Synthesis, structure and properties of potentially biologically active propanoic acid derivatives

U cilju proučavanja potencijalne biološke aktivnosti derivata propanske kiseline u ovom radu su sintetisane dve serije jedinjenja. U prvoj seriji, sintetizovani su novi kompleksi prelaznih metala [Mn(II), Co(II), Ni(II), Cu(II), Zn(II)] sa oksaprozinom (Hoxa), nesteroidnim antiinflamatornim lekom. Lek i kompleksi okarakterisani su elementarnom i TG analizom, FT-IC, 1H NMR, 13C NMR, UV-Vis spektroskopijom i merenjem magnetne susceptibilnosti. Na osnovu elektronskih spektara i magnetnih momenata za sve komplekse utvrđena je (pseudo)oktaedarska geometrija. Sa izuzetkom Cu(II)-kompleksa, gde je ustanovljen mostovno bidentatni način vezivanja COO–-grupa, kod svih ostalih kompleksa na osnovu FT-IC spektara potvrđena je helatna koordinacija COO–-grupa. Opšta formula kompleksa je [M(H2O)2(oxa)2]∙xH2O, gde je x = 2 za M = Mn, Co i Ni i x = 1,5 za Zn. Binuklearni Cu(II)-kompleks, [Cu2(H2O)2(OH)(oxa)3]∙2H2O, ispoljava snažne Cu–Cu interakcije antiferomagnetnog tipa. Prvi put je ispitana in vitro antiproliferativna aktivnost sintetizovanih kompleksa prema ćelijskim linijama humanog karcinoma kolona (HCT-116), humanog karcinoma dojke (MDA-231), humanog karcinoma grlića materice (HeLa) i melanoma (Fem-x). Kod svih kompleksa uočen je statistički značajan antiproliferativni efekat. Zbog izuzetne aktivnosti, čak i pri nanomolarnim koncentracijama, Ni(II)-kompleks se pokazao kao potencijalni antiproliferativni agens. Drugu seriju jedinjenja čini grupa novih 2-(5-ariliden-2,4-dioksotetrahidrotiazol- 3-il)propanskih kiselina i odgovarajućih metil estara. Sva jedinjenja okarakterisana su temperaturom topljenja, elementarnom analizom, FT-IC, 1H NMR i 13C NMR spektroskopijom. Kristalna struktura metil-2-(5-(4-metoksifenil)metilen-2,4- -dioksotetrahidrotiazol-3-il)propionata određena je rendgenskom strukturnom analizom. Ispitivana je antiproliferativna aktivnost sintetisanih jedinjenja na ćelijskim linijama humanog karcinoma kolona (HCT-116), humanog karcinoma dojke (MDA-231) i mijeloidne leukemije (K-562). Rezultati ukazuju na bolju antiproliferativnost aktivnost sintetisanih metil estara u odnosu na odgovarajuće kiseline. Dodatno je ispitivana antimikrobna aktivnost, ali nije zabeležen njihov efekat naspram širokog spektra testiranih mikroorganizama. U nastavku istraživanja, urađena je elektrohemijska karakterizacija oksaprozina, cikličnom voltametrijom na elektrodi od zlata u 0,05 M rastvoru NaHCO3. Sintetizovan lek, njegov analitički standard i sadržaj aktivne supstance u tableti Duraprox® okarakterisan je jednom oksidacionom i tri redukcione reakcije. Uočena je linearna zavisnost anodnih struja od koncentracija leka na 0,83 V za analitički standard i na 0,85 V za tabletu Duraprox®. Elektroda od zlata je modifikovana adsorpcijom goveđeg serum albumina (bovine serum albumin, BSA) u 0,1 M fosfatnom puferu (pH = 7,4) i ispitana je koncentraciona zavisnost anodnih struja za analitički standard oksaprozina na BSA/Au elektrodi. Korišćenjem Lengmirove adsorpcione termodinamičke jednačine izračunata je konstanta vezivanja oksaprozina za BSA/Au elektrodu.In order to examine potentially biologically active propanoic acid derivatives two series of compounds have been synthesized. In the first series, novel transition metal [Mn(II), Co(II), Ni(II), Cu(II), Zn(II)] complexes with oxaprozin (Hoxa), a nonsteroidal anti-inflammatory drug, have been synthesized. The drug and complexes have been characterized by elemental and TG analysis, FT-IR, 1H NMR, 13C NMR, UV-Vis spectroscopy and magnetic susceptibility measurements. The (pseudo)octahedral geometry has been proposed for all complexes based on electronic spectra and magnetic moments. With exception of the Cu(II) complex, where bridging bidentate mode of COO– groups has been found, FT-IR spectra confirmed chelately coordinated COO– groups in the other complexes. The general formula of the complexes is [M(H2O)2(oxa)2]∙xH2O, with x = 2 for M = Mn, Co and Ni and x = 1.5 for Zn. The binuclear Cu(II) complex, [Cu2(H2O)2(OH)(oxa)3]∙2H2O, has strong Cu–Cu interactions of antiferromagnetic type. The complexes and Hoxa did not exhibit the cytotoxic effect to peritoneal macrophages. For the first time these complexes have been tested for their in vitro antiproliferative activity against human colon, breast, and cancer cell lines, HCT-116, MDA-231, HeLa and Fem-x, respectively. For all investigated compounds significant antiproliferative effects have been observed. Ni(II) complex has been shown to be a promising antiproliferative agent exerting excellent activity even in nanomolar concentrations. In the second series, novel 2-(5-arylidene-2,4-dioxotetrahydrothiazole-3- -yl)propanoic acids and six corresponding methyl esters have been synthesized. All compounds were characterized by melting points, elemental analysis, FT-IR, 1H and 13C NMR spectroscopy. Crystal structure of methyl-2-(5-(4-methoxybenzylidene)-2,4- -dioxotetrahydrothiazole-3-yl)propionate was confirmed by X-ray analysis. The antiproliferative activity of all synthesized compounds against human colon cancer, breast cancer and myelogenous leukemia cell lines, i.e. HCT-116, MDA-231 and K562, respectively, was evaluated. The results indicate that antiproliferative activity of the synthesized esters is better than the activity of the corresponding acids. Moreover, in vitro antimicrobial activity against a wide range of tested microorganisms was examined and not noticed. Further, electrochemical investigation of oxaprozin, using cyclic voltammetry on gold electrode in 0.05 M NaHCO3 was performed. The synthesized drug, its analytical standard and its content in Duraprox® tablets were characterized with one oxidation reaction and the three reduction reactions. All they exhibited the linear concentration dependency of anodic currents at 0.83 V for the analytical standard and 0.85 V for Duraprox® tablets. The strong adsorption of bovine serum albumin (BSA) on gold electrode in 0.1 M phosphate buffer solution (pH = 7.4) is shown and concentration dependency of anodic currents of oxaprozin standard on BSA/Au is studied. Following the Langmuir adsorption thermodynamic equation, the binding constants of oxaprozin on BSA/Au electrode was calculated