenesis. Finding, exploring, and inhibiting the functions of these targets is a promising strategy to treat cancer. This work aimed to determine whether the selected drugs are involved in pharmacokinetic interactions that are based on inductions of ABC efflux transporter genes and whether they have the potential to affect MDR phenotype of tumor cells. The drugs, which we studied, were three targeted anticancer drugs (capmatinib, pralsetinib, and tazemetostat) and two well- known chemotherapeutic drugs (pemetrexed and methotrexate). First, we...
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Study on the effect of selected targeted drugs on drug resistance mediated by ABC drug transporters
Paulusová, Viktória ; Hofman, Jakub (advisor) ; Novotná, Eva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Viktória Paulusová Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Title od diploma thesis: Study on the effect of selected targeted drugs on drug resistance mediated by ABC drug transporters. Multidrug resistance (MDR) is one of the major problems associated with cancer treatment. A key determinant of MDR is increased efflux of drugs through membrane ATP- binding cassette (ABC) transporters, which are a family of transporter proteins. Their overexpression contributes to MDR by decreasing intracellular drug concentration due to the efflux of various drugs from cells. Targeting ABC transporters appears to be a promising approach to suppress drug resistance. Therefore, one strategy to reverse the resistance of ABC transporter-expressing tumor cells is the combined use of chemotherapeutic drugs with ABC transporter modulators to enhance therapeutic efficacy. The aim of this work was to investigate the effect of targeted drugs (capmatinib, pralsetinib and tazemetostat) in combination with cytostatics (etoposide and topotecan) on drug resistance mediated by ABC efflux transporters. Results were obtained using MTT assays and caspase activity assays performed on the parental A431 cell line and its...
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The use of selected inhibitors to overcome anthracycline resistance in cancer therapy (in vitro study).
Tučková, Kateřina ; Wsól, Vladimír (advisor) ; Hofman, Jakub (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kateřina Tučková Supervisor: prof. Ing. Vladimír Wsól, Ph.D. Consultant: Mgr. Lenka Laštovičková, Ph.D. Title of diploma thesis: The use of selected inhibitors to overcome anthracycline resistance in cancer therapy (in vitro study) Resistance to chemotherapy is a severe problem in treating cancer patients, significantly reducing their chances of recovery. The increased expression of carbonyl reductases in tumour cells is one of the leading causes of the resistance. These enzymes can reduce anthracycline-based chemotherapy drugs to their less effective derivatives and thus significantly reduce their efficiency. Therefore, the inhibitors of carbonyl reductases could increase the effectiveness of anthracycline-based chemotherapy. The goal is to find an inhibitor with high inhibitory activity and low side effects. This thesis tested inhibitors asciminib, tucatinib, sotorasib, and umbralisib hydrochloride against carbonyl reductases from the aldo-keto reductase superfamily (AKR1C3, 1A1, 1B1, 1B10) and short-chain dehydrogenases/reductases superfamily (CBR1), using chemotherapy drug daunorubicin. The most significant inhibitory potential was observed between asciminib and the enzyme AKR1C3,...
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Study of the effect of ABC drug transporters' overexpression on the antiproliferative capacities of selected conventional cytostatics
Malečová, Lucie ; Hofman, Jakub (advisor) ; Trejtnar, František (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Malečová Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Study of the effect of ABC drug transporters' overexpression on the antiproliferative capacities of selected conventional cytostatics Overexpression of ABC drug transporters is one of the mechanisms of multiple drug resistance development, which results in therapy failure in cancer. ABC transporters such as ABCB1, ABCG2 and ABCC1 have a wide range of substrates, including several conventional cytostatics. This study aimed to examine the influence of overexpression of the mentioned transporters on the antiproliferation effects of docetaxel, etoposide, paclitaxel, methotrexate, pemetrexed, topotecan and vinblastin. The results were obtained with the MTT assay in which the MDCKII and A431 cell lines with/without overexpression of mentioned transporters were used. A decrease of cytotoxicity in the presence of at least one of the transporters was observed in five of seven tested cytostatics. Only topotecan has been proven to be the victim of overexpression of all of the tested transporters. To prove the applicability of the obtained results, topotecan was used in the follow-up combination study. Within this study, I have...
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Assessment of drug cytotoxicity using various cell viability tests
Kaňková, Kateřina ; Hofman, Jakub (advisor) ; Kozlíková, Magdaléna (referee)
V ANGLICKÉM JAZYCE Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Kateřina Kaňková Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Supervisor-consultant: doc. PharmDr. Ondřej Soukup, Ph.D. Title of diploma thesis: Assessment of drug cytotoxicity using various cell viability tests Nowadays, cell viability assays are an important part of preclinical studies. There is a wide spectrum of different methods that can be used for such purposes. This thesis aims to compare some of the fundamental methods, their accuracy and selectivity, as well as the advantages and drawbacks of their application. Six different drugs were measured using five different cell viability assays - colorimetric (MTT, crystal violet), fluorimetric (propidium iodide, calcein-AM) and luminometric (ATP) on the CHO-K1 cell line. The outcomes of all methods are IC50 values of used drugs assessing their cytotoxicity, including graphic representations of the measurements and standard deviations. Finally, all the methods are compared based on experience gained during the experiment as well as information gained during the theoretical research. Among the most suitable methods belong the MTT and ATP assay. Due to some difficulties during the measurement (e.g. unsuitable for use in...
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Study on interactions of selected targeted drugs with ABC drug transporters
Svobodová, Markéta ; Hofman, Jakub (advisor) ; Čečková, Martina (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Markéta Svobodová Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Title od diploma thesis: Study on interactions of selected targeted drugs with ABC drug transporters Cancer is one of the most common causes of death and its incidence is constantly increasing. Treatment mainly includes a combination of drug therapy, surgery and radiotherapy. A new approach, with huge potential, is the use of targeted anti-tumor therapy. This approach is based on specific interaction with tumor cells that reduces the negative effects occuring in conventional chemotherapy. Despite intensive research, we still face insufficient response to treatment. The main cause of therapy failure is the emergence of multidrug resistance (MDR) and the formation of metastases. The MDR phenomenon is most often associated with the overexpression of ATP binding cassette (ABC) transporters. These are responsible not only for the excessive efflux of anticancer drugs from cells, but also mainly affect their pharmacokinetics. Inhibition of these transporters increases the intracellular concentration of the cytostatic and provide the ability to modulate MDR. Therefore, one of the aims of this work was to investigate the inhibitory activity of...
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Assessment of drug cytotoxicity using various cell viability tests
Kaňková, Kateřina ; Hofman, Jakub (advisor) ; Kozlíková, Magdaléna (referee)
V ANGLICKÉM JAZYCE Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Kateřina Kaňková Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Supervisor-consultant: doc. PharmDr. Ondřej Soukup, Ph.D. Title of diploma thesis: Assessment of drug cytotoxicity using various cell viability tests Nowadays, cell viability assays are an important part of preclinical studies. There is a wide spectrum of different methods that can be used for such purposes. This thesis aims to compare some of the fundamental methods, their accuracy and selectivity, as well as the advantages and drawbacks of their application. Six different drugs were measured using five different cell viability assays - colorimetric (MTT, crystal violet), fluorimetric (propidium iodide, calcein-AM) and luminometric (ATP) on the CHO-K1 cell line. The outcomes of all methods are IC50 values of used drugs assessing their cytotoxicity, including graphic representations of the measurements and standard deviations. Finally, all the methods are compared based on experience gained during the experiment as well as information gained during the theoretical research. Among the most suitable methods belong the MTT and ATP assay. Due to some difficulties during the measurement (e.g. unsuitable for use in...
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Study of the effect of ABC drug transporters' overexpression on the antiproliferative capacities of selected conventional cytostatics
Malečová, Lucie ; Hofman, Jakub (advisor) ; Trejtnar, František (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Malečová Supervisor: doc. RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Study of the effect of ABC drug transporters' overexpression on the antiproliferative capacities of selected conventional cytostatics Overexpression of ABC drug transporters is one of the mechanisms of multiple drug resistance development, which results in therapy failure in cancer. ABC transporters such as ABCB1, ABCG2 and ABCC1 have a wide range of substrates, including several conventional cytostatics. This study aimed to examine the influence of overexpression of the mentioned transporters on the antiproliferation effects of docetaxel, etoposide, paclitaxel, methotrexate, pemetrexed, topotecan and vinblastin. The results were obtained with the MTT assay in which the MDCKII and A431 cell lines with/without overexpression of mentioned transporters were used. A decrease of cytotoxicity in the presence of at least one of the transporters was observed in five of seven tested cytostatics. Only topotecan has been proven to be the victim of overexpression of all of the tested transporters. To prove the applicability of the obtained results, topotecan was used in the follow-up combination study. Within this study, I have...
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Study of the role of nucleoside transporters in intestinal absorption with the use of in vitro transport method based on the Caco-2 cell line.
Ďurišová, Markéta ; Červený, Lukáš (advisor) ; Hofman, Jakub (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Mgr. Markéta Ďurišová Supervisor: PharmDr. Lukáš Červený, Ph.D. Title of rigorosum thesis: Study of the role of nucleoside transporters in intestinal absorption with the use of in vitro transport method based on the Caco-2 cell line. Perorally used drugs are primarly absorbed in the small intestine. Enterocytes, forming the intestinal barrier, express a large number of transporters, representing both ATP-binding (ABC) and "solute carrier" (SLC) transporters, which can significantly affect the pharmacokinetics of the drugs transmitted by them. For this reason, FDA and EMA are currently focusing on studying these interactions and their effects on drug permeability. The aim of this thesis was to investigate whether the equilibrative nucleoside transporters (ENTs), belonging to the SLC transporters, are involved in the absorption of nucleosides (adenosine and thymidine) and subsequently abacavir, the nucleoside antiretroviral. The in vitro transport method was performed on polarized, Caco-2 cell monolayer. By the statistical analysis of acquired data was found, that although the expression of ENTs was confirmed on the apical side of the Caco-2 cell membrane, the transport of nucleosides [3H]ADE and...
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Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method
Šinaľová, Lucia ; Hofman, Jakub (advisor) ; Jansová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucia Šinaľová Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method This master thesis focuses on the prediction of tacrine derivatives permeability through blood- brain barrier and their cytotoxicity. We studied tacrine-benzothiazole and tacrine- thiaquinazoline derivates as potential drugs for treatment of Alzheimer's disease. The reason of testing new tacrine derivates was effort to find substances with same effect and lower toxicity, for which was the tacrine discarded from clinical practice. The probable permeability of the studied substances was determined in vitro by the PAMPA method. Based on the permeation coeficient values we have identified that tacrine-benzothiazole derivates A-D (Pe= 8,31- 16,9×10-6 cm/s) and tacrine-thiaquinazoline derivates 1, 3, 4 (Pe= 8,59-14,9×10-6 cm/s) were permeable through blood-brain barrier, tacrine-thiaquinazoline derivate 2 (Pe= 3,79×10-6 cm/s) had uncertain permeability and tacrine-thiaquinazoline derivate 5 (Pe= 2,0×10-6 cm/s) was not permeable. In the pre-clinical evaluation of...
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