Načrtovanje in sinteza dvojnih zaviralcev kinaze p38ɑ MAPK in butirilholin esteraze s 4-(naftalen-2-il)-3-(piridin-4-il)piridazinskim skeletom

Skenderović, Enita (author), Knez, Damijan (mentor), Meden, Anže (comentor)

Alzheimerjeva bolezen (AB) je kompleksna nevrodegenerativna bolezen, za katero je značilno postopno upadanje kognitivnih funkcij. Tipične spremembe v centralnem živčnem sistemu so nalaganje amiloidnih plakov in nevrofibrilarnih pentelj, ki pa ju med drugim spremljata propad holinergičnih, adrenergičnih in serotoninergičnih nevronov ter nevrovnetje. Pri našem delu smo se osredotočili na dve tarči: encim butirilholin esteraza (BChE) ter z mitogenom aktivirano proteinsko kinazo p38α (p38α MAPK). Zaviranje BChE izboljša kognitivne funkcije zaradi povečanja koncentracije živčnega prenašalca acetilholina, zaviranje kinaze p38α MAPK pa zmanjša nevrovnetje in hiperfosforilacijo proteina tau, kar bi lahko upočasnilo potek AB. Dvojne zaviralce BChE in p38α MAPK smo načrtovali na podlagi znanega selektivnega zaviralca kinaze p38α MAPK MW150 – (6-(4-metilpiperazin-1-il)-3-(naftalen-2-il)-4-(piridin-4-il)piridazina. Iz razrešene kristalne strukture spojine MW150 v kompleksu s p38α MAPK je razvidno, da je N-metilpiperazin usmerjen proti topilu in ga lahko enostavno nadomestimo s fragmenti, ki bi lahko v molekulo vnesli zaviralno delovanje na BChE. Za dosego dvojnega delovanja smo tako v molekulo uvedli N,N-dimetilaminoetilni fragment. Na podlagi literaturnih virov smo postavili sintezno pot, pri čemer smo izhajali iz γ-pikolina in naftalen-2-karboksilne kisline, ki smo ju v zaporednih sinteznih korakih pretvorili do 6-kloropiridazina 7, ki je služil kot sinton za sintezo dvojnih zaviralcev. Vzporedno smo postavili tudi sintezno pot za pripravo pirazolov, kjer je bil centralni piridazinski obroč nadomeščen s petčlenskim pirazolom. Dvojne zaviralce in kontrolne spojine smo ovrednotili v biokemijskem testu na izoliranih encimih. Izmed vseh sintetiziranih spojin je najbolj perspektiven zaviralec spojina 9, ki z IC50 vrednostjo 0,5296 ± 0,0595 µM zavira hBChE, v nizko nanomolarnem območju pa je zavirala tudi kinazo p38α MAPK (IC50 = 0,046 ± 0,011 µM). Spojina ima glede na rezultate računalniške analize fizikalno-kemijskih lastnosti in učinkovinam podobnih lastnosti tudi ustrezne karakteristike, ki napovedujejo dobro biološko uporabnost in prehajanje v centralni živčni sistem. Nadaljnje delo na seriji dvojnih zaviralcev bo usmerjeno v optimizacijo aktivnosti na obeh tarčnih encimih, saj je zaželeno, da sta aktivnosti na obeh tarčah uravnoteženi.

• Language: Slovenian
• Keywords: Alzheimerjeva bolezen, butirilholin esteraza, dvojni zaviralci, z mitogenom aktivirana proteinska kinaza p38α
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Publication date: 30.09.2023
• Year: 2022
• PID: 20.500.12556/RUL-141597
• Publication date in RUL: 01.10.2022

Secondary language

• Language: English
• Title: Design and synthesis of p38ɑ MAPK kinase and butyrylcholinesterase dual inhibitors with 4-(naphthalene-2-yl)-3-(pyridin-4-yl)piridazine scaffold

Abstract

Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by a gradual decline in cognitive function. Typical changes in the central nervous system include the accumulation of amyloid plaques and neurofibrillary tangles associated with the breakdown of cholinergic, adrenergic, and serotonergic neurotransmission and neuroinflammation, among others. In our work, we focused on two targets: the butyrylcholinesterase enzyme (BChE) and the mitogen activated protein kinase p38α (p38α MAPK). Inhibition of BChE improves cognitive function by increasing the concentration of the neurotransmitter acetylcholine, and inhibition of p38α MAPK kinase reduces neuroinflammation and hyperphosphorylation of tau protein, which may overall halt the progression of AD. Dual BChE and p38α MAPK inhibitors were developed based on the known selective p38α MAPK kinase inhibitor MW150 – (6-(4-methylpiperazin-1-yl)-3-(naphthalen-2-yl)-4-(pyridin-4-yl)pyridazine. As can be seen from the crystal structure of compound MW150 in complex with p38α MAPK, N-methylpiperazine is directed towards the solvent and can be easily replaced by fragments that could impart BChE inhibition. To achieve inhibition of both targets, N,N-dimethylaminoethyl substituent was introduced. Based on the literature, we developed a synthetic strategy starting from γ-picoline and naphthalene-2-carboxylic acid, which were converted in successive synthetic steps to 6-chloropyridazine 7, which served as a synthon for the synthesis of the dual inhibitors. In parallel, we also developed a synthetic route for the preparation of pyrazoles in which the central pyridazine ring was replaced by a five-membered pyrazole. The dual inhibitors and the control compounds were evaluated in a biochemical assay on isolated enzymes. Among all the synthesized compounds, compound 9 is the most promising inhibitor inhibiting hBChE and p38α MAPK kinase with IC50 values of 0.5296 ± 0.0595 µM and 0.046 ± 0.011 µM, respectively. According to the computational analysis of physicochemical properties and drug-like characteristics, compound 9 also exhibits suitable features suggesting good bioavailability and distribution in the central nervous system. Further work on the series of dual inhibitors aims to optimize the activity on both target enzymes to achieve balanced inhibitory potency.

• Keywords: Alzheimer's disease, butyrylcholinesterase, dual inhibitors, mitogen activated protein kinase p38α.