Sinteza spojine NSC94914 kot inhibitorja aktivnosti cisteinske peptidaze katepsina K

Božič, Kristina (avtor), Grošelj, Uroš (mentor)

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.

• Jezik: Slovenski jezik
• Ključne besede: katepsin K, alosterična regulacija, inhibicija, NSC94914
• Vrsta gradiva: Diplomsko delo/naloga
• Tipologija: 2.11 - Diplomsko delo
• Organizacija: FKKT - Fakulteta za kemijo in kemijsko tehnologijo
• Leto izida: 2021
• PID: 20.500.12556/RUL-129649
• COBISS.SI-ID: 78798339
• Datum objave v RUL: 06.09.2021

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Synthesis of compound NSC94914 as inhibitor of cysteine peptidase cathepsin K activity

Izvleček

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.

• Ključne besede: cathepsin K, allosteric regulation, inhibition, NSC94914