Sinteza fluorofornih zaviralcev topoizomeraze IIα

Tomaš, Rok (author), Ilaš, Janez (mentor)

Rakava obolenja ter samo zdravljenje v trenutnem času pomenijo veliko problematiko za zdravstveno stroko ter javno zdravstvo po vsem svetu. Gre za enega od vodilnih vzrokov smrti tako v razvitem svetu kot drugod. Samo zdravljenje te bolezni še vedno predstavlja velik problem v zdravstvu. Drugi problematični vidiki so tudi omejen nabor učinkovin, njihova učinkovitost ter selektivnost zoper rakave celice. Zaradi tega gre razvoj učinkovin v smeri odkrivanja tarč, ki bi ponujale bolj selektivno zdravljenje te bolezni skupaj z zmanjšano pojavnostjo stranskih učinkov terapije. Topoizomeraza tipa IIα je dobra tarča, saj je izražanje tega encima povečano v hitro deljivih celicah, kar pa je tudi lastnost rakavih celic. Sam encim katalizira precej kompleksen proces, zato je nabor vezavnih mest kar obsežen. Ena izmed možnosti je tudi vezavno mesto molekule ATP. Učinkovine, ki delujejo na to mesto, so katalitični zaviralci topoizomeraze in s svojim delovanjem ponujajo drugačen in bolj selektiven način zdravljenja od, sedaj že klinično uporabljenih, topoizomeraznih strupov. V tej magistrski nalogi se bomo osredotočili na sintezo novih katalitičnih zaviralcev človeške topoizomeraze IIα. Te učinkovine tekmujejo z molekulo ATP za vezavno mesto na N-terminalnemu delu encima, s tem pa preprečijo oz. zaustavijo katalitični cikel encima. Eksperimentalni del je obsegal sintezo in analizo novih katalitičnih zaviralcev topoizomeraze IIα 3,4-dikloro-5-metilpiroloamidnega tipa, ki delujejo kot kompetitivni zaviralci (tekmujejo z molekulo ATP za vezavno mesto). Kot dodatno pa smo v samo strukturo zaviralca vključili še fluoroforni skelet. Med sinteznim delom smo si pomagali z uporabo tekočinskih kromatografskih metod, masno spektroskopijo, sklopljeno s tekočinsko kromatografijo. Kot orodje za spremljanje reakcij smo uporabljali tankoplastno kromatografijo ter jedrsko magnetno resonanco za potrditev strukture spojin. Smiselno bi bilo ponoviti reakcijske korake za pridobitev končnih spojin z namenom vrednotenja zaviralnih sposobnosti, IC50 ter fluorofornih lastnosti. Rezultate bi lahko uporabili za načrtovanje novih derivatov ter izboljšanje topnosti samih učinkovin.

• Language: Slovenian
• Keywords: topoizomeraza IIα, ATP-kompetitivni zaviralci, protitumorne učinkovine, katalitični inhibitorji topoizomeraze, fluorofori, N-fenilpirolamidi
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-156435
• Publication date in RUL: 25.05.2024

Secondary language

• Language: English
• Title: Synthesis of fluorophoric topoisomerase IIα inhibitors

Abstract

Cancer diseases and the treatment itself, at the current time, represent a major problem for the medical profession and public health around the world. They are one of the leading causes of death both in the developed world and elsewhere. The treatment itself still represents a big problem, as does the range of active substances, their effectiveness and selectivity against cancer cells. For this reason, the development of active agents is directed towards the discovery of targets that would offer a more selective treatment of this disease together with a reduced incidence of side effects of the therapy. Type IIα topoisomerase represents a good potential target, as the expression of this enzyme is increased in rapidly dividing cells, which is also a characteristic of cancer cells. The enzyme itself catalyzes a fairly complex process, so the set of targets is quite extensive. One of the possibilities is the binding site of the ATP molecule. The active substances that act on this site are catalytic topoisomerase inhibitors and with their action offer a different and more selective method of treatment than the currently clinically used topoisomerase poisons. During this master's thesis, we will focus on the synthesis of new catalytic inhibitors of human topoisomerase IIα. These active substances compete with the ATP molecule for the binding site on the N-terminal part of the enzyme, thereby preventing or they stop the catalytic cycle of the enzyme. The experimental part consisted of the synthesis and analysis of new catalytic inhibitors of topoisomerase IIα of the 3,4-dichloro-5-methylpyrrolamide type, which act as competitive inhibitors (competing with the ATP molecule for the binding site). In addition, we included a fluorophore scaffolding in the structure of the inhibitor itself. During the synthesis work, we were aided by using liquid chromatography methods, mass spectroscopy coupled with liquid chromatography. We used thin-layer chromatography and nuclear magnetic resonance to confirm the structure of the compounds as a tool for monitoring the reactions. It would be reasonable to repeat the reaction steps to obtain the final compounds in order to evaluate the inhibitory abilities, IC50 and fluorophoric properties. The results could be used to design new derivatives and improve the solubility of the active ingredients themselves.

• Keywords: topoisomerase IIα, ATP-competitive inhibitors, antitumor agents, catalytic topoisomerase inhibitors, fluorophores, N-phenylpyrrolamides