Razvoj zaviralcev imunoproteasoma in modulatorjev imunskih signalnih poti s protitumornim in protivnetnim delovanjem

Strašek Benedik, Nika (avtor), Sosič, Izidor (mentor), Sova, Matej (komentor)

Imunski sistem ima ključno, a kompleksno vlogo pri zaščiti organizma. V določenih okoliščinah lahko postane prekomerno aktiven ali premalo odziven, kar vodi v razvoj kroničnih vnetij, avtoimunskih bolezni in rakavih obolenj. Z zaviranjem pretiranega imunskega odziva lahko zdravimo avtoimunske bolezni, medtem ko lahko z imunomodulatorji, ki dodatno aktivirajo imunski sistem, okrepimo odziv proti okužbam ali rakavim celicam. V našem delu smo se osredotočili na razvoj novih imunomodulatornih učinkovin, s katerimi smo ciljali različne farmakološke tarče vpletene v patogenezo omenjenih obolenj: TLR7 in 8, TAK1 ter imunoproteasom. Načrtovanja in optimizacije novih učinkovin smo se lotili z različnimi pristopi. Sintetizirali smo serijo spojin z izoksazolo[5,4-d]pirimidinskim skeletom, kjer so najbolje delujoči analogi izkazali selektivno agonistično delovanje na TLR7 v nizko mikromolarnih koncentracijah ter povečali sproščanje vnetnih citokinov. Z načrtovanjem na osnovi fragmentov smo pripravili obsežno knjižnico 2-cianobenzotiazolov, benzimidazolov ter 1-metilbenzimidazolov z različnimi elektron privlačnimi ter elektron donorskimi skupinami vezanimi na obroč. Določili smo reaktivnost in stabilnost sintetiziranih derivatov ter razvili analoge, ki bodo koristni za prihodnje aplikacije v biokonjugaciji in kemijski biologiji. Pripravljeni analogi so izkazali selektivno zaviralno aktivnost do podenote ß5i imunoproteasoma z vrednostmi IC50 v nizkem mikromolarnem območju. Prav tako so izkazali sposobnost znižanja sproščanja citokinov ter selektivno citotoksično delovanje na celice multiplega mieloma. Kot prvi smo načrtovali, sintetizirali ter biološko ovrednotili himerne razgrajevalce (PROTAC) TAK1, ki so povzročili razgradnjo TAK1 v s TNF-? stimulirani celični liniji raka dojke ter monocitni levkemični celični liniji. Spojini, ki sta izzvali najvišjo razgradnjo TAK1, sta izkazovali signifikanten vpliv na zmanjšanje živosti omenjenih celičnih linij stimuliranih s TNF-?, ter izzvali celično apoptozo. Selektivno delovanje molekul PROTAC le pod vnetnimi pogoji predstavlja velik potencial za nadaljnji razvoj selektivnih terapij za različna obolenja povezana z neustreznim imunskim odzivom. Na vseh treh področjih smo dosegli zadane cilje in razvili nove selektivne modulatorje z aktivnostmi v nizko mikromolarnih ter nanomolarnih koncentracijah.

• Jezik: Slovenski jezik
• Ključne besede: imunomodulatorji, Toll-u podobni receptorji, TAK1, imunoproteasom
• Vrsta gradiva: Doktorsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2024
• PID: 20.500.12556/RUL-165267
• Datum objave v RUL: 29.11.2024

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Development of immunoproteasome inhibitors and modulators of immune signaling pathways with anti-tumor and anti-inflammatory activity

Izvleček

The immune system plays an important but complex role in protecting the organism against patogens. Under certain circumstances, it can become overactive or underactive, which can lead to the development of chronic inflammation, autoimmune diseases, allergies, and cancer. Autoimmune diseases can be treated by suppressing an excessive immune response, while immunomodulators, which additionally activate the immune system, can strengthen the response against infections or cancer cells. In our work, we have focused on the development of new immunomodulatory agents for different pharmacological targets involved in the development of autoimmune diseases and cancers: TLR7 and 8, TAK1 and the immunoproteasome. We synthesized a series of compounds with an isoxazolo[5,4-d]pyrimidine scaffold, with the most potent analogs showing selective agonist activity on TLR7 at low micromolar concentrations and increasing the secretion of inflammatory cytokines. Using fragment-based design, we prepared a large library of 2-cyanobenzothiazoles, benzimidazoles and 1-methylbenzimidazoles with different electron-withdrawing and electron-donating substituents. We determined the reactivity and stability of synthesized derivatives and developed analogs that will be useful for future applications in bioconjugation and chemical biology. The prepared analogs showed selective inhibitory activity against the β5i-subunit of the immunoproteasome with IC50 values in the low micromolar range. They also showed a reduction in the release of cytokines and selective cytotoxicity on multiple myeloma cells. We were the first to develop, synthesize, and biologically study chimeric TAK1 degraders (TAK1 PROTACs), which induced TAK1 degradation in a TNF-α-stimulated breast cancer cell line and a monocytic leukemia cell line. The compounds that induced the strongest degradation of TAK1 showed a significant effect on reducing the viability of the aforementioned TNF-α-stimulated cell lines and caused cell apoptosis. The selective effect of PROTACs only under inflammatory conditions represents great potential for the further development of selective therapies for various conditions related to the improper immune response. To conclude, we achieved our goals in all three areas, and developed new selective modulators with activities at low micromolar and nanomolar concentrations.

• Ključne besede: immunomodulators, Toll-like receptors, TAK1, immunoproteasome