Investigation of antiproliferative and in vitro antimetastatic potential of picolinate ruthenium(II)-cymene complex : comparison to a series of ruthenium(II)-arene complexes with similar structure

Cilj ove disertacije bio je ispitivanje potencijala antiproliferativnog i in vitro antimetastatskog dejstva serije novosintetisanih rutenijum(II)-arenskih kompleksa. U pitanju je serija Ru(II)-arenskih kompleksa sledeih strukturnih formula: [(6-pcimen) Ru(L1–3)Cl2], gde je L1–3: 3-acetilpiridin (1), 4-acetilpiridin (2) i 2-amino-5- hloropiridin (3), kao i [(6-p-cimen)Ru(HL4,5)Cl2], gde HL4 i HL5 odovara izonikotinskoj kiselini (4) i nikotinskoj kiselini (5) i [(6-p-cimen)Ru(HL6–9)Cl], gde H2L6–9 predstavlja 2,3-piridindikarboksilnu kiselinu (6), 2,4-piridindikarboksilnu kiselinu (7), 2,5-piridindikarboksilnu kiselinu (8) i 2,6- piridindikarboksilnu kiselinu (9), i [(6-p-cimen)RuCl(L11)], gde je HL11 pikolinska kiselina (11). Kompleks (10) je polazni kompleks [(6-p-cymene)2RuCl2]2 koji je korišen za sinteze navedenih kompleksa u reakciji sa odgovarajuim ligandima. Antiproliferativna aktivnost Ru(II)- arenskih kompleksa je ispitana na: šest tumorskih elijskih linija (HeLa, MDA-MB-361, MDA-MB-453, FemX, B16, LS-174), na dve transformisane endotelijalne linije (EA.hy 926, MS1) i na jednoj normalnoj humanoj liniji (MRC-5). Za dalja ispitivanja poreenja povezanosti strukture i aktivnosti odabrana su dva kompleksa sa monodentatno vezanim piridinskim ligandom (1 i 3) i dva kompleksa sa bidentano vezanim piridinskim ligandom (6 i 7), koji nisu imali znaajnu citotoksinu aktivnost i pikolinato rutenijum(II)-cimenski kompleks (11), kao kompleks sa znaajnom aktivnošu. Potencijal ispitivanih kompleksa da indukuju promene na nivou elijskog ciklusa odreen je korišenjem protonog citometra nakon bojenja tretiranih elija sa propidijum-jodidom. Takoe, korišenjem testa za detekciju rane faze apoptoze,dvokolornim bojenjem elija sa aneksinom i propidijum-jodidom i analize na protonom citometru ispitan je potencijal kompleksa 11 da indukuje apoptozu. Distribucija rutenijuma(II) u proteinskoj i DNK frakciji HeLa elija tretiranih sa ispitivanim kompleksima utvrena je korišenjem indukovano kuplovane plazme sa optiko emisionom spektrometrijom (ICP-OES)...The aim of this thesis was to investigate antiproliferative and in vitro antimetastatic potential of series of newly synthesized ruthenium(II)-arene complexes. It is a series of Ru(II)-arene complexes of general formula: [(6-p-cymene)Ru(L1–3)Cl2], where L1–3 is 3-acetylpyridine (1), 4-acetylpyridine (2) and 2-amino-5-chloropyridine (3), correspondingly, [(6-p-cymene)Ru(HL4,5)Cl2], where HL4 i HL5 are respectively isonicotinic acid (4) and nicotinic acid (5) and [(6-p-cymene)Ru(HL6–9)Cl], where H2L6–9 represent 2,3-pyridinedicarboxylic acid (6), 2,4-pyridinedicarboxylic acid (7), 2,5-pyridinedicarboxylic acid (8) and 2,6- pyridinedicarboxylic acid (9), and [(6-pcymene) RuCl(L11)], where HL11 is picolinic acid (11). Complex [(6-pcymene) 2RuCl2]2 (10) was starting complex used for the synthesis of complexes of this series with the corresponding ligands. Analysis of cell growth inhibition caused by Ru(II)-arene complexes was performed on: six tumor cell lines (HeLa, MDA-MB-361, MDA-MB-453, FemX, B16, LS-174), on two transformed endothelial lines (EA.hy 926, MS1) and on one normal human cell line (MRC-5). For further examination of comparison of structure and activity we used two complexes with monodentate bonded pyridine ligand (1 an 3) and two with bidentate bonded pyridine ligand (6 and 7), which didn´t have any important cytotoxic activity and picolinate ruthenium(II)-cymene complex (11), as complex with important activity. Potential of investigated complexes to induce cell cycle perturbations was determined after staining of treated cells with propidium iodide (PI) on flow cytometer. As well as determination of induction of early apoptotic changes by complex 11, after two colors staining with Annexin V-FITC and PI and analysis on flow cytometer. Ru(II) distribution among the DNA and protein fractions in HeLa cells treated with investigated complexes was determined using inductive coupled plasma with optical emissione spectrometry (ICP-OES). In this study we evaluated whether DNA-repair-dependent signaling, as a result of interaction with DNA, which includes components of NER or MMR is utilized in cell response to ruthenium(II)-p-cymene complexes, by following expression of ERCC1 (mRNA and protein level) and MSH2 (protein level) using Quantitative Real-Time PCR (RQ-PCR) and Western blot..

Supplementary data for the article: Nikolic, S.; Rangasamy, L.; Gligorijevic, N.; Arandelovic, S.; Radulovic, S.; Gasser, G.; Grguric-Sipka, S. Synthesis, Characterization and Biological Evaluation of Novel Ru(II)-Arene Complexes Containing Intercalating Ligands. J. Inorg. Biochem. 2016, 160, 156–165. https://doi.org/10.1016/j.jinorgbio.2016.01.005

Supplementary material for: [https://doi.org/10.1016/j.jinorgbio.2016.01.005]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2272]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3638

Rendgenska strukturna analiza i citotoksična aktivnost pikolinato rutenijum(II)-arenskog kompleksa

A ruthenium(II)-arene complex with picolinic acid, [(eta(6)-p-cymene)RuCl(pico)]center dot H(2)O, was prepared by the reaction of [(eta(6)-p-cymene)RuCl(2)](2) with picolinic acid in a 1:2 molar ratio in 2-propanol. The compound was characterized by elemental analysis, and IR and NMR spectroscopy. X-ray diffraction analysis showed that the molecule adopts a "three-leg piano-stool" geometry, which is common for this type of complexes. The cytotoxic activity of the complex was tested in two human cancer cell lines HeLa (cervix) and FemX (melanoma) by MTT assay. The IC(50) values were at 82.0 and 36.2 mu mol dm(-3) for HeLa and FemX cells, respectively.Rutenijum(II)-arenski kompleks sa pikolinskom kiselinom [(η6-p-cimen)RuCl(pikolinato)]·H2O sintetisan je u reakciji [(η6-p-cimen)RuCl2]2 kompleksa sa pikolinskom kiselinom u molskom odnosu 1:2 u izopropanolu. Jedinjenje je okarakterisano elementalnom analizom, IC i NMR spektroskopijom. Analiza difrakcijom X-zracima pokazala je da molekul ima tzv. 'three-leg piano-stool' geometriju koja je karakteristična za ovaj tip kompleksa. Citotoksična aktivnost kompleksa je određena na dve humane tumorske ćelijske linije, HeLa (grlića materice) i FemX (melanoma), MTT testom. IC50 vrednosti su bile 82,0 i 36,2 µmol dm-3 za HeLa i FemX ćelije, redom

Antitumor activity of organoruthenium complexes with chelate aromatic ligands, derived from 1,10-phenantroline: Synthesis and biological activity

The monocationic chloro complexes containing chelating N∩N ligands: [(η6-p-cymene)Ru(L1–4)Cl]+ (1–4), where L1 = 4-methyl-1,10-phenantroline, L2 = dipyrido[3,2-a:2′,3′-c]phenazine, L3 = 11-chloro-dipyrido[3,2-a:2′,3′-c]phenazine, L4 = 11-nitro-dipyrido[3,2-a:2′,3′-c]phenazine; p-cymene = 1-methyl-4-isopropylbenzene) have been prepared and characterized as the hexafluorophosphate salts. The biological activity of 1–4 has been investigated in selected 2D monolayer cell cultures (A549, PANC-1, MDA-MB-231, MRC-5). All investigated ruthenium complexes showed similar or even better cytotoxicity to cisplatin. However, there was no significant reduction in growth of PANC-1 cells in a 3D cell culture of multicellular tumor spheroids (MCTS) after treatment with 2–4, while the cisplatin treatment induced retardation in MCTS growth. Flow cytometry analysis of the cell cycle of PANC-1 cells shows that 3 caused changes of cell cycle phase distribution characterized by slight accumulation of cells in the G2-M phase. Absence of the Sub-G1 phase in the cell cycle of the treated cells indicated that there was no fragmentation of DNA for the analyzed time intervals (48 and 72 h treatment). Fluorescent microscopy, after acridine orange/ethidium bromide staining, revealed that the investigated ruthenium complexes induced some characteristics of apoptotic morphology (shrinking and condensation of chromatin) with notably preserved integrity of the plasma membrane. Investigation of cellular uptake and DNA - fraction accumulation performed by inductively coupled plasma mass spectrometry in PANC-1 cells with equimolar concentrations (5 μM) of 2–4 and cisplatin showed more efficient cellular uptake and DNA - fraction accumulation of complex 3 compared to complexes 2 and 4

Supplementary data for article: Milenković, M. R.; Bacchi, A.; Cantoni, G.; Radulović, S. S.; Gligorijević, N.; Aranđelović, S.; Sladić, D.; Vujčić, M.; Mitić, D.; Anđelković, K. K. Synthesis, Characterisation and Biological Activity of Co(III) Complex with the Condensation Product of 2-(Diphenylphosphino)Benzaldehyde and Ethyl Carbazate. Inorganica Chimica Acta 2013, 395, 33–43. https://doi.org/10.1016/j.ica.2012.09.043

Supplementary material for: [https://doi.org/10.1016/j.ica.2012.09.043]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1618

Supplementary data for article: Marković, V.; Erić, S.; Stanojković, T.; Gligorijević, N.; Aranđelović, S.; Todorović, N.; Trifunović, S. S.; Manojlović, N.; Jelic, R.; Joksović, M. D. Antiproliferative Activity and QSAR Studies of a Series of New 4-Aminomethylidene Derivatives of Some Pyrazol-5-Ones. Bioorganic and Medicinal Chemistry Letters 2011, 21 (15), 4416–4421. https://doi.org/10.1016/j.bmcl.2011.06.025

Supplementary material for: [https://doi.org/10.1016/j.bmcl.2011.06.025]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1175

Supplementary data for the article: Pavlović, M.; Tadić, A.; Gligorijević, N.; Poljarević, J.; Petrović, T.; Dojčinović, B.; Savić, A.; Radulović, S.; Grgurić-Šipka, S.; Aranđelović, S. Synthesis, Chemical Characterization, PARP Inhibition, DNA Binding and Cellular Uptake of Novel Ruthenium(II)-Arene Complexes Bearing Benzamide Derivatives in Human Breast Cancer Cells. Journal of Inorganic Biochemistry 2020, 210, 111155. https://doi.org/10.1016/j.jinorgbio.2020.111155

Supplementary material for: [https://doi.org/10.1016/j.jinorgbio.2020.111155]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/4048

Supplementary data for article: Baroud, A. A.; Mihajlović-Lalić, L. E.; Gligorijević, N.; Aranđelović, S.; Stanković, D.; Radulović, S.; Van Hecke, K.; Savić, A.; Grgurić-Šipka, S. Ruthenium(II) Bipyridine Complexes: From Synthesis and Crystal Structures to Electrochemical and Cytotoxicity Investigation. Journal of Coordination Chemistry 2017, 70 (5), 831–847. https://doi.org/10.1080/00958972.2017.1282611

Supplementary material for: [https://doi.org/10.1080/00958972.2017.1282611]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/2423

Supplementary material for the article: Čobeljić, B.; Milenković, M.; Pevec, A.; Turel, I.; Vujčić, M.; Janović, B.; Gligorijević, N.; Sladić, D.; Radulović, S.; Jovanović, K.; et al. Investigation of Antitumor Potential of Ni(II) Complexes with Tridentate PNO Acylhydrazones of 2-(Diphenylphosphino)Benzaldehyde and Monodentate Pseudohalides. Journal of Biological Inorganic Chemistry 2016, 21 (2), 145–162. https://doi.org/10.1007/s00775-015-1315-x

Supplementary material for: [https://doi.org/10.1007/s00775-015-1315-x]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1906