Sinteza novih selektivnih zaviralcev topoizomeraze IIα s fenolnim ogrodjem

Lukić, David (author), Ilaš, Janez (mentor)

Rakave bolezni so velik javnozdravstveni problem, saj so glavni razlog za smrti širom celega sveta. Pri njihovi terapiji se srečujemo z veliko težavami, kot so nezadostna selektivnost učinkovin do tarčnih makromolekul, kot tudi razvoj rezistence na že obstoječe učinkovine in načine terapije. Pojavlja se potreba po bolj selektivnih učinkovinah in tudi tarčah z bolj specifičnim delovanjem, ciljano predvsem na tumorske celice. Topoizomeraza IIα je encim, ki se izraža predvsem v hitro delečih celicah in s tem omogoča usmerjeno zdravljenje, katerega tarča so predvsem tumorske celice. Zaradi kompleksnega katalitičnega cikla topoizomeraze IIα imamo na voljo več možnih tarč za delovanje učinkovin. V klinični praksi se že uporablja en razred učinkovin, ki zavirajo delovanje encima (topoizomerazni strupi), a je zaradi nekaterih resnih neželenih učinkov, kot tudi pojava rezistence na učinkovine, potreba po učinkovinah z novim mehanizmom delovanja. Zato se bomo v okviru eksperimentalnega dela te magistrske naloge osredotočili v oblikovanje novih katalitičnih zaviralcev topoizomeraze IIα - učinkovin, ki tekmujejo z ATP za vezavno mesto na N-terminalnem delu encima in s tem preprečijo delovanje katalitičnega cikla encima. Tekom eksperimentalnega dela smo sintetizirali in analizirali nove katalitične zaviralce topoizomeraze IIα 3,4-dikloro-5-metilpirolamidnega tipa, ki delujejo po principu tekmovanja z ATP za vezavno mesto. Za spremljanje sintezne poti smo uporabljali tekočinsko kromatografijo, sklopljeno z masno spektrometrijo, za potrjevanje struktur produktov smo uporabljali jedrsko magnetno resonanco in za sprotno spremljanje poteka reakcij smo uporabljali tankoplastno kromatografijo. Uporabljene sintezne poti so bile uspešne, saj smo v vseh primerih dobili produkte zadostne kvantitete in čistosti. Uporabili smo različne že znane metode izolacije in čiščenja produktov, ki so se izkazale kot robustne in učinkovite. V prihodnosti bi bilo smiselno ovrednotiti zaviralno aktivnost sintetiziranih spojin na topoizomerazi IIα in ovrednotiti fizikalno-kemijske lastnosti učinkovin. Na podlagi teh rezultatov bi se lahko pridobilo ključne podatke za nadaljnji razvoj in optimizacijo spojin.

• Language: Slovenian
• Keywords: topoizomeraza IIα, ATP-kompetitivni inhibitorji, učinkovine proti raku, katalitični inhibitorji topoizomeraze, N-fenilpirolamidi
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2022
• PID: 20.500.12556/RUL-143377
• Publication date in RUL: 17.12.2022

Secondary language

• Language: English
• Title: Synthesis of new selective topoisomerase IIα inhibitors with phenolic scaffold

Abstract

Cancer spectrum of diseases are the greatest cause of death worldwide, which is major public health concern. During treatment, we’re faced with many challenges, such as insufficient target receptor selectivity of compounds, development of resistance against compounds and treatments used not being effective anymore. There’s a need for more selective compounds with more specific function, targeted mainly at tumor cells. Topoisomerase IIα is an enzyme, which is present predominantly in fast-multiplying cells, allowing for more targeted treatment, aimed at mainly tumor cells. Because of complex catalytic cycle of topoisomerase IIα, we have several possible targets for compounds to act. In clinical practice there’s one class of compounds already in use - topoisomerase poisons, but because of some serious side effects as well as development of resistance to some compounds, there’s a need for compounds with different mechanism of action. Therefore, as a part of the experimental part of this master’s thesis, we’ll focus on the design of new catalytic inhibitors of topoisomerase IIα - compounds competing with ATP for the binding site on the N-terminal part of the enzyme and thus preventing the catalytic cycle from functioning. During experimental work we synthesized and analyzed new catalytic inhibitors of topoisomerase IIα of 3,4-dichloro-5-methylpyrrolamide type, which compete with ATP for the binding site. For monitoring the synthesis route, we used liquid chromatography coupled with mass spectrometry, nuclear magnetic resonance to confirm structure of compounds and thin-layer chromatography to monitor the progress of the reactions. The synthesis routes used were successful, since in all cases we obtained products of sufficient quantity and purity. We used various already known methods of product isolation and purification, which proved to be robust and effective. In the future, it would be sensible to evaluate the inhibitory activity of the synthesized compounds on topoisomerase IIα and to evaluate the physicochemical properties of the compounds. Based on these results, key data could be obtained for further development and optimization of compounds.

• Keywords: topoisomerase IIα, ATP-competitive inhibitors, anticancer compounds, catalytic inhibitors od topoisomerase, N-phenylpyrrolamides