magistrski študij industrijska farmacija
Nataša Turk (Author), Nace Zidar (Mentor)

Abstract

Pomembno vlogo pri zdravljenju rakavih obolenj imajo zdravilne učinkovine, ki vplivajo na proces podvojevanja DNA. Mednje med drugim spadajo ATP-kompetitivni zaviralci DNA-topoizomeraze II, ki se vežejo na ATP-vezavno domeno N-terminalnega dela encima. S pomočjo informacij, ki smo jih dobili iz predhodno znanih kompleksov DNA-topoizomeraze II z vezanimi zaviralci smo sintetizirali serijo potencialnih zaviralcev tega encima, katerim je skupen N-(4-karbamoilfenil)pirolamidni del in so potencialni ATP kompetitivni zaviralci encima. Biološko smo ovrednotili štiri spojine (7, 8, 12 in 17), za katere smo, glede na njihovo zgradbo, predvidevali izrazito aktivnost. Spojinama 7 in 17 smo določili rezidualno aktivnost na DNA-topoizomerazi II pri 10 microM in 100 microM koncentraciji zaviralca, spojinama 8 in 12 pa smo določili tudi srednjo zaviralno koncentracijo (IC50) DNA-topoizomeraze II. Spojina 7 je bila z 91 rezidualno aktivnostjo pri 100 microM neaktivna, spojina 17 pa se je z 41% rezidualno aktivnostjo pri 100 microM izkazala kot šibek ATP-kompetitivni zaviralec. Spojini 8 in 12 sta imeli glede na etopozid (IC50 = 165 microM), ki je bil uporabljen kot pozitivna kontrola, precej večjo zaviralno aktivnost z nižjima vrednostma IC50 (2,97 microM in 3,59 microM). Spojinama 8 in 12 je skupna prosta karboksilna skupina, ki se je izkazala kot ključen strukturni del za zagotavljanje dobre zaviralne aktivnosti na encim. Spojina 7, ki namesto proste karboksilne skupine vsebuje metilno estrsko skupino, se je izkazala za neaktivno, prav tako spojina 17 z zaščitno skupino Boc. Spojine 8, 12 in 17 smo testirali tudi na celični liniji raka dojke. Spojini 8 in 12 sta bili neaktivni, za aktivno pa se je, prav tako v nasprotju z rezultati encimskih testiranj, izkazala spojina 17. Vzrok za nasprotujoče si rezultate encimskih in celičnih testiranj bi lahko pripisali prisotnosti karboksilne skupine, ki ji sicer pripisujemo pomembno vlogo pri vezavi na encim. N-(4-karbamoilfenil)pirolamidi so obetaven strukturni razred zaviralcev človeške DNA-topoizomeraze II. Pri nadaljnjem razvoju in raziskavah zaviralcev s podobno strukturo bi bilo smiselno raziskati vpliv prisotnosti karboksilne kisline na delovanje zaviralcev ter optimizirati sintezno pot za pripravo odščitenega analoga spojine 17.

Keywords

DNA-topoizomeraza II;protirakave učinkovine;ATP-kompetitivni zaviralci;N-(4-karbamoilfenil)pirolamidi;zaviralci angiogeneze;zaviralci sinteze DNA;sinteza spojin;sintezni postopki;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FFA - Faculty of Pharmacy
Publisher: [N. Turk]
UDC: 543.057:615:616-006(043.3)
COBISS: 4661617 Link will open in a new window
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Other data

Secondary language: English
Secondary title: Synthesis and evaluation of novel N-(4-carbamoylphenyl)pyrrolamides as human DNA topoisomerase II inhibitors
Secondary abstract: Drugs that target the DNA replication cycle have an important role in treatment of cancer. Among others, these include ATP-competitive inhibitors, which bind to the ATP binding domain on the N-terminal part of the enzyme. Having an understanding of the interactions between DNA-topoisomerase II and their inhibitors, a series of potential inhibitors was synthesized. They are all composed of N-(4-carbamoylphenyl)pyrrolamide structural core and are potential ATP competitive DNA topoisomerase II inhibitors. We biologically evaluated four compounds (7, 8, 12 in 17), for which we anticipated prominent activity according to their structure. Residual activities at inhibitor concentration of 10 [micro]M and 100 [micro]M were measured for compounds 7 and 17 and IC50 values were determined for compounds 8 and 12. Compound 7 with 91% residual activity at 100 %M is inactive, and compound 17 with 41% residual activity at 100 [micro]M is a weak ATP-competitor inhibitor. Compounds 8 and 12 have, relative to the positive control etoposide (IC50 = 165 [micro]M), a significantly higher inhibitory activity with lower IC50 values (2.97 [micro]M and 3.59 %M). Compounds 8 and 12 both possess a free carboxyl group, which has proven to be a key structural component to provide good inhibitory activity on the enzyme. Compound 7, which contains free methyl ester group instead of carboxyl group, as well as compound 17 containing the Boc protective group, were inactive. Compounds 8, 12, and 17 were also tested on cancer cell lines where compounds 8 and 12 proved to be inactive, and compound 17 proved active, all of which were not expected in regards to the enzyme test results. The cause of the contradictory results of the enzyme and cancer cell line testing could be attributed to the presence of carboxyl group, which is otherwise presumed to have an important role in binding to the enzyme. N-(4-carbamoylphenyl)pyrrolamides are a promising structural group of DNA topoisomerase II inhibitors. In the further development of similarly structured inhibitors it would be reasonable to investigate the effect of carboxylic acid presence on inhibitors' activity and to optimize the synthesis of Boc-deprotected analogue of compound.
Secondary keywords: Rak (medicina);
Type (COBISS): Master's thesis/paper
Thesis comment: Univ. Ljubljana, Fakulteta za farmacijo
Pages: VIII, 54 str.
ID: 11008000