20.500.12556/RUL-129649 Sinteza spojine NSC94914 kot inhibitorja aktivnosti cisteinske peptidaze katepsina K Synthesis of compound NSC94914 as inhibitor of cysteine peptidase cathepsin K activity Katepsin K je cisteinska peptidaza, ki velja za tarčo pri zdravljenju osteoporoze. Eden izmed načinov za inhibicijo tega encima je ciljanje alosteričnih mest. Znanih je več alosteričnih inhibitorjev katepsina K. Pri sintezi enega od njih, spojine NSC94914 (2-([1,1'-bifenil]-2-ilmetil)malonske kisline), sem se najprej lotila nukleofilne substitucije. Sprva sem z NaH deprotonirala dibenzil malonat, katerega sem kasneje reagirala z 2-fenilbenzil bromidom. Dobljeni, s kolonsko kromatografijo očiščen, dibenzilni ester sem katalitsko hidrogenirala ter tako prišla do želenega produkta NSC94914. S pomočjo encimskih testov sem s fluorogenim substratom Z-FR-AMC okarakterizirala spojino NSC94914, določila način inhibicije ter afiniteto vezave. Izkazalo se je, da spojina povzroči popolno inhibicijo. Rezultat nasprotuje dosedanjim ugotovitvam raziskav, kar bi lahko bila posledica nečiste spojine NSC94914 ali izgubljene aktivnosti encima tekom testiranja. Cathepsin K is a cysteine peptidase, that is considered a target for the treatment of osteoporosis. One way to inhibit this enzyme is targeting allosteric sites. Several allosteric inhibitors of cathepsin K are known. In the synthesis of inhibitor NSC94914, 2-([1,1’-biphenyl]-2-ylmethyl)malonic acid, I firstly undertook the nucleophilic substitution. Initially, I deprotonated dibenzyl malonate with NaH and later reacted it with 2-phenylbenzyl bromide. After purification by column chromatography, the obtained dibenzyl ester was catalytically hydrogenated to get the desired product NSC94914. Using enzyme assays with the fluorogenic substrate Z-FR-AMC, I characterized NSC94914, determined the method of inhibition and the binding affinity. It has been shown, that this compound causes a complete inhibition. The result contradicts the published research findings, which could be due to the impure compound NSC94914 or lost enzyme activity during testing. katepsin K alosterična regulacija inhibicija NSC94914 cathepsin K allosteric regulation inhibition NSC94914 true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2021-09-06 12:41:04 2021-09-06 12:41:06 2022-09-08 04:09:51 0000-00-00 00:00:00 2021 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 18341 78798339 Bozic_Kristina_-_Sinteza_spojine_NSC94914_kot_inhibitorja_aktivnosti_cisteinske_peptidaze_kateps.pdf Bozic_Kristina_-_Sinteza_spojine_NSC94914_kot_inhibitorja_aktivnosti_cisteinske_peptidaze_kateps.pdf 1 8792438A93EFAA1121F82C56738678E2 3577fcb155ae8eef809be2d570ef6296de85b93b3039efa935c25ad04dcdb195 b26387ba-0efe-11ec-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=146665 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0