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  • Radić Gordana

    06/2012
    Dissertation

    The discovery of anticancer activity of cis-diammindichloridoplatinum(II)complexes, better known as cisplatin (cis-DDP) and related platinum complexes has stimuled the interest for obtaining more efficient complexes of other metals and ligands. Some the first complexes used in clinical treatments against tumors were palladium(II)complexes as analogues of cisplatin. Although the first results were not encouraging,palladium complexes have been studied to longer extent. Palladium(II) complexes show less antitumor activity than corresponding platinum complexes due to their high instability. In order to overcome these problems, many studies have been directed toward the use of chelating ligands which would reduce the reactivity of palladium(II) complexes. So far a small number of complex compounds of palladium(II) have been synthesized with O,O'-dialkyl esters edda-type ligands as bidentate ligands, as well as S-alkyl derivatives of thiosalicylic acid. Therefore, this dissertation describes the synthesis and characterization of some edda-type ligands, their esters, S-alkyl derivatives of thiosalicylic acid and corresponding palladium(II) complexes with obtained ligands. The synthesis and characterization of some ligands and corresponding palladium(II) complexes are described in this PhD thesis. Firstly, the synthesis and characterization of O,O'-dialkyl esters of ethylenediamine-N,N'-di-(S,S)-2-propanoic acid H2-(S,S)-eddp is described. These compounds were obtained by direct reaction between tetradentate ligand and the corresponding absolute alcohol (ethanol, 1-propanol, 1-butanol, 1-pentanol) in a molar ratio 1:2 with the addition of gaseous hydrogen chloride. As described above, O,O'-dialkyl esters of ethylenediamine-N,N'-di-(S,S)- -2-(3-methyl)-butanoic acid, H2-(S,S)-eddv were synthesized was also done. The synthesis of S-alkyl derivatives of thiosalicylic acid. Bidentate S,O donor ligands were obtained by direct reaction between the sodium salt of thiosalicylic acid and corresponding alkyl halides (methyl, ethyl, propyl, butyl, benzyl) in a molar ratio 1:1. The synthesis of the tetradentate ligand meso-1,2-diphenyl-ethylenediamine-N,N'-di-3- -propanoic acid is shown at the end of this dissertation. Structure and composition of the isolated ligands were assumed using by elemental microanalysis, IR, 1 H and 13C NMR spectroscopy, and confirmed by X-ray diffraction analysis in the case of meso-1,2- -diphenyl-ethylenediamine-N,N'-di-3-propanoic acid monohydrate dicholorohydrate, H2-1,2-dpheddp•2HCl•H2O. The obtained ligands were used for synthesis of the corresponding palladium(II) complexes. The composition of obtained square-planar palladium(II) complexes was confirmed by elemental microanalysis. The structure of isolated complexes is assumed on basis of infrared, 1 H and 13C NMR spectroscopy and confirmed on basis of X-ray diffraction analysis in the case of chlorido(S,S)-ethylenediamine-N- -(O-ethyl-2-(3-methyl)butanoate)-N'-2-(3-methyl)-butanoate-palladium(II) complex, PdCl(et-(S,S)-eddv) and bis-(S-benzyl-thiosalycilato)palladium(II) complex, Pd(S-bz-thiosal)2. Antimicrobial activity of synthesized ligands and corresponding palladium(II) complexes was tested by microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concetration (MMC) were determined in this process. Some complexes showed significant antifungal activity on pathogenic fungi Aspergillus flavus and Aspergillus fumingatus. However, synthesized palladium(II) complexes showed limited antibacterial activity. In vitro antitumor activity of the complexes cis-PdCl2(1,2-dpheddp) and s-cis-PtCl2(1,2-dpheddp) was determined on the tumor cells of the 4T1 and B16F1, and the number of cells was determined after 72 hours by using MTT technique. The study found that both compounds showed lower antitumor activity compared to cisplatin. Otkriće antitumorskih osobina cis-diamindihloridoplatina(II)kompleksa, poznatijeg pod imenom cisplatina (cis-DDP) i sličnih kompleksa platine povećalo je interesovanje za dobijanje još efikasnijih kompleksa drugih metala i liganada. Među prvim kompleksima korišćenim u kliničkim ispitivanjima protiv tumora bili su paladijum(II) analozi cisplatine. Iako prvobitni rezultati nisu bili ohrabrujući, kompleksi paladijuma(II) su mnogo šire proučavani. Paladijum(II) kompleksi su skoro uvek pokazivali manju antitumorsku aktivnost u odnosu na odgovarajuće komplekse platine zbog njihove velike labilnosti. Da bi se ovi problemi prevazišli, mnoga istraživanja su bila usmerena ka upotrebi helatnih liganada koji bi umanjili reaktivnost paladijum(II) kompleksa. Do sada je sintetisan mali broj kompleksnih jedinjenja paladijuma(II) sa O,O'-dialkil estrima liganada edda-tipa kao bidentatnim ligandima, kao i sa S-alkil derivatima tiosalicilne kiseline. Stoga je u okviru ove Doktorske disertacije opisana sinteza i karakterizacija nekih liganada edda-tipa, njihovih estara, kao i S-alkil derivata tiosalicilne kiseline i odgovarajućih kompleksa paladijuma(II). U ovoj Doktorskoj disertaciji opisana je sinteza i karakterizacija većeg broja liganada i odgovarajućih kompleksa paladijuma(II). Prvo je opisana sinteza i karakterizacija O,O'-dialkil estara etilendiamin-N,N'-di-(S,S)-2-propanske kiseline (H2-(S,S)-eddp). Ova jedinjenja su dobijena direktnom reakcijom između tetradentatnog liganda i odgovarajućeg apsolutnog alkohola (etanol, 1-propanol, 1-butanol, 1-pentanol) u molskom odnosu 1:2, uz uvođenje gasovitog hlorovodonika. Na prethodno opisan način sintetisani su i O,O'-dialkil estri etilendiamin- N,N'-di-(S,S)-2-(3-metil)-butanske kiseline (H2-(S,S)-eddv). Takođe, prikazana je i sinteza S-alkil derivata tiosalicilne kiseline. Pomenuti bidentatni S,O donorski ligandi dobijeni su direktnom reakcijom između natrijumove soli tiosalicilne kiseline i odgovarajućih alkil halogenida (metil-, etil-, propil-, butil- i benzil) u molskom odnosu 1:1. Na kraju prikazana je i sinteza tetradentatnog liganda meso-1,2-difenil-etilendiamin-N,N'-di-3-propanske kiseline. Strukture i sastav izolovanih liganada pretpostavljene su primenom elementalne mikroanalize, infracrvene, 1 H i 13C NMR spektroskopije, a potvrđene na bazi rezultata rendgenske strukturne analize u slučaju: meso-1,2- -difenil-etilendiamin-N,N′-di-3-propanske kiseline dihlorhidrata monohidrata, (H2-1,2-dpheddp•2HCl•H2O). Dobijeni ligandi su upotrebljeni za sintezu odgovarajućih kompleksa paladijuma(II). Sastav dobijenih kvadratno-planarnih kompleksa paladijuma(II) potvrđen je elementalnom mikroanalizom. Struktura izolovanih kompleksa pretpostavljena je na osnovu infracrvene, 1 H i 13C NMR spektroskopije, a potvrđena je na bazi rendgenske strukturne analize u slučaju hlorido((S,S)-etilendiamin-N-(O-etil-2-(3-metil)-butanoat)-N'-2-(3- -metil)-butanoatopaladijum(II) kompleksa, PdCl(et-(S,S)-eddv) i bis-(S-benzil- -tiosalicilato)paladijum(II) kompleksa, Pd(S-bz-thiosal)2. In vitro antimikrobna aktivnost sintetisanih liganada i odgovarajućih kompleksa paladijuma(II) testirana je mikrodilucionom metodom. Tom prilikom određene su minimalne inhibitorne (MIK) i minimalne mikrobicidne koncentracije (MMK) nagrađenih jedinjenja. Pojedini kompleksi su pokazali značajnu antifungalnu aktivnost na patogenim gljivama Aspergillus flavus i Aspergillus fumigatus. Međutim, sintetisani paladijum(II) kompleksi su pokazali umerenu antibakterijsku aktivnost. In vitro antitumorska aktivnost kompleksa cis-PdCl2(1,2-dpheddp) kao i s-cis-PtCl2(1,2-dpheddp), ispitivana je na ćelijama tumora 4T1 i B16F1, a broj ćelija određivan nakon je 72 sata korišćenjem MTT tehnike. Ispitivanjem je utvrđeno da oba jedinjenja pokazuju nižu antitumorsku aktivnost u poređenju sa cisplatinom.