o:38601 Dizajniranje, sinteza i ispitivanje bioloških osobina dualnih inhibitora ciklooksigenaze-2 (COX-2) i 5-lipoksigenaze (5-LOX) doktorska disertacija Design, synthesis and testing of biological properties of dual inhibitors of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) : doctoral dissertation sr Dualna inhibicija COX-2 i 5-LOX enzima predstavlja racionalnu strategiju razvoja potentnijih i bezbednijih antiinflamatornih lekova. Zapažena je prekomerna ekspresija COX-2 i 5-LOX enzima u tumorskim ćelijama, te se smatra da bi dualni inhibitori imali primenu u terapiji kancera. Prva faza istraživanja je dizajniranje novih, potencijalnih dualnih COX-2 i 5-LOX inhibitora primenom SAR analize i kompjuterskih metoda (3D-QSAR studija i molekulski doking). Sintetisano je i strukturno okarakterisano devetnaest jedinjenja, čija je aktivnost ispitana upotrebom komercijalno dostupnih COX-1, COX-2 i 5-LOX inhibitornih enzimskih eseja. Jedinjenja 1, 2, 3, 5, 6, 11 i 12 su se izdvojila kao dualni inhibitori COX-2 i 5-LOX enzima (COX-2: IC50 u opsegu od 5,26 μM do 83,42 μM; 5-LOX: IC50 u opsegu od 1,04 μM do 53,84 μM). Jedinjenja 4, 7, 8, 10 i 13 su se izdvojila kao 5-LOX inhibitori sa IC50 vrednostima u opsegu od 8,10 μM do 14,52 μM. Rezultati inhibicije COX-1 enzima ukazuju na to da su testirani dualni inhibitori slabi COX-1 inhibitori i da ispoljavaju zadovoljavajuću selektivnost prema COX-2 enzimu. Ispitana je antioksidativna aktivnost novosintetisanih jedinjenja. Dualni inhibitori 1, 2, 3 i 5 se ističu kao dobri potencijalni antioksidansi. U narednoj fazi istraživanja, razvijena je alternativna RP-HPLC metoda za procenu lipofilnosti jedinjenja. Najpogodniji RP-HPLC sistem se sastojao od C18 kolone (stacionarna faza), acetonitrila i fosfatnog pufera pH 7,4 i pH 5,5 u različitim odnosima (mobilna faza). Pasivna gastrointestinalna apsorpcija je procenjena primenom bioparticione micelarne hromatografije (BMC) i PAMPA testa. Na osnovu rezultata BMC testa, tj. dobijenih viših vrednosti retencionih faktora (k), izdvojena su jedinjenja sa potencijalno dobrom pasivnom gastrointestinalnom apsorpcijom (1, 2, 3, 5, 6, 7, 10 i 13). Rezultati PAMPA testa su bili poprilično usaglašeni sa rezultatima BMC testa i na osnovu dobijenih –logPe vrednosti očekuje se dobra pasivna gastrointestinalna apsorpcija u slučaju dualnih inhibitora 1, 2, 3, 5 i 6. Citotoksični efekti testiranih jedinjenja ispitani su primenom MTT testa na različitim kancerskim ćelijskim linijama (HCT 116, HT-29, BxPC-3). Dualni inhibitori 3, 5 i 6, kao i 5-LOX inhibitor 7, odabrani su za dalje eksperimente radiosenzitizacije (SRB test). Statistički značajni rezultati za kombinovani tretman u odnosu na samo zračenje zapaženi su na HCT 116 i BxPC-3 (3), BxPC-3 i HT-29 (6) i HT-29 (5 i 7). Inhibicija migracije SW620 ćelija je ispitana u testu zarastanja rana, a dualni inhibitori 1 i 3 su se istakli kao jedinjenja sa najboljim antimigratornim potencijalom. Jedinjenje 3 se izdvaja kao dualni inhibitor sa najboljim biološkim karakteristikama. Dual inhibition of COX-2 and 5-LOX enzymes represents a rational strategy for the development of more potent and safer anti-inflammatory drugs. It has been suggested that dual inhibitors could be useful in cancer therapy, as overexpression of COX-2 and 5-LOX enzymes in tumor cells has been observed. The first goal of research was the design of new, potential dual COX-2 and 5-LOX inhibitors using SAR analysis and computational methods (3D-QSAR study and molecular docking). COX-2 and 5-LOX inhibitory activity of newly synthesized and structurally characterized compounds was tested using commercially available COX-1, COX-2 and 5-LOX inhibitor screening assays. Compounds 1, 2, 3, 5, 6, 11 and 12 proved to be dual COX-2 and 5-LOX inhibitors. Obtained IC50 values were in the range of 5.26 - 83.42 μM (COX-2), and 1.04 - 53.84 μM (5-LOX). Compounds 4, 7, 8, 10 and 13 proved to be 5-LOX inhibitors (IC50 values were in the range of 8.10 - 14.52 μM). The tested dual inhibitors were weak COX-1 inhibitors (they showed selectivity towards COX-2 enzyme). The antioxidant activity of the newly synthesized compounds was tested and dual inhibitors 1, 2, 3 and 5 singled out as good potential antioxidants. In the next phase of research, an alternative RP-HPLC method was developed in order to assess the lipophilicity of compounds. The most suitable RP-HPLC system consisted of a C18 column (stationary phase), acetonitrile and phosphate buffer pH 7.4 and pH 5.5 in different ratios (mobile phase). Passive gastrointestinal absorption was assessed using biopartitioning micellar chromatography (BMC) and the PAMPA. Based on the obtained values of retention factors (k), it was concluded that the compounds 1, 2, 3, 5, 6, 7, 10 and 13 would have good pasive gastrointestinal absorption. The results of the PAMPA test were quite consistent with the results of the BMC test. Based on the obtained -logPe values, good passive gastrointestinal absorption is expected in the case of dual inhibitors 1, 2, 3, 5 and 6. Cytotoxic effects of the tested compounds on cancer cell lines (HCT 116, HT -29, BxPC-3) were examined using the MTT assay. Dual inhibitors 3, 5 and 6, as well as 5-LOX inhibitor 7, were selected for further radiosensitization experiments (SRB test). Statistically significant results for combined treatment versus radiation alone were observed in the case of HCT 116 and BxPC-3 (3), BxPC-3 and HT-29 (6) and HT-29 (5 and 7) cancer cell lines. Inhibition of SW620 cell migration was examined in the wound healing assay. Dual inhibitors 1 and 3 showed good antimigratory potential. Compound 3 singled out as a dual inhibitor with the best biological characteristics. Farmacija - Farmaceutsko-medicinska hemija / Pharmacy - Pharmaceutical-medicinal chemistry Datum odbrane: 16.10.2024. OX-2 i 5-LOX dualna inhibicija, antioksidansi, citostatici, antimigratorni agensi dual COX-2 and 5-LOX inhibition, antioxidants, cytostatics, antimigratory effect 615.31.015:577.1(043.3) 91552100 155878153 91552101 10288 2026-04-07T08:57:39.053Z 45 no 46 Jelena, 1993- Bošković 59378441 2024 63 mentor Olivera, 1964- Čudina 12653927 2024 63 mentor Vladimir, 1984- Dobričić 15607143 2024 63 član komisije Milkica, 1981- Crevar 22830183 2024 63 član komisije Jelena, 1969- Kotur-Stevuljević 12781671 2024 63 član komisije Nemanja, 1992- Đoković 28082279 2024 63 član komisije Otilija, 1983- Keta 116072969 2024 176 str. 13956078 http://phaidrabg.bg.ac.rs/o:38601 no yes 12 2026-04-07T08:57:39.320Z 70 11 1067094 1067100 COX-2 i 5-LOX dualna inhibicija, antioksidansi, citostatici, antimigratorni agensi dual COX-2 and 5-LOX inhibition, antioxidants, cytostatics, antimigratory effect 615.31.015:577.1(043.3) 1738 11A13 2024