20.500.12556/RUL-114393 Sinteza zaviralcev encima InhA 1,2,3,4-tetrahidropirolo[1,2-a]pirazinskega tipa Synthesis of 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine type InhA inhibitors Tuberkuloza je nalezljiva bolezen, ki jo povzroča bakterija Mycobacterium tuberculosis, in ostaja vodilni vzrok smrti predvsem v nerazvitem delu sveta. Zdravilo prve linije kombiniranega zdravljenja predstavlja izoniazid, ki moti biosintezo mikolnih kislin, tako da zavira encim InhA. Zaviranje tega encima je povezano s celično smrtjo mikobakterije. Izoniazid je predzdravilo, ki potrebuje oksidativno aktivacijo z mikobakterijsko katalazno peroksidazo KatG. Monorezistentnost proti izoniazidu je najpogostejša odpornost proti zdravilu prvega izbora za terapijo tuberkuloze. Zato so učinkovine, ki neposredno zavirajo InhA in ne potrebujejo aktivacije s KatG, zelo obetavne za spopad z odpornimi sevi M. tuberculosis. Sintetizirati smo želeli štiri nove potencialne neposredne zaviralce encima InhA (spojine 8, 12, 17 in 26). Izhajali smo iz strukture molekule tetrahidropiranskega zaviralca (spojina I), ki smo mu tetrahidropiranski del zamenjali z 1,2,3,4-tetrahidropirolo[1,2-a]pirazinom. Kemijski prostor v aktivnem mestu encima ter njegovo zaviranje smo raziskali z različnimi substituenti na osnovni strukturi našega potencialnega zaviralca. Sintetiziranim spojinam so na Fakulteti za farmacijo Univerze v Ljubljani na izoliranem encimu InhA določili vrednosti IC50. Te so: za spojino 8 70 µM in za spojino 26 20 µM. Spojina 17 je razpadla, preden smo jo uspeli testirati. Tako smo tudi ugotovili, da je nesubstituiran pirol nestabilen, zato bi morali na obroč pripeti elektron-privlačne skupine. Slednji pristop smo preskusili pri spojini 12, kjer smo želeli na pirol uvesti brom, a žal želenega produkta nismo uspeli izolirati. Ne glede na to predvidevamo, da spojina 12 ne bi bila aktivna, pač pa bi imela le povečano kemijsko stabilnost. Tuberculosis is an infectious disease which is caused by Mycobacterium tuberculosis, and remains the leading cause of death in undeveloped countries. The first-line drug for antituberculotic treatment is isoniazid. It disrupts the biosynthesis of mycolic acids by inhibiting the enzyme InhA. Inhibition of this enzyme causes mycobacterium cell death. Isoniazid is a prodrug that needs oxidative activation with mycobacterium catalase-peroxidase KatG. Mono-resistance against isoniazid is the most common resistance against first-line antituberculotic agent. Thus, the direct inhibitors of InhA, that do not need activation by KatG, are very promising to combat resistant M. tuberculosis strains. We tried to synthesize four new potential direct inhibitors of InhA enzyme (compounds 8, 12, 17 in 26). Our synthesis was based on tetrahydropyran inhibitor molecule structure (compound I), in which we substituted the tetrahydropyran part with the more rigid 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine. With the introduction of different substituents on to 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine we researched the chemical space of InhA active site. The IC50 values of synthesized compounds were determined on the Faculty of Pharmacy, University of Ljubljana, on an isolated InhA enzyme. These values are 70 µM and 20 µM for compounds 8 and 26, respectively. Compound 17 degraded before we managed to test its activity. HRMS analysis of synthesized compound confirmed that unsubstituted pyrrole is unstable and is oxidized when in contact with oxygen. Electron-withdrawing substituents should be introduced to the pyrrole ring in order to increase chemical stability. The attempt to introduce bromine to the pyrrole ring was however not successful. We presume the product of the bromination would not be active, however it should be more chemically stable. tuberkuloza Mycobacterium tuberculosis neposredni zaviralci InhA 1 2 3 4-tetrahidropirolo[1 2-a]pirazin tuberculosis Mycobacterium tuberculosis direct InhA inhibitors 1 2 3 4-tetrahydropyrrolo[1 2-a]pyrazine true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2020-02-26 08:35:28 2020-02-26 08:35:35 2022-08-26 03:50:16 0000-00-00 00:00:00 2020 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 72783 916.pdf 916.pdf 1 4DB71A4529F0E891D770575F1DA3F13A b2322eb954864bf56f731f6900da7def62a663b98fb52963d87853a321b48503 35702dff-a1b8-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=126566 Fakulteta za farmacijo 0 0 0