Sinteza pirimidinskih zaviralcev interakcije med stresnim proteinom Hsp90 in košaperonom Cdc37

Gaberc, Tjaša (avtor), Tomašič, Tihomir (mentor), Dernovšek, Jaka (komentor)

Rak predstavlja enega izmed največjih globalnih zdravstvenih problemov. Zanj je med drugim značilna povišana koncentracija stresnih proteinov toplotnega šoka (Hsp), ki v celicah pomagajo pri pravilnem zvijanju proteinov do njihove nativne konformacije. Proteinska homeostaza je ključna za celično proliferacijo, zato je zaviranje delovanja Hsp zanimiv terapevtski pristop za zdravljenje raka. Med tovrstne Hsp-je sodi tudi protein Hsp90, katerega izražanje se pri raku močno poveča in njegovo zaviranje močno vpliva na sposobnost preživetja rakavih celic. Eden izmed možnih načinov zaviranja Hsp90 je zaviranje medproteinske interakcije med Hsp90 in njegovim košaperonom Cdc37, ki je v večini odgovoren za dostavljanje kinaz do Hsp90. Namen naloge je bil optimizacija predhodno sintetiziranih spojin raziskovalcev Katedre za farmacevtsko kemijo UL FFA, z željenim zaviranjem interakcije med proteinoma Hsp90 in Cdc37. Istočasno smo želeli vrednotiti odnos med strukturnimi lastnostmi naših ligandov in njihovim antiproliferativnim učinkom na rakavi celični liniji MCF-7. V ta namen smo sintetizirali devet novih, na pirimidinu osnovanih spojin. Načrtovali smo jih iz osnovnega N-metil-N-fenetilbenzamidnega skeleta, ki se je nadaljeval v pirimidinski skelet. Osnova za optimizacijo sta bili predhodno sintetizirani spojini TNG-116 in TNG-112, kjer smo namesto distančnika CH2-CH2-O med N-metilamidom in benzenom vezali distančnik CH2-CH2. Zasnovali smo dve knjižnici spojin, s katerimi smo vrednotili vpliv substitucije para in meta osrednjega benzenovega obroča na delovanje analogov in njihovo prileganje v vezavno mesto. Pri obeh knjižnicah smo istočasno na mestu 4 pirimidinskega obroča uvajali različne alifatske heterocikle in raziskovali vpliv njihove menjave. Z biološkimi testiranji na rakavi celični liniji MCF-7 smo odkrili pomembnost tvorbe dodatne akceptorske vodikove vezi na vmesniku CH2-CH2-O. Ugotovili smo, da funkcionalne skupine na mestu 4 pirimidinskega heterocikla z vezavnim mestom najverjetneje ne tvorijo ionske interakcije, medtem ko k vezavi na istem mestu pripomore možnost tvorbe dodatnih akceptorskih vodikovih vezi. Sklepamo tudi na neugodnost prekomernega povečanja velikosti alifatskega heterocikla in substitucije meta na benzenovem obroču. Izmed novo sintetiziranih spojin je N-metil-N-fenetil-3-((6-(pirolidin-1-il)pirimidin-4-il)amino)benzamid (9e) dosegla najnižjo srednjo zaviralno vrednost IC50 29 μM in tako predstavlja eno izmed izhodišč za nadaljnjo optimizacijo.

• Jezik: Slovenski jezik
• Ključne besede: Hsp90, rak, zaviralci medproteinske interakcije med Hsp90 in Cdc37, Cdc37, pirimidinski skelet
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2024
• PID: 20.500.12556/RUL-158946
• Datum objave v RUL: 23.06.2024

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Synthesis of pyrimidine-based heat shock protein 90 and co-chaperone Cdc37 interaction inhibitors

Izvleček

Cancer represents one of the greatest global health problems. Among other characteristics, it is marked by an elevated concentration of heat shock proteins (Hsp), which help in the correct folding of client proteins to their native conformation within cells. Protein homeostasis is crucial for cell growth and reproduction, making the inhibition of Hsp activity an intriguing therapeutic approach for cancer treatment. One such Hsp is the protein Hsp90, whose expression is significantly increased in cancer, and whose inhibition greatly impacts the survival ability of cancer cells. One of the possible ways to inhibit Hsp90 includes the inhibition of protein-protein interaction between Hsp90 and its cochaperone Cdc37, which is largely responsible for delivering kinases to Hsp90. The master's thesis aimed to optimize previously synthesized compounds by researchers at the Department of Pharmaceutical Chemistry, UL FFA, with the desired inhibition of the interaction between the proteins Hsp90 and Cdc37. At the same time, we wanted to evaluate the relationship between the structural properties of our ligands and their antiproliferative effect on the MCF-7 cancer cell line. For this purpose, we synthesized nine new pyrimidine-based compounds. We designed them from the basic N-methyl-N-phenethylbenzamide skeleton, which continued into the pyrimidine skeleton. The basis for the optimization were the previously synthesized compounds TNG-116 and TNG-112, where instead of a CH2-CH2-O linker between the N-methylamide and benzene, we inserted a CH2-CH2 linker. To evaluate the influence of para and meta substitution of the central benzene ring on the activity of the analogs and their fitting into the binding site, we designed two libraries of compounds. In both libraries, we simultaneously introduced various aliphatic heterocycles at position 4 of the pyrimidine ring and investigated the impact of their substitution. Biological testing on the MCF-7 cells revealed the importance of an additional hydrogen bond acceptor at the CH2-CH2-O linker. We found that the functional groups of the compound at position 4 of the pyrimidine heterocycle are unlikely to form ionic interactions with the binding site, whereas the ability to form additional acceptor hydrogen bonds contributes to binding at the same site. We also concluded that excessively increasing the size of the aliphatic heterocycle and meta substitution on the benzene ring are unfavorable. Among the newly synthesized compounds, N-methyl-N-phenethyl-3-((6-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)benzamide (9e) achieved the lowest mean inhibitory value IC50 of 29 μM, which thus serves as one of the starting points for further optimization.

• Ključne besede: Hsp90, cancer, inhibitors of protein-protein interaction between Hsp90 and Cdc37, Cdc37, pyrimidine scaffold