Sinteza cikličnih aminskih derivatov izoksazolo[5,4-d]pirimidina z delovanjem na Tollu podobne receptorje 7

Oman, Katja (avtor), Knez, Damijan (mentor), Sova, Matej (komentor)

Tollu podobni receptorji (TLR) igrajo pomembno vlogo v prirojenem in pridobljenem imunskem sistemu, ki je prva obrambna linija telesa pred mikrobi. TLR prepoznavajo molekulske vzorce, povezane s patogeni in poškodbami in sprožijo hiter imunski vnetni odziv. Trenutno je pri človeku znanih 10 podtipov TLR, mi smo se v sklopu našega dela osredotočili na TLR7, endosomski receptor, ki prepoznava enoverižno RNA virusov. Imunski sistem, aktiviran preko receptorja TLR7, privede do protivnetnega, protivirusnega in protitumornega učinka, zato bi lahko bili agonisti receptorja TLR7 uporabni pri zdravljenju rakavih obolenj, astme in virusnih okužb. Tekom magistrske naloge smo sintetizirali analoge agonista TLR7. Pripravljene spojine smo okarakterizirali v bioloških testiranjih, s čimer smo pridobili nekaj novih informacij o odnosu med strukturo in delovanjem agonistov TLR7 na osnovi izoksazolo[5,4-d]pirimidina. V sklopu šeststopenjske sinteze smo iz 4-fluorbenzaldehida preko oksima, imidoil klorida, derivata izoksazola ter nato izoksazolo[5,4-d]pirimidin-4(5H)-ona in njegove pretvorbe v aril klorid, na mesto 4 heterobicikličnega ogrodja pripeli različne substituente in sintetizirali šest končnih spojin, potencialnih agonistov TLR7. Za pet spojin smo nato ocenili topnost v celičnem mediju, preverili njihovo agonistično delovanje na TLR7, za nekatere tudi na TLR8, ter za najboljšo še citotoksičnost na celični liniji HEK293. Vse spojine so bile zaradi lipofilnih fragmentov slabo topne v celičnem mediju. Spojina 12 je edina izkazovala agonistično delovanje na TLR7 z EC50 vrednostjo 15,9 ± 0,9 μM. V primerjavi z imikvimodom, trenutno edinim registriranim agonistom TLR7 (EC50 = 2,1 µM), je vrednost EC50 spojine 12 višja, kar kaže na njeno šibkejše agonistično delovanje. Zaključimo lahko, da spojina 12 predstavlja dobro izhodišče za nadaljnje raziskave novih agonistov TLR7 kot imunomodulatornih učinkovin.

• Jezik: Slovenski jezik
• Ključne besede: Tollu podobni receptor, TLR7, imunski sistem, rak, sinteza
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2023
• PID: 20.500.12556/RUL-152654
• Datum objave v RUL: 02.12.2023

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Synthesis of cyclic amine derivatives of isoxazolo[5,4-d]pyrimidine to target Toll-like receptors 7

Izvleček

Toll-like receptors (TLRs) play an important role in the innate and adaptive immune system, which is the first line of defense against microbes. TLR7 receptors recognize molecular patterns associated with pathogens and damage and trigger a rapid inflammatory immune response. Currently, 10 TLR receptor subtypes are known in humans, and in this work we have focused on TLR7, an endosomal receptor that recognizes viral single-stranded RNA. The immune system activated by TLR7 leads to anti-inflammatory, antiviral and antitumor activity, which is why TLR7 agonists could be useful in the treatment of cancer, asthma and viral infections. As part of this master's thesis, we synthesized analogs of TLR7 agonists. The compounds were evaluated in biological assays to obtain additional data on the structure-activity relationships of isoxazolo[5,4-d]pyrimidine-based TLR7 agonists. The six-step synthesis, starting from 4-fluorobenzaldehyde, its conversion to into oxime, imidoyl chloride, isoxazole, isoxazolo[5,4-d]pyrimidin-4(5H)-one and then into aryl chloride, allowed us to attach different substituents at position 4 of the heterobicyclic core and to synthesize six final compounds, potential TLR7 agonists. The solubility of five derivatives in cell media was estimated, followed by the evaluation of TLR7 agonism, TLR8 agonism for certain analogs, and cytotoxicity on the HEK293 cell line for the most potent compound. All compounds were poorly soluble in the cell medium due to the lipophilic fragments. Compound 12 was the only compound that showed agonism on TLR7 with an EC50 value of 15.9 ± 0.9 µM. Compared to imiquimod, the only approved TLR7 agonist (EC50 = 2.1 µM), the EC50 of compound 12 is higher, indicating its weaker agonist activity. In summary, compound 12 represents a good starting point for further research into novel TLR7 agonists as immunomodulatory agents.

• Ključne besede: Toll-like receptors, TLR7, immune system, cancer, synthesis