Sinteza in vrednotenje novih N-alkil amidov kot potencialnih selektivnih zaviralcev butirilholin esteraze

Petrijan, Rok (author), Gobec, Stanislav (mentor), Grošelj, Uroš (comentor)

Raziskovanje novih zdravilnih učinkovin za zdravljenje Alzheimerjeve bolezni (AB) kot vodilnega vzroka demence pridobiva na pomenu. Obstoječa zdravila za simptomatsko zdravljenje AB, ki v večini spadajo v skupino zaviralcev encima acetilholin esteraze (AChE), delujejo kratkotrajno ter so primerna predvsem za zdravljenje v zgodnjih fazah bolezni. Pri napredovani obliki AB pride do povišanja aktivnosti in koncentracije encima butirilholin esteraze (BChE), ki je tako obetavna tarča za načrtovanje novih učinkovin za lajšanje simptomov v poznejših fazah AB. V magistrski nalogi smo se osredotočili na raziskovanje novih učinkovin, ki so selektivne za BChE in posledično izkazujejo možnost zdravljenja tudi v kasnejših fazah bolezni. V ta namen smo sintetizirali različne ciljne amide, ki so potencialni selektivni zaviralci encima BChE. Te smo sintetizirali na podlagi predlagane knjižnice amidov, ki je bila generirana na osnovi spojine vodnice z molekulskim sidranjem. Kot izhodiščno molekulo smo uporabili aminokislino L-triptofan, pri kateri smo na amino skupino uvedli različno dolge alkilne skupine, s karboksilno skupino pa smo tvorili amidno vez s pripenjanjem izbranih aminov. Pripravljene spojine smo v celoti okarakterizirali in ovrednotili njihovo aktivnost in vitro na encimih hBChE in hAChE z uporabo Ellmanove metode. Sintetizirali smo 11 novih za BChE selektivnih triptofanskih α-aminoamidov s srednjo zaviralno koncentracijo v nanomolarnem območju. Najboljšo zaviralno aktivnost so imele spojine z etilnim distančnikom med cikloalkilnim obročem in amidno skupino na karboksilnem delu ter n-butilno ročko na aminskem delu L-triptofana. V seriji novih sintetiziranih zaviralcev je bila najmočnejši zaviralec spojina 11 s 23,1 nM srednjo zaviralno koncentracijo. Zaradi preproste sinteze in dobljenih rezultatov biološkega testiranja lahko potrdimo, da spojine s triptofanskim ogrodjem predstavljajo primerno podlago za nadaljnjo optimizacijo do spojin vodnic pri načrtovanju novih zaviralcev BChE.

• Language: Slovenian
• Keywords: Alzheimerjeva bolezen, butirilholin esteraza, zaviralci butirilholin esteraze, triptofan
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2020
• PID: 20.500.12556/RUL-113441
• Publication date in RUL: 07.01.2020

Secondary language

• Language: English
• Title: Synthesis and evaluation of new N-alkyl amides as potential selective inhibitors of butyrylcholinesterase

Abstract

Research of new active substances for the treatment of Alzheimer's disease (AD), a leading cause of dementia, is gaining an importance. Existing symptomatic treatments, mostly consisting of acetylcholineesterase (AChE) inhibitors, have a short-term effect and are particularly suitable for treatment of early stages of the disease. With progression of AD, there is an increase in activity and concentration of the butyrylcholineesterase (BChE), which is therefore a promising target for the design of new active substances for symptomatic treatment in the later stages of AD. In this master's thesis we have focused on the research of novel agents that are selective for BChE and consequently demonstrate the possibility of treatment in the later stages of the disease. For that purpose, we synthesized various target amides, which are selective inhibitors of the BChE. These were synthesized on the basis of a proposed library of target amides, which was generated using lead compound and computational virtual screening. The amino acid L-tryptophan was used as the starting molecule, to which various long alkyl groups were attached to the amine moiety. By attachment of selected amines, an amide bond was formed using the carboxyl group. The prepared compounds were fully characterized and their in vitro activity was evaluated on hBChE and hAChE enzymes using the Ellman method. We synthesized 11 novel BChE-selective tryptophan α-aminoamides with a mean inhibitory concentrations in the nanomolar range. The compounds with the ethyl spacer between the simple cycloalkyl ring and the amide group on the carboxyl moiety, and the n-butyl handle on the amine moiety of L-tryptophan, showed the best inhibitory activity. In the series of new inhibitors, the strongest inhibitor was compound 11 with IC50 value of 23.1 nM. Due to the simple synthesis and promising results of the bioassay, we can confirm that the tryptophan-based compounds provide a suitable basis for further optimization as lead compounds in the design of new BChE inhibitors.

• Keywords: Alzheimer's disease, butyrylcholineesterase, butyrylcholineesterase inhibitors, tryptophan