PROTAC molekule za ciljanu razgradnju kinaze glikogen sintaze 3

master's thesis

• Title (english): PROTACs for targeted degradation of glycogen synthase kinase 3
• Author: Karla Kostadinov
• Mentor: Kristina Pavić (mentor)
• Committee member: Kristina Pavić (predsjednik povjerenstva)
• Committee member: Marina Marinović (član povjerenstva)
• Committee member: Davor Šakić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medicinal chemistry); Zagreb
• Defense date and country: 2025-04-17, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Ciljana razgradnja proteina (TPD) predstavlja inovativan pristup u istraživanju i razvoju lijekova koji koristi prirodne stanične mehanizme za selektivno uklanjanje patoloških proteina. Posebno je značajna za ciljanje proteina na koje se klasičnim lijekovima malim molekulama ne može učinkovito djelovati. TPD ima velik potencijal u liječenju karcinoma, autoimunih i neurodegenerativnih bolesti.
Jedna od ključnih skupina molekula u TPD-u su kimere koje usmjeravaju ciljanu proteolizu, (engl. Proteolysis Targeting Chimeras, PROTAC-i). Ovaj diplomski rad opisuje sintezu i karakterizaciju PROTAC-a koji ciljaju enzim kinazu glikogen sintaze 3 (GSK3), ključnu kinazu u Alzheimerovoj bolesti.
Opisani PROTAC-i se sastoje od liganda za GSK3 (harmina), liganda za cereblon E3 ligazu (talidomida) i alkiltriazolske poveznice s tri, odnosno četiri ugljikova atoma u alkilnom lancu. U svrhu priprave PROTAC-a bilo je potrebno sintetizirati odgovarajuće prekursore: alkin harmina 2 i azide 3a-b. Alkin 2 dobiven je alkiliranjem harmina propargil-bromidom, dok su azidi 3a-b dobiveni diazo prijenosom iz odgovarajućih N-tert-butiloksikarbonil-monozaštićenih diamina i diazo donora 1H-imidazol-1-sulfonil azid hidroklorida 1. Alkin 2 i azidi 3a-b povezani su preko 1,2,3-triazolskog prstena u jednom reakcijskom koraku Huisgenovom 1,3-dipolarnom cikloadicijom, a nakon uklanjanja zaštitne tert-butiloksikarbonilne (Boc) iz spojeva 4a-b, dobiveni su amini 5a-b. Reakcijom kondenzacije amina 5a-b s talidomidskom kiselinom uz kondenzacijski reagens HATU sintetizirani su PROTAC-i 6a i 6b.
Spojevi su karakterizirani pomoću ¹H i ¹³C NMR, MS i FT-IR spektroskopije. Prema SwissADME analizi, spojevi imaju ograničenu oralnu bioraspoloživost, ne prelaze krvno-moždanu barijeru, djeluju kao inhibitori enzima citokrom P450 te su supstrati P-glikoproteina. Njihova biološka aktivnost bit će ispitana u daljnjim istraživanjima koja prelaze okvire ovog diplomskog rada.

Abstract (english)

Targeted protein degradation (TPD) is an innovative approach in drug discovery and development that harnesses natural cellular mechanisms to selectively eliminate pathological proteins. It is especially important for targeting proteins that cannot be effectively modulated by conventional small-molecule drugs. TPD holds great potential in the treatment of cancer, autoimmune, and neurodegenerative diseases.
One of the major classes of molecules in TPD are proteolysis-targeting chimera (PROTAC). This master’s thesis focuses on the synthesis and characterization of PROTACs targeting glycogen synthase kinase 3 (GSK3), a key kinase implicated in Alzheimer’s disease.
The designed PROTACs consist of a GSK3 ligand (harmine), an E3 ligase ligand (thalidomide), and an alkyltriazole linker containing three or four carbon atoms in the alkyl chain. For the preparation of the target PROTACs, the appropriate precursors were synthesized: harmine alkyne 2 and azides 3a-b. Alkyne 2 was obtained by alkylating harmine with propargyl bromide, while azides 3a-b were synthesized by diazo transfer from the corresponding N-tert-butyloxycarbonyl-monoprotected diamines and the diazo donor 1H-imidazole-1-sulfonyl azide hydrochloride 1. Alkyne 2 and azides 3a-b were connected through a 1,2,3-triazole ring in a single reaction step using Huisgen 1,3-dipolar cycloaddition. After removal of the tert-butyloxycarbonyl (Boc) protecting group from compounds 4a-b, the corresponding amines 5a-b were obtained. Condensation of amines 5a-b with thalidomide acid using the coupling reagent HATU afforded PROTACs 6a and 6b.
The compounds were characterized using ¹H and ¹³C NMR, MS, and FT-IR spectroscopy. According to SwissADME predictions, the compounds exhibit limited oral bioavailability, do not cross the blood-brain barrier, act as cytochrome P450 inhibitors, and are substrates of P-glycoprotein. Their biological activity will be investigated in future studies beyond the scope of this thesis.

• Keywords: ciljana razgradnja proteina; PROTAC; GSK3; Alzheimerova bolest
• Keywords (english): targeted protein degradation; PROTAC; glycogen synthase kinase 3; Alzheimer’s disease
• Language: croatian
• URN:NBN: urn:nbn:hr:163:493971
• Study programme: Title: Pharmacy - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar farmacije (sveučilišni magistar farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2027-04-17
• Created on: 2025-04-24 10:33:16