Sinteza, karakterizacija i ispitivanje biološke aktivnosti kompleksa bakra(II) sa ß-aminokiselinama

Abstract

Uvod: Bakar(II)-jon je zbog prelazno tvrdo-mekih karakteristika idealan za koordinovanje sa različitim donorskim atomima, a zbog esencijalne uloge koju ima u organizmu njegovi kompleksi su potencijalno manje toksični. Aminokiseline preko atoma azota i kiseonika lako formiraju koordinativno-kovalentne veze sa prelaznim metalima. Zbog biološke aktivnosti i farmakološkog značaja, β-aminokiseline su pogodni ligandi u sintezi potencijalnog leka. Materijal i metode: U okviru ove doktorske disertacije opisani su postupci sinteza β-aminokiselina i odgovarajućih kompleksa bakra(II) sa β-aminokiselinama. Strukture sintetisanih liganada potvrđene su na osnovu infracrvene (IR) i nuklearno- -magnetno-rezonancione spektroskopije (1N i 13S NMR), dok su sastav i strukture sintetisanih kompleksnih jedinjenja pretpostavljene na osnovu rezultata elementalne mikroanalize, infracrvene i elektronsko-paramagnetno-rezonancione spektroskopije (EPR). Stvarna struktura kompleksa bakar(II)-jona i 2-(1-aminocikloheksil)sirćetne kiseline potvrđena je i na osnovu rezultata rendgenske strukturne analize. Interakcije sintetisanih kompleksa sa molekulom DNK utvrđene su na osnovu kinetičkih merenja, apsorpciono spektroskopskih merenja, fluorescentnih merenja i merenja viskoziteta DNK rastvora. Broj i vijabilnost tumorskih ćelija (4T1, CT26, LLC1) nakon primene rastvora kompleksnih jedinjenja određena je kolorimetrijskim MTT testom. Antimikrobna aktivnost sintetisanih jedinjenja ispitana je mikrodilucionom metodom. Rezultati: Potvrđena je kvadratno-planarna struktura sinetisanih kompleksa. Vrednosti konstanti vezivanja, smanjenja intenziteta fluorescencije i povećanje relativnog viskoziteta rastvora DNK ukazuju na interakciju kompleksa i molekula DNK. Od ispitivanih ćelijskih linija najveću osetljivost prema sintetisanim kompleksima pokazale su ćelije karcinoma pluća, LLC1. Oba kompleksa indukuju apoptozu ćelija karcinoma kolona (ST26) i pri nižim koncentracijama imaju bolju aktivnost od cisplatine. Ispitivane supstance ispoljavaju nisku i selektivnu antimikrobnu aktivnost koja je verovatno posledica njihovih slabih lipofilnih karakteristika. Zaključak: Sinteza kompleksa bakra(II) sa različitim ligandima daje mogućnost razvoja potencijalnih lekova. Ispitivanje biološke aktivnosti svakog potencijalnog farmakoterapeutika omogućava da se približnije poveže struktura kompleksa sa mehanizmom dejstva i antitumorskim i antimikrobnim potencijalom.

Introduction: Copper(II)-ion is ideal for coordinating with different donor atoms due to its soft-hard characteristics while transferring, and because of its essential role in organism, its complexes are potentially less toxic. Amino acids, through the atoms of nitrogen and oxygen, easily form coordinate-covalent bonds with transition metals. β-amino acids are suitable ligands in the synthesis of a potential medicine because of their biological activity and pharmacological significance. Materials and methods: This PhD thesis describes the syntheses procedures of β-amino acids and corresponding copper(II)-complexes with β-amino acids. The structures of synthesized ligands were confirmed based on infrared (IR) and nuclear-magnetic-resonance spectroscopy (1Н and 13С NMR), while the composition and the structures of synthesized complex compounds were assumed by the results of elemental microanalysis, infrared and electron paramagnetic resonance (EPR). The real structure of copper(II)-ion complex and 2-(1-aminocyclohexyl) acetic acid was confirmed from the results of Roentgen structural analysis. The interactions of synthesized complexes with the DNA molecule were determined by kinetic measurements, absorption spectroscopic measurements, fluorescent measurements and the measurements of the DNA solution viscosity. The number and the viability of tumor cells (4T1, CT26, LLC1), after applying complex compounds solutions, was determined by МТТ colorimetric technique. Antimicrobial activity of synthesized compounds was examined using microdilution method. Results: Square planar structure of synthesized compounds was confirmed. The values of binding constants, fluorescence intensity reduction and the increase of relative DNA solution viscosity indicated the interaction between complexes and the DNA molecule. From the tested cell lines, lung cancer cells (LLC1) showed the largest sensitivity towards synthesized complexes. Both complexes induced cell apoptosis of colon cancer (СТ26) and had higher activity in lower concentrations than cisplatin. The tested substances expressed low and selective antimicrobial activity which was probably the result of their weak lipophilic characteristics. Conclusion: The synthesis of copper(II)-complexes with different ligands offers the possibility for the development of potential medicines. Biological activity examination of each potential drug enables more approximate connection of the structure of complexes with the mechanism of action and anticancer and antimicrobial potentials.