Sinteza 4-((3,5-dimetil-1H-pirazol-1-il)metil)benzamidnih zaviralcev encima InhA

Zajec, Živa (author), Hrast, Martina (mentor), Pajk, Stane (co-mentor)

Tuberkuloza je infekcijska bolezen, ki jo povzroča mikobakterija M. tuberculosis in najpo-gosteje prizadene pljuča. Tuberkuloza v svetovnem merilu predstavlja velik zdravstveni problem, saj zaradi nje vsako leto umre 1,3 milijona ljudi. Mikobakterijska celična stena je bogata z lipidi, ključni gradnik pa so mikolne kisline. Encimi, ki sodelujejo pri sintezi mi-kolnih kislin, so ravno zaradi tega glavne tarče učinkovin, ki jih uporabljamo za zdravljenje okužb z M.Tuberculosis. Encim InhA (enoil acilni prenašalni protein) sodeluje pri sintezi mikolnih kislin in je tarča izoniazida, ki je zdravilo prvega izbora. Izoniazid se mora za svoje delovanje aktivirati s peroksidazo KatG. Če pride na genu za KatG do mutacije, postane izoniazid neučinkovit. Zaradi vse pogostejšega pojava sevov, odpornih na izoniazid, razvi-jajo nove direktne zaviralce encima InhA, ki za svoje delovanje ne potrebujejo aktivacije. Eden izmed strukturnih razredov direktnih zaviralcev InhA, ki so jih odkrili z visoko zmo-gljivostnim rešetanjem in so izkazali dober potencial za nadaljnji razvoj, so benzamidni za-viralci encima InhA. V magistrski nalogi smo sintetizirali sedem benzamidnih zaviralcev InhA. V prvem delu magistrske naloge smo sintetizirali 4-((3,5-dimetil-1H-pirazol-1-il)metil)-N-(2-(4-feniltiazol-2-il)ciklopentil)benzamid in 4-((3,5-dimetil-1H-pirazol-1-il)metil)-N-(2-(4-feniltiazol-2-il)cikloheksil)benzamid. Obe molekuli imata kiralni center, zato smo sintezo načrtovali tako, da smo dobili dva diastereomera. Pri biološkem testiranju zaviralne aktivnosti se je izkazalo, da imajo cis in trans derivata spojine s petčlenskim obro-čem in spojina s šestčlenskim obročem zelo podobno zaviralno aktivnost v nizkem mikro-molarnem območju. V drugem delu magistrske naloge smo sintetizirali pet analogov 4-((3,5-dimetil-1H-pirazol-1-il)metil benzamidnih zaviralcev, tako da smo sistematično menja-li substituente, in sicer smo uvedli 2-fenilpiperidin, 2-fenilpirolidin, 2-fenilazetidin, 1-metil-3-fenilpiperazin in 3-fenilmorfolin. Trije analogi so izkazali zaviralno aktivnost v mikromo-larnem območju. Spojini trans 4-((3,5-dimetil-1H-pirazol-1-il)metil)-N-(2-(4-feniltiazol-2-il)ciklopentil)benzamid in (4-((3,5-dimetil-1H-pirazol-1-il)metil)fenil)(2-fenilpiperidin-1-il)metanon pa imata tudi zmerno protibakterijsko aktivnost. Z našim delom smo raziskali odnos med strukturo in delovanjem ter preverili, kako različni substituenti vplivajo na zavi-ranje encima InhA.

• Language: Slovenian
• Keywords: tuberkuloza, zaviralci encima InhA, benzamidni zviralci InhA, 4-((3, 5-dimetil-1H-pirazol-1-il)metilbenzamidi
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2019
• PID: 20.500.12556/RUL-108328
• Publication date in RUL: 28.06.2019

Secondary language

• Language: English
• Title: Synthesis of 4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide InhA inhibitors

Abstract

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis, which most commonly affects the lungs. Tuberculosis is a major healthcare burden causing 1.3 million deaths every year. Mycobacterial cell wall is rich in lipids, and the major part of lipid com-ponents are fatty acids called mycolic acids. Enzymes within mycolic acid synthesis path-way are drug targets. One of those enzymes is InhA (enoyl acyl carrier protein), an enzyme which is targeted by isoniazid, a drug of first choice for tuberculosis treatment. Isoniazid is a prodrug and requires activation by KatG peroxidase. Since KatG is essential for activa-tion, mutation on KatG gene causes resistance of M. tuberculosis to isoniazid. Therefore research is conducted on direct InhA inhibitors, which do not require activation. One of direct InhA inhibitor classes discovered through high through-put screening are benzamide inhibitors, which show a good potential for further optimization. In the master thesis our goal we synthesized seven benzamide inhibitors. Firstly, we syn-thesized compounds 4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)-N-((2-(4-phenylthiazol-2-yl)cyclopentyl)benzamide and 4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)-N-((2-(4-phenylthiazol-2-yl)cyclohexyl)benzamide. Since both compounds have a chiral center, we planed the synthesis to prepare both diastereomeres. Both diastereomeres of compound with cyclopentane ring and compound with cyclohexane ring exhibited very similar inhibi-tor potency in micro molar range. In the second part of the master thesis we focused on a synthesis of five different analogues of 4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl ben-zamide inhibitors. We used 2-phenilpiperidine, 2-phenilpirolidine, 2-phenilazetidine, 1-methyl-3-phenilpiperazine in 3-phenilmorpholine as substituents. Three of synthesized ana-logues exhibited inhibitor potency in low micro molar range. Compounds trans 4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)-N-((2-(4-phenylthiazol-2-yl)cyclopentyl)benzamide and (4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)phenyl)(2-phenylpiperidin-1-yl)methanone also showed modest antibacterial activity. In this thesis, we researched structure-activity rela-tionship between different analogues of benzamide InhA inhibitors and their potency.

• Keywords: tuberculosis, InhA inhibitors, benzamide InhA inhibitors, 4-((3, 5-dimethyl-1H-pyrazol-1-yl)methylbenzamides