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

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: [(η6-p-cymene)Ru(L1–3)Cl2], where L1–3 is 3-acetylpyridine (1), 4-acetylpyridine (2) and 2-amino-5-chloropyridine (3), correspondingly, [(η6-p-cymene)Ru(HL4,5)Cl2], where HL4 i HL5 are respectively isonicotinic acid (4) and nicotinic acid (5) and [(η6-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 [(η6-pcymene) RuCl(L11)], where HL11 is picolinic acid (11). Complex [(η6-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. Knowing that great number of ruthenium complexes although devoid of direct cytotoxicity in vitro, exhibit important antimetastatic characteristics, potential of our Ru