Sinteza kovalentnih zaviralcev imunoproteasoma z amino-substituiranim piperidinskim obročem

Master's Study Programme Industrial Pharmacy

Nika Nusdorfer (Avtor), Aleš Obreza (Mentor), Marko Jukič (Komentor)

Povzetek

Vzdrževanje ravnotežja med sintezo in razgradnjo proteinov v telesu je nujno za njegovo pravilno delovanje. Proteasom je ključni mehanizem v telesu, ki skrbi za optimalno razgradnjo znotrajceličnih proteinov. Konstitutivni proteasom je izražen v večini celic, imunoproteasom pa le v celicah imunskega sistema. Strukturno sta si oba tipa proteasoma zelo podobna, razlikujeta se le v treh katalitičnih podenotah β. Trenutno so na trgu trije neselektivni zaviralci proteasoma, težava katerih so številni neželeni učinki. Načrtovanje in sinteza selektivnih zaviralcev imunoproteasoma bi lahko pomenila nove načine zdravljenja avtoimunskih in rakavih obolenj. Cilj magistrske naloge je bil načrtovanje in sinteza selektivnih kovalentnih zaviralcev imunoproteasoma, ki bi se vezali na aktivno mesto v katalitični podenoti LMP-7. Želeli smo, da se spojine najprej nekovalentno vežejo na podenoto LMP-7 in se tako približajo aktivnemu mestu treoninske proteaze, nato pa bi tvorili kovalentno vez s treoninom preko elektrofilne »bojne glave«. Odločili smo se za uvedbo kloroacetamidnih, metiloksiranskih in epoksiketonskih elektrofilnih centrov na substituiran aminopiperidinski skelet. V vlogi izhodnih spojin smo imeli Boc-(terc-butiloksikarbonil)-zaščitena 3-aminopiperidin in 4-aminopiperidin. Osnovni mehanizem večine reakcij je bila nukleofilna substitucija, izjema je reakcija acidolize. Preden smo odstranili Boc-zaščitno skupino, smo na prostem dušiku tvorili amid s pomočjo kislinskih kloridov in sklopitvenih reagentov. Aminopiperidinski skelet, zaščiten z Boc, smo substituirali z bifenil-4-karbonil kloridom, sintetizirali pa smo tudi aminopiperidinski skelet, na katerega smo pripeli 4-morfolinobenzojsko kislino z uporabo sklopitvenih reagentov. Nato smo spojinam odstranili Boc-zaščito in na prosto aminsko skupino pripeli tri različne elektrofilne centre, ki so se prek amidne vezi povezali na aminopiperidinski skelet. Uspešno smo sintetizirali štiri spojine s kloroacetamidno »bojno glavo«, eno z metiloksiransko »bojno glavo« in tri zaviralce z epoksiketonsko »bojno glavo«. Vseh osem končnih spojin bodo biokemijsko vrednotili na izolirani podenoti LMP-7 imunoproteasoma.

Ključne besede

imunoproteasom;aminopiperidin;selektivni zaviralci;podenota LMP-7;kovalentni zaviralci proteasoma;sinteza proteinov;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2018
Tipologija:	2.09 - Magistrsko delo
Organizacija:	UL FFA - Fakulteta za farmacijo
Založnik:	[N. Nusdorfer]
UDK:	543.057:547.96(043.3)
COBISS:	4658545
Št. ogledov:	430
Št. prenosov:	204
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Synthesis of covalent immunoproteasome inhibitors with amino-substituted piperidine ring
Sekundarni povzetek:	Maintaining normal cell homeostasis between synthesis and degradation of proteins is crucial for the proper functioning of the body. The proteasome is a key mechanism in the eukaryotic organisms that ensures optimum degradation of intracellular proteins. The constitutive proteasome is expressed in most eukaryotic cells whereas immunoproteasome is only expressed in the cells of the immune system. Structurally, both types of proteasomes are very similar, differing only in three catalytic β subunits. At the moment, there are three non-selective proteasome inhibitors approved by the FDA; the main problem of these inhibitors is the high presence of adverse effects. Designing and synthesis of selective immunoproteasome inhibitors would allow for new therapeutic possibilities in the treatment of autoimmune disorders and cancer therapy.The purpose of the master thesis was the design and synthesis of selective covalent immunoproteasome inhibitors, that would bind to the active site of threonine in a catalytic subunit LMP-7. We wanted the compounds first to bind non-covalently to the subunit LMP-7 and thus approach the active site of threonine protease. Then, the inhibitors would covalently bind the electrophilic "warhead" to the catalytic theronine. We decided to introduce chloroacetamide, methyleneoxirane and epoxyketone electrophilic centers to a substituted aminopiperidine skeleton. Our starting compounds were Boc protected 3-aminopiperidine and 4-aminopiperidine. The fundamental mechanism of most reactions was nucleophilic substitution, except for acidolysis. Before removing the Boc protecting group, an amide was formed on the free nitrogen with the use of acid chlorides and coupling reagents. The aminopiperidine skeleton protected with Boc was substituted with biphenyl-4-carbonyl chloride. We also synthesized the aminopiperidine skeleton to which 4-morpholinobenzoic acid was added using coupling reagents. Subsequently, Boc-protecting group was removed from the compounds and three free electrophilic centers joined to the free amine group through the amide bond. We successfully synthesized four compounds with a chloroacetamide "warhead", one with an ethylenoxirane "warhead" and three epoxyketones "warhead" inhibitors. All eight compounds will be biochemically evaluated on isolated subunits LMP-7 of immunoproteasome.
Sekundarne ključne besede:	immunoproteasome aminopiperidine selective inhibitors LMP-7 subunit;
Vrsta dela (COBISS):	Magistrsko delo/naloga
Komentar na gradivo:	Univ. Ljubljana, Fakulteta za farmacijo
Strani:	IX, 51 f.
ID:	12040859