20.500.12556/RUL-166367 Sinteza in vrednotenje indolin-1-karbonil fluoridov kot kovalentnih zaviralcev kaspaze 1 Synthesis and evaluation of indoline-1-carbonyl fluorides as covalent caspase 1 inhibitors Alzheimerjeva bolezen (AB) je nevrodegenerativna bolezen, ki vodi v postopno izgubo spomina in kognitivnih funkcij – demenco. Njene ključne patološke značilnosti vključujejo kopičenje amiloidnih plakov in nevrofibrilarnih pentelj v možganih. V zadnjem času se vedno bolj prepoznava vloga nevrovnetja kot pomembnega dejavnika pri nastanku in napredovanju AB. Imunske celice centralnega živčnega sistema ob zaznavi patoloških signalov aktivirajo inflamasom, ki nadalje sproži imunski odziv: aktivira se kaspaza-1, cisteinska proteaza, ki aktivira provnetna citokina interlevkin-1β (IL-1β) in interlevkin-18 (IL-18), ki spodbudita vnetni odziv. Poleg tega kaspaza-1 cepi gasdermin D in sproži piroptozo, vnetno obliko celične smrti, ki povzroči propad živčnih celic. V magistrski nalogi smo načrtovali in sintetizirali kovalentne zaviralce kaspaze-1, ki bi preko zaviranja kaspaze-1 zavrli vnetje in piroptozo ter pripomogli k zaustavitvi nevrodegeneracije. Namen našega dela je torej sintetizirati nove zaviralce kaspaze-1, ki bodo lahko zapolnili to vrzel v obetavnem področju načrtovanja učinkovin, ki vplivajo na proces nevrovnetja. Uporabili smo pristop načrtovanja novih učinkovin na podlagi fragmentov in tehniko rasti fragmentov. Sintetizirali smo 16 kovalentnih zaviralcev kaspaze-1 in jih ovrednotili z biokemijskim testiranjem in vitro na človeški rekombinantni kaspazi-1. Osnovnemu, predhodno odkritemu zaviralcu kaspaze-1, indolin-1-karbonil fluoridu, smo na različna mesta na obroču pripenjali dodatne substituente po principu rasti fragmenta. Ugotovili smo, da substituenti na mestih 5 in 6 indolinskega obroča ključno vplivajo na zaviralno aktivnost, pri čemer so bolj ugodni manjši substituenti, kot sta nitro (27) in ciano skupina (29), prvi derivat je bil tudi najmočnejši zaviralec kaspaze-1. Spojine z večjimi substituenti na mestu 3, kot so terc-butil propanoat (spojina 25) ter terc-butil butanoat (26), so močni zaviralci kaspaze-1, karboksilata 43 in 44 pa sta šibkejša zaviralca. Za zaviralno aktivnost so neugodni tudi substituenti na mestu 7. Povzamemo lahko, da so za zaviralno aktivnost najbolj ugodni majhni substituenti za mestu 5, večji lipofilni fragmenti na mestu 3, pa tudi substituenti na mestu 6 (metil karboksilat). Z raziskovanjem kemijskega prostora indolin-1-karbonil fluoridov smo tako pridobili dodatne informacije, ki bodo uporabne pri nadaljnjem razvoju kovalentnih zaviralcev kaspaze-1 tega strukturnega podtipa. Alzheimer's disease (AD) is a neurodegenerative disease that leads to gradual loss of memory and cognitive functions – dementia. Its main pathological characteristics include the buildup of amyloid plaques and neurofibrillary tangles in the brain. Lately, the role of neuroinflammation has been increasingly recognized as an important factor in AD onset and progression. In response to pathological signals, immune cells of the central nervous system activate the inflammasome, which initiates the immune response: it activates caspase-1, a cysteine protease that activates pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18), which promote the inflammatory response. Additionally, caspase-1 cleaves gasdermin D, leading to pyroptosis, an inflammatory form of programmed cell death that results in neuronal loss. In this master's thesis, we designed and synthesized covalent inhibitors of caspase-1 intended to suppress inflammation and pyroptosis, thereby contributing to reducing neurodegeneration. The aim of our work is therefore to synthesize new caspase-1 inhibitors that will be able to fill this gap in the promising field of designing drugs that affect the neuroinflammation process. We used the principle of fragment-based drug discovery and the fragment growing technique to synthesize and evaluate 16 caspase-1 inhibitors using in vitro biochemical assay on human, recombinant caspase-1. To a previously identified caspase-1 inhibitor, indoline-1-carbonyl fluoride, additional substituents were attached at various positions on the indoline ring. We observed that substituents at positions 5 and 6 of the indoline ring have a crucial impact on the inhibitory potency, with smaller substituents, such as nitro (27) and cyano (29) groups, being particularly favourable. Compound 27 was identified as the most potent caspase-1 inhibitor. Compounds with larger substituents at position 3, such as tert-butyl propanoate (25) and tert-butyl butanoate (26), are potent caspase-1 inhibitors, while their carboxylate derivatives 43 and 44 inhibit caspase-1 to a lesser extent. Substituents at position 7 are also unfavourable, leading to weaker inhibitors. We can conclude that the most favourable modifications for inhibitory activity include small substituents at position 5, larger lipophilic fragments at position 3, and also substituents at position 6 (methyl carboxylate) of indoline core. By exploring the chemical space of indoline-1-carbonyl fluoride, we obtained additional information that will be valuable for the further development of caspase-1 inhibitors of this chemical subtype. kaspaza-1 karbamoil fluorid kovalentni zaviralci nevrovnetje Alzheimerjeva bolezen caspase-1 carbamoyl fluoride covalent inhibitors neuroinflammation Alzheimer's disease true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-01-09 07:45:05 2025-01-09 07:45:17 2025-01-10 03:53:34 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 114515 26806.pdf 26806.pdf 1 2464B7A05B86015F6BADB29E93F39C4C 6e581bff6af3a369a57fe3d19a1210657ab18cf134900f26b1a4be80f172a724 902402ad-ce54-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=197940 Fakulteta za farmacijo 0 0 0