Načrtovanje in sinteza novih antagonistov receptorja NOD2 na osnovi N-alkilbenzamidnega osrednjega ogrodja : enoviti magistrski študij farmacija

Slivnik, Matevž (author), Jakopin, Žiga (mentor)

Imunski sistem ščiti naše telo pred različnimi patogeni. Na grobo ga delimo na prirojeni in pridobljeni imunski sistem. Kljub temu da prirojeni imunski sistem ni zmožen antigen-specifičnega razlikovanja med patogeni, pa lahko prepozna tuje molekule s pomočjo receptorjev za prepoznavo vzorcev (PRR), ki prepoznajo molekulske vzorce značilne za patogene (PAMP) in molekulske vzorce značilne za nevarnost (DAMP). Ena izmed družin PRR-jev so NOD-u podobni receptorji (angl. Nod-like receptor — NLR), v katero sodita tudi receptorja NOD1 in NOD2. Gre za citosolna receptorja, ki prepoznavata citoplazemske PAMP. V primeru receptorja NOD1 je to iE-DAP, medtem ko je najmanjši fragment, ki ga prepozna receptor NOD2 MDP. Po prepoznavi liganda receptorja aktivirata signalne poti NF-κB, IRF in MAPK, kar sproži vnetni odziv. Receptor NOD2 kodira gen CARD15, različni polimorfizmi in mutacije na tem genu pa lahko spremenijo funkcijo NOD2, kar privede do kroničnih vnetnih, avtoimunskih in rakavih obolenj. Antagonisti NOD2 imajo zato terapevtski potencial pri zdravljenju bolezni, ki so posledica povečane signalizacije NOD2. V sklopu magistrskega dela smo sintetizirali 10 novih antagonistov NOD2. Kot spojino vodnico smo uporabili N-(2-(1-(2,3-dihidro-1H-inden-5-il)amino)-2-oksoetil)-1H-benzo[d]imidazol-2-il)metil)benzamid, pri kateri smo na mesto benzoilnega dela spojine uvedli različne substituente s katerimi smo vplivali na vezavo na receptor in/ali fizikalno-kemijske lastnosti. Za vse novo sintetizirane antagoniste so na Katedri za klinično biokemijo Fakultete za farmacijo Univerze v Ljubljani izvedli teste citotoksičnosti in ovrednotili antagonistično delovanje na receptorja NOD1 in NOD2. Rezultati testov so pokazali, da 3 spojine (spojine 5, 9 in 20) izkazujejo boljšo NOD2 antagonistično aktivnost v primerjavi s spojino vodnico, kot najboljši antagonist NOD2 naše serije spojin pa se je izkazala spojina 20. Večina novo sintetiziranih antagonistov NOD2 je imela primerljivo antagonistično delovanje tudi na receptor NOD1. Na osnovi rezultatov smo uspeli pridobiti nekaj informacij o odnosu med strukturo in delovanjem naših končnih spojin. Zamenjava benzoilnega dela spojine vodnice z arilsulfonilnim (spojini 5 in 9) ali benzilnim delom (spojina 20) je izboljšala NOD2 antagonistično aktivnost, medtem ko je prisotnost bazičnega centra pri spojinah 7, 11, 14, 16, 18 in 21 neugodno vplivala na NOD2 antagonistično aktivnost.

• Language: Slovenian
• Keywords: receptor NOD2 antagonisti NOD2 N-alkilbenzamid
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Place of publishing: Ljubljana
• Publisher: [M. Slivnik]
• Year: 2018
• Number of pages: IX, 71 f.
• PID: 20.500.12556/RUL-120278
• UDC: 543.057:612.017(043.3)
• COBISS.SI-ID: 4650865
• Publication date in RUL: 17.09.2020

Secondary language

• Language: English
• Title: Design and synthesis of novel NOD2 receptor antagonists based on N-alkylbenzamide central scaffold

Abstract

Immune system protects our body from various pathogens. We can roughly divide it in two groups: innate and adaptive immune system. Although the innate immune system is not capable of antigen-specific differentiation of pathogens, it can still recognize foreign molecules using the pattern recognition receptors (PRR), which recognize pathogen associated molecular patterns (PAMPs) and danger associated molecular patterns (DAMPs). NOD1 and NOD2 receptors belong to NLRs, which are one of the PRR families. NOD1 and NOD2 receptors are cytosolic receptors that recognize cytoplasmic PAMPs. In the case of NOD1 receptor that PAMP is iE-DAP, whereas MDP is the smallest fragment recognized by NOD2 receptor. After ligand recognition, the receptors activate NF-κB, IRF and MAPK signaling pathways, which triggers inflammatory response. NOD2 receptor is encoded by CARD15 gene. Different polymorphisms and mutations on that particular gene can change the NOD2 function, which can lead to chronic inflammatory and autoimmune diseases or cancer. Therefore, NOD2 antagonists have a therapeutic potential in treating diseases that occur as a result of an increased NOD2 signaling. In this master’s thesis, we synthesized 10 novel NOD2 antagonists. We used N-(2-(1-(2,3-dihydro-1H-inden-5-yl)amino)-2-oxoethyl)-1H-benzo[d]imidazol-2-yl)methyl)benzamide as our lead compound. We replaced the benzoyl moiety of the lead compound with various substituents, affecting the receptor binding and/or physicochemical properties. The Chair of Clinical Biochemistry, University Ljubljana, Faculty of pharmacy performed the cytotoxicity assay, NOD1 antagonistic activity evaluation and NOD2 antagonistic activity evaluation on newly synthesized antagonists. The results showed that three compounds (compounds 5, 9 and 20) expressed increased antagonistic activity compared to that of the lead compound with compound 20 being the best NOD2 antagonist of our compound series. Most of the newly synthesized NOD2 antagonists possessed similar NOD1 antagonistic activity. Based on the results, we were able to gather some information regarding the structure-activity relationship of our final compounds. The replacement of our lead compound’s benzoyl moiety with arylsulfonyl (compounds 5 in 9) or benzyl moiety (compound 20) improved the NOD2 antagonistic activity, while the presence of basic center in compounds 7, 11, 14, 16, 18 and 21 showed negative effect on the NOD2 antagonistic activity.