20.500.12556/RUL-181503 Vpliv substituentov na karbamatni skupini na jakost kovalentnih zaviralcev butirilholin esteraze Influence of substituents on carbamate moiety on the potency of covalent butyrylcholine esterase inhibitors Alzheimerjeva bolezen je kompleksna, napredujoča nevrodegenerativna motnja s številnimi patološkimi spremembami centralnega živčnega sistema, ki se kažejo kot holinergična disfunkcija, kopičenje plakov in oligomerov amiloida beta, hiperfosforilacija proteina tau in nevrovnetje. Pomembna tarča v razvoju novih zdravilnih učinkovin je encim butirilholin esteraza (BChE), saj ima pomembno vlogo pri slabšanju kognitivnih funkcij pacienta zaradi vpliva na prenos signalov prek holinergičnega sistema. V magistrski nalogi smo obravnavali BChE kot terapevtsko tarčo in načrtovali, sintetizirali in vrednotili pet novih selektivnih kovalentnih zaviralcev encima. Pri tem smo izhajali iz spojine DP-802. Sečninsko skupino omenjene spojine smo zamenjali s karbamatno, da bi prek te zamenjave dosegli kovalentno vezavo na Ser198 BChE. Dodatno smo na benzenovem obroču acetamidno skupino zamenjali z nitrilom, s čimer smo izboljšali topnost intermediatov v organskih topilih, uporabljenih tekom sinteze, nismo pa poslabšali topnosti končnih spojin v pufru. Na karbamatno skupino smo pripenjali različne substituente, da smo lahko vrednotili, kateri pripomorejo h kovalentni vezavi na tarčo. Z izbiro ustreznih ligandov smo želeli tudi povečati selektivnost na BChE napram acetilholin esterazi (AChE). Sintetizirali in izolirali smo pet končnih spojin 8a-8e, ki smo jih vrednotili in vitro na humanih encimih (hAChE in hBChE) s pomočjo metode po Ellmanu. Najmočnejši zaviralec BChE je spojina 8d, pri kateri benzilni fragment v primerjavi z neposredno vezanim fenilnim fragmentom na karbamatni dušik izboljša zaviralno delovanje za približno dva velikostna razreda. Kljub temu testirane spojine niso bile selektivne na hBChE, saj je bila selektivnost spojin 8c in 8d v primerjavi s hAChE približno 10-kratna, potrebna pa je vsaj 1000-kratna razlika. Smo pa pri spojinah 8c in 8d dosegli psevdo-ireverzibilno zaviranje tarče. Alzheimer’s disease (AD) is a complex, progressive neurodegenerative disorder characterised by pathological changes in the central nervous system, including cholinergic dysfunction, accumulation of amyloid beta plaques and oligomers, hyperphosphorylation of the tau protein, and neuroinflammation. An important target in the development of new therapeutic agents targeting AD is the enzyme butyrylcholinesterase (BChE), as it plays a significant role in the deterioration of cognitive functions by influencing signal transmission within the cholinergic system. In this Master’s thesis, five new covalent inhibitors of BChE were designed, synthesized, and evaluated, based on the lead compound DP-802. In this compound, the urea group was replaced with a carbamate moiety to enable covalent binding to Ser198 of BChE. Additionally, the acetamide group on the benzene ring was replaced with a nitrile group, improving the solubility of synthetic intermediates in organic solvents used during synthetic process, without compromising the solubility of the final compounds in aqueous buffer. Various substituents were introduced on the carbamate moiety to evaluate their contribution to covalent binding to the target enzyme. Ligand design also focused on achieving improved selectivity for BChE over acetylcholinesterase (AChE). Five final compounds (8a-8e) were synthesized and isolated and tested in vitro against human acetylcholinesterase (hAChE) and butyrylcholinesterase (hBChE) using Ellman’s assay. Compound 8d was identified as the most potent BChE inhibitor, with a benzyl fragment providing approximately two orders of magnitude higher inhibitory activity compared to a directly attached phenyl fragment on the nitrogen atom. Nevertheless, the compounds did not exhibit pronounced selectivity for hBChE, as compounds 8c and 8d showed only about 10-fold selectivity over hAChE, whereas at least a 1000-fold difference would be required. However, compounds 8c and 8d demonstrated pseudo-irreversible inhibition of the target enzyme. Alzheimerjeva bolezen butirilholin esteraza karbamatna funkcionalna skupina kovalentni zaviralci Alzheimer’s disease butyrylcholinesterase carbamate functional group covalent inhibitors true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-04-09 08:45:06 2026-04-09 08:45:27 2026-04-10 04:38:55 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 124977 31902.pdf 31902.pdf 1 5F436286020ABBBB61502CE23B1A7274 7b28880eb962c2630523add53cddcfbbe7ff18c9d75c65defcb16cdc45177c77 7e825c13-33df-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=232015 Fakulteta za farmacijo 0 0 0