Sinteza fluoroforno označenih zaviralcev topoizomeraze IIα

Kopčavar, Zala (author), Ilaš, Janez (mentor)

Rakava obolenja so eden izmed najpogostejših vzrokov smrti na svetu, saj so zapletena in obremenjujoča stanja s kompleksnim mehanizmom nastanka. Kljub številnim novoodkritim kemoterapevtikom ostajajo nezadostna selektivnost, resni neželeni učinki in razvoj odpornosti na terapijo ključne pomanjkljivosti. Zaradi citotoksičnosti in razvoja odpornosti proti že uveljavljenim učinkovinam nenehno potekajo številne raziskave za odkrivanje novih učinkovin proti raku. Encimi topoizomeraze imajo pomembno vlogo v celičnih procesih in so bistveni za preživetje, zato njihova inhibicija vodi v celično smrt. Spreminjajo topologijo DNA z ustvarjanjem eno- ali dvoverižnih prehodnih prelomov v dvojni vijačnici DNA. Na podlagi strukture in mehanizma delovanja jih delimo na topoizomeraze tipa I in tipa II. Topoizomeraza IIα je znana in validirana tarča mnogih kemoterapevtskih zdravil. Pomen topoizomeraze IIα v delečih se celicah nakazuje, da je katalitična inhibicija encima lahko koristna strategija proti raku. Mnoga mesta v katalitičnem ciklu topoizomeraze še niso dobro izkoriščena kot vezavna mesta učinkovin, zato ostaja veliko potenciala za razvoj novih, učinkovitejših in pacientu prijaznejših zdravil. Kumarini imajo, kot fluorescenčne organske molekule, visoko biološko aktivnost, nizko citotoksičnost in izkazujejo protitumorno delovanje v več fazah nastanka raka preko različnih mehanizmov, vključno z zaviranjem encimov, kot je topoizomeraza. Eksperimentalni del magistrske naloge je osredotočen na oblikovanje novih fluoroforno označenih katalitičnih zaviralcev topoizomeraze IIα - učinkovin, ki tekmujejo z ATP za vezavno mesto na N-terminalnem delu encima in s tem ohromijo katalitični cikel encima. Tekom eksperimentalnega dela so bili sintetizirani in analizirani novi fluoroforno označeni katalitični zaviralci topoizomeraze IIα 3,4-dikloro-5-metilpirolamidnega in 5-klorotiofenskega tipa. Uporabljene sintezne poti so bile optimizirane, dobljeni produkti so bili zadovoljive kvantitete in čistosti. Končnim spojinam je bila z biokemijskim testiranjem določena encimska zaviralna aktivnost na DNA topoizomerazi IIα pri koncentracijah zaviralca 10 μM in 100 μM.

• Language: Slovenian
• Keywords: topoizomeraza IIα, ATP-kompetitivni zaviralci, fluoroforno označeni katalitični zaviralci topoizomeraze, kumarin (2H-kromen-2-on, 2H-1-benzopiran-2-on)
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-158514
• Publication date in RUL: 14.06.2024

Secondary language

• Language: English
• Title: Synthesis of fluorophore-labeled inhibitors of topoisomerase IIα

Abstract

Cancer is one of the world's leading causes of death, as they are complicated and burdensome conditions with complex mechanisms. Despite many newly discovered chemotherapeutics, insufficient selectivity, serious adverse events and development of resistance to therapy remain key drawbacks. Due to cytotoxicity and the development of resistance to established agents, constant research is ongoing to discover new anticancer agents. Topoisomerase enzymes play an important role in cellular processes and are essential for survival, so their inhibition leads to cell death. They alter the topology of DNA by creating single- or double-stranded transient breaks in the DNA double helix. Based on their structure and mechanism of action, they are divided into type I and type II topoisomerases. Topoisomerase IIα is a known and validated target of many chemotherapeutic drugs. The importance of topoisomerase IIα in proliferating cells suggests that catalytic inhibition of the enzyme may be a useful anticancer strategy. Many sites in the catalytic cycle of topoisomerase are not yet well exploited as binding sites for active substances, leaving much potential for the development of new, more effective and patient-friendly drugs. As fluorescent organic molecules, coumarins have high biological activity, low cytotoxicity and exhibit antitumour activity in several stages of cancer through various mechanisms, including inhibition of enzymes such as topoisomerase. The experimental part of this master’s thesis is focused on the design of novel fluorophore-labelled catalytic inhibitors of topoisomerase IIα - agents that compete with ATP for the binding site on the N-terminal part of the enzyme, thereby paralysing the catalytic cycle of the enzyme. In the course of the experimental work, new fluorophore-labelled catalytic inhibitors of topoisomerase IIα with 3,4-dichloro-5-methylpyrrolamide and 5-chlorothiophene scaffold were synthesised and analysed. The synthetic routes used were optimised and the products obtained were of satisfactory quantity and purity. The final compounds were evaluated by biochemical testing for enzyme inhibitory activity on DNA topoisomerase IIα at inhibitor concentrations of 10 μM and 100 μM.

• Keywords: topoisomerase IIα, ATP-competitive inhibitors, fluorophore-labeled catalytic topoisomerase inhibitors, coumarin (2H-chromen-2-one, 2H-1-benzopyran-2-one)