20.500.12556/RUL-121094 Načrtovanje, sinteza in vrednotenje derivatov gvanidina kot antagonistov Toll-u podobnega receptorja 4 [enoviti magistrski študij farmacija] Design, synthesis and evaluation of guanidine derivatives as antagonists of Toll like receptor 4 Toll-u podoben receptor 4 (TLR4) že vrsto let povezujejo s pojavom sepse, povzročene z vdorom Gram negativnih bakterij v človeško kri. Bolezen se v veliki meri pojavlja pri hospitaliziranih bolnikih, smrtnost zanjo pa je izredno visoka. Pri tem se poraja dosti vprašanj o načinu ter ustreznosti zdravljenja. Vse to je razlog za veliko povpraševanje po zdravilih za preprečevanje ali zdravljenje sepse, a kljub številnim raziskavam na tem področju, na tržišču še ni registriranega specifičnega zdravila. Antagonisti TLR4 predstavljajo velik potencial pri iskanju novih zdravil pri zdravljenju sepse. Toll-u podobni receptorji 4 igrajo pomembno vlogo v prirojenem imunskem sistemu, hkrati pa v manjši meri vplivajo tudi na pridobljeni imunski odziv posameznika. Gre za najbolj poznan in opisan transmembranski receptor iz skupine TLR. Značilno prepoznava molekulske vzorce povezane s patogeni, najpomembnejše pa je njegovo prepoznavanje lipopolisaharida, ki je glavna sestavina celične membrane Gram negativnih bakterij. V okviru magistrske naloge smo načrtovali sintezno pot za derivate spojine z dokazanim antagonističnim delovanjem na TLR4, ki je bila odkrita na Fakulteti za farmacijo v Ljubljani s pomočjo virtualnega rešetanja knjižnice spojin. Preizkusili smo več različnih sinteznih poti za pripravo gvanidinskih derivatov. Za najuspešnejšo pa se je izkazala tristopenjska sinteza, ki smo jo uporabili pri sintezi štirih končnih spojin. Sintetiziranim spojinam smo določili topnost v celičnem mediju, citotoksičnost na celični liniji HEK 293 ter ovrednotili antagonistično delovanje na celicah s selektivno izraženim TLR4. Izmed testiranih končnih spojin je ena izkazovala precej obetajoče antagonistično delovanje z določeno vrednostjo srednje inhibitorne koncentracije 27 μM. Za dodatne informacije o odnosu med strukturo in delovanjem spojin pa bi bilo potrebno sintetizirati še nekaj več derivatov. For many years Toll-like receptor 4 has been associated with the occurrence of sepsis, caused by intrusion of Gram negative bacteria into the human blood. The disease largely occurs in hospitalized patients and its mortality rate is extremely high, therefore it raises a lot of questions about methods and adequacy of the treatment. This is the reason for the high demand for medicinal products with the purpose of treatment or prevention of sepsis. However, despite numerous research in this area, a specific product has not been registered on the market. TLR4 antagonists present high potencial for the development of novel agents against sepsis. These receptors play an important role in the innate immune system and to a lesser extent also affect the adaptive immune response of an individual. TLR4 is well-known and described transmembrane receptor in the TLR group. It typically recognizes molecular patterns associated with pathogens, however, the most important is the recognition of lipopolysaccharide, which is a major component of the cell membrane of Gram negative bacteria. The aim of our master's thesis was to design the synthetic pathway for derivatives of a hit compound with proven antagonism of TLR4, which was discovered at the Faculty of Pharmacy in Ljubljana by virtual screening of the compound library. We examined several synthetic routes and finally discovered the three-stage synthesis of guanidine derivatives. Four final compounds were synthesized and evaluated. We have determined the solubility of compounds in the cell medium, the cytotoxicity in HEK 293 cell line and evaluated the antagonist activity of final compounds on cells which selectively express TLR4. Half maximal inhibitory concentration (IC50) of 27 μM has been determined for one of the final compounds, which shows promising antagonistic activity. However, it would be necessary to synthesize more derivates to obtain additional informations about structure-activity relationship. Toll-u podobni receptorji Toll-u podoben receptor 4 antagonisti TLR4 derivati gvanidina imunski odziv Toll-like receptors Toll-like receptor 4 TLR4 antagonists guanidine derivatives sepsis true false false [K. Rožman] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2020-09-30 08:17:46 2020-09-30 08:17:47 2022-09-01 04:01:31 0000-00-00 00:00:00 2014 0 Ljubljana 2014 Ljubljana VI, 52 f. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 535.33/.34+543.544+616.94(043.3) 3617393 MAG_Rozman_Kaja_2014.pdf MAG_Rozman_Kaja_2014.pdf 1 5F98FC015819A01448B93D09FDC8480D dc68d4bae8f1a019d94616343bdbd3d257e2034491f3f32b4fd65400c5c40fb7 394f4dd5-a1ba-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=135964 Fakulteta za farmacijo 0 0 0