Sinteza in vrednotenje novih N-(2-benziloksi)-fenilpirolamidnih zaviralcev človeške DNA-topoizomeraze II

Handanagić, Bilka (author), Zidar, Nace (mentor)

Rak je kompleksna bolezen in en izmed glavnih vzrokov smrti po svetu. S staranjem prebivalstva in večanjem populacije bo imel vse večjo pojavnost ter zato predstavlja resen problem za javno zdravstvo in družbo nasploh. Da bi preprečili širjenje te bolezni, potekajo intenzivne raziskave odkrivanja selektivnih učinkovin za zdravljenje rakavih obolenj. Ena od glavnih lastnosti rakavih celic je, da se hitro in nenadzirano delijo. Encimi, ki sodelujejo pri podvajanju molekule DNA in so nujni za delitev celice ter za njeno preživetje, se imenujejo topoizomeraze. Zaradi pomembne vloge topoizomeraz pri celični delitvi smo človeško DNA topoizomerazo IIα, ki je bolj izražena v rakavih celicah, izkoristili kot tarčo za sintezo protirakavih zdravilnih učinkovin. Učinkovine, ki se trenutno uporabljajo za zdravljenje rakavih obolenj, imajo veliko neželenih učinkov, zato so nadaljnje raziskave nujne. Predmet magistrske naloge so spojine, ki se vežejo na ATP-vezavno mesto N terminalnega dela encima in predvidoma delujejo selektivno na človeško DNA topoizomerazo IIα. V okviru magistrske naloge smo sintetizirali in ovrednotili serijo novih N (2 benziloksi) fenilpirolamidnih ATP kompetitivnih zaviralcev človeške DNA topoizomeraze IIα. Pripravili smo tri končne spojine: 8, 9 in 14. Vse tri spojine smo najprej fizikalno-kemijsko ovrednotili in na ta način preverili ter potrdili njihovo identiteto in čistost. V nadaljnjem postopku smo vse tri spojine biološko ovrednotili, tako da smo določili rezidualno aktivnost (RA) človeške DNA topoizomeraze IIα pri koncentracijah spojin 40 µM in 4 µM. Spojini 8 in 14 nista izkazovali zaviralne aktivnosti. Zaradi njunih skupnih strukturnih lastnosti lahko sklepamo, da primarna amidna skupina ne tvori pomembnih interakcij z vezavnim mestom za ATP. Najboljšo zaviralno aktivnost je imela spojina 9, saj je pri njeni koncentraciji 40 µM RA encima znašala 60 %. Njeno večjo zaviralno aktivnost smo glede na strukturne elemente pripisali prisotnosti karboksilne skupine. Pridobljeni rezultati in ugotovitve magistrske naloge bodo koristili pri nadaljnjih raziskavah in pri razvoju boljših ATP-kompetitivnih zaviralcev DNA topoizomeraze IIα, ki bodo poleg dobrega zaviralnega delovanja imeli tudi sposobnost dostopa do tarče v celici.

• Language: Slovenian
• Keywords: rak, DNA topoizomeraza IIα, protirakava učinkovina, ATP-kompetitivni zaviralec, N (2 benziloksi) fenilpirolamid
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2020
• PID: 20.500.12556/RUL-117134
• Publication date in RUL: 25.06.2020

Secondary language

• Language: English
• Title: Synthesis and evaluation of novel N-(2-benzyloxy)-phenylpyrrolamides as inhibitors of human DNA topoisomerase II

Abstract

Cancer is a complex disease, one of the leading causes of death in the world, which will have an increasing incidence with the ageing of the population and its growth, posing a serious public health problem as well as a social problem. To prevent this, intensive research is underway to find a selective treatment for most cancers. One of the main characteristics of cancer cells is that they divide quickly and uncontrollably. The enzymes involved in the replication of a DNA molecule that are necessary for the cell division and its survival are called topoisomerases. Because of the important role of topoisomerases in cell division, human DNA topoisomerase IIα, which is highly expressed in cancer cells, has been selected as a target for the synthesis of anticancer active substances. Drugs that are currently used in cancer treatment have many side effects, which indicates a need for further research. The subject of this master's thesis is compounds that bind to the ATP-binding site of the N terminal portion of the enzyme and could presumably act selectively on human topoisomerase II. In the experimental part of the master's thesis, a series of new N (2 benzyloxy) phenylpyrrolamide ATP-competitive inhibitors of human DNA topoisomerase IIα were synthesized and evaluated. The synthesis yielded three final compounds, 8, 9, and 14. All three compounds were first physically and chemically evaluated to verify and confirm their identity and purity. In the following procedures, all three compounds were biologically evaluated by determining the residual activities (RAs) of human DNA topoisomerase IIα at 40 µM and 4 µM concentrations of tested substances. Compounds 8 and 14 did not show potent inhibitory activities. Considering their common structural properties, it can be concluded that the amide group does not form the necessary interactions with the ATP binding site. Compound 9 had the best inhibitory activity in the series, and at its 40 µM concentration, the RA of the enzyme was 60%. Its improved inhibitory activity can be attributed to the presence of a carboxylic acid group. The results and the findings of the master's thesis could be used in further research to obtain better optimized ATP-competitive inhibitors, which, in addition to excellent inhibitory ability, would also have the ability to reach the targets within cells.

• Keywords: cancer, DNA topoisomerase IIα, anticancer drug, ATP-competitive inhibitor, N (2 benzyloxy) phenylpyrrolamide