This works describes synthesis, characterization and potential cytotoxity of O,O'- -dialkyl-(S,S)-ethylenediamine-N,N'-di-2-(4-methyl)-pentanoic acid, (S,S)-H4eddlCl2, as well as their corresponding ...palladium(II), platinum(II) and platinum(IV) complexes. Esters (S,S)-R2eddl•2HCl (R = ethyl, n-propyl, n-butyl and n-pentyl) were obtained by instillation of thionyl chloride in an appropriate absolute alcohol, and then refluxing with addition of (S,S)-ethylenediamine-N,N'-di-2-(4-methyl)-pentanoic acid. All of the esters were obtained as dihydrochlorides: (S,S)-Et2eddl•2HCl, (S,S)-Pr2eddl•2HCl, (S,S)-Bu2eddl•2HCl, (S,S)-Pe2eddl•2HCl. Esters were characterized by elemental analysis, IR and NMR spectroscopy, and in the case of (S,S)-Pr2eddl•2HCl the structure was confirmed by X-ray structural analysis. Complexes of palladium(II): PdCl2{(S,S)-Et2eddl}, PdCl2{(S,S)-Pr2eddl}, PdCl2{(S,S)-Bu2eddl} and PdCl2{(S,S)-Pe2eddl} were obtained by reaction of potassium- -tetrachloridopalladate(II) with synthesized esters and characterized by elemental analysis, IR and NMR spectroscopy. Complexes of platinum(II): PtCl2{(S,S)-Et2eddl}, PtCl2{(S,S)-Pr2eddl}, PtCl2{(S,S)-Bu2eddl} and PtCl2{(S,S)-Pe2eddl} were obtained by reaction of potassium- -tetrachloridoplatinate(II) with these esters and characterized by elemental analysis, IR and NMR spectroscopy. Complexes of platinum(IV): PtCl4{(S,S)-Et2eddl}, PtCl4{(S,S)-Pr2eddl}, PtCl4{(S,S)-Bu2eddl} and PtCl4{(S,S)-Pe2eddl} were obtained by reaction of potassium- -tetrachloridoplatinate(II) with mentioned esters and characterized by elemental analysis, IR and NMR spectroscopy. DFT calculations were performed for the synthesized complexes of platinum(II) and platinum(IV) and it was found that the (R,R)-N,N’-configuration isomer is with the lowest energy and formation of only this isomer should be expected, which is consistent with NMR spectroscopy. Cytotoxic activity of synthesized compounds was determined against chronic lymphocyte leukemia cells (CLL) and human cell lines: SW480 colorectal cancer, breast MDA-MB-361 and MDA-MB-453, Jurkat T leukemia and K562 chronic myelogenous leukemia compared with activity of cisplatin, as a reference anticancer drug. All compounds showed higher antitumoral activity then activity cisplatin against CLL calls. It was found that the most active complexes is complexes platinum(II) with n-Bu group in the ester chain. Complexes of the platinum(IV) have an exellent citotoxic activity in the inhibition of Jurkat and K562 cell lines, with is very similar to that shown by activity of cisplatin.
U ovoj doktorskoj disertaciji opisana je sinteza, karakterizacija i potencijalna citotoksičnost O,O'-dialkil estara (S,S)-etilendiamin-N,N'-di-2-(4- -metil)-pentanske kiseline, (S,S)-H4eddlCl2, kao i odgovarajućih paladijum(II), platina(II) i platina(IV) kompleksa. Estri (S,S)-R2eddl•2HCl (R = etil, n-propil, n-butil i n-pentil) su dobijeni ukapavanjem tionil-hlorida u odgovarajući apsolutni alkohol, a potom refluktovani uz dodatak (S,S)-etilendiamin-N,N'-di-2-(4-metil)-pentanske kiseline. Svi estri su dobijeni u obliku dihidrohlorida: (S,S)-Et2eddl•2HCl, (S,S)-Pr2eddl•2HCl, (S,S)-Bu2eddl•2HCl, (S,S)-Pe2eddl•2HCl. Estri su okarakterisani elementalnom analizom, IR i NMR spektroskopijom, a u slučaju (S,S)-Pr2eddl•2HCl struktura je potvrđena i rendgenskom strukturnom analizom. Kompleksi paladijuma(II): PdCl2{(S,S)-Et2eddl}, PdCl2{(S,S)-Pr2eddl}, PdCl2{(S,S)-Bu2eddl} i PdCl2{(S,S)-Pe2eddl} dobijeni su u reakciji kalijum- -tetrahloridopaladata(II) sa navedenim estrima i okarakterisani elementalnom analizom, IR i NMR spektroskopijom. Kompleksi platine(II): PtCl2{(S,S)-Et2eddl}, PtCl2{(S,S)-Pr2eddl}, PtCl2{(S,S)-Bu2eddl} i PtCl2{(S,S)-Pe2eddl} dobijeni su u reakciji kalijum- -tetrahloridoplatinata(II) sa navedenim estrima i okarakterisani elementalnom analizom, IR i NMR spektroskopijom. Kompleksi platine(IV): PtCl4{(S,S)-Et2eddl}, PtCl4{(S,S)-Pr2eddl}, PtCl4{(S,S)-Bu2eddl} i PtCl4{(S,S)-Pe2eddl} dobijeni su u reakciji kalijum- -heksahloridoplatinata(IV) sa navedenim estrima i okarakterisani elementalnom analizom, IR i NMR spektroskopijom. DFT proračuni rađeni su za sintetisane komplekse platine(II) i platine(IV) i ustanovljeno je da je (R,R)-N,N’-konfiguracioni izomer najniže energije, pri čemu je formiranje samo jednog izomera očekivano jer je u saglasnosti sa NMR spektroskopijom. Citotoksična aktivnost sintetisanih jedinjenja određena je na ćelijama hronične limfocitne leukemije (CLL) i humanim ćelijskim linijama: kolorektalnog karcinoma SW480, karcinoma dojki (MDA-MB-361 i MDA-MB-453), akutnoj T limfocitnoj leukemiji Jurkat i hroničnoj mijeloidnoj leukemiji K562 u poređenju sa aktivnošću cisplatine, kao referentnim antikancerogenim lekom. Sva jedinjenja pokazuju aktivnost veću od cisplatine prema CLL ćelijama. Nađeno je da najaktivnije jedinjenje kompleks platine(II) sa n-Bu grupom u estarskom lancu. Kompleksi platine(IV) imaju odličnu citotoksičnu aktivnost u inhibiciji Jurkat i K562 ćelijskih linija koja je veoma slična onoj koju pokazuje aktivnost cisplatine. Ključne reči: (S,S)-etilendiamin-N,N'-di-2-(4-metil)pentanska kiselina, (S,S)-R2eddl ligandi, R2edda ligandi, platina(IV) kompleksi, platina(II) kompleksi, paladijum(II) kompleksi, kristalna struktura, DFT proračuni, citotoksičnost
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.
The interest of European farmers for Vicia faba var. minor is increasing in the last years. This is mostly ascribing to the good nutritive value of faba beans and to the ban of use of genetically ...modified soybean and its byproducts in organic production. But Vicia faba contains some anti-nutritional factors that can also generate severe metabolic and digestive disorders in farm animals when utilized at high levels. The aim of the present paper is to summarize the structure and the synthesis pathways of these compounds and to review their biological effects. Because some of these factors are not destroyed by heating, the use of faba beans is strongly depending on animal species and level of utilization. In addition, the content of some anti-nutritional factors, e.g. glucosides, is variable depending on variety and environmental conditions, so that it is not easy to define sure levels of atoxicity. This is also more difficult in poultry, that are the most sensitive animals to the effects of glucosides
L'interesse degli agricoltori europei per il favino (Vicia faba var. minor) è in crescita negli ultimi anni. Questo fatto è da ricondurre soprattutto al buon valore nutritivo della sua granella e al divieto di utilizzazione della soia geneticamente modificata e dei suoi sottoprodotti nella produzione biologica. Ma il favino contiene alcuni fattori antinutrizionali che possono anche causare disturbi metabolici e digestivi negli animali in produzione zootecnica quando viene utilizzato a livelli elevati. Lo scopo di questo contributo consiste nel richiamare la struttura e le vie biosintetiche di questi composti e nell'esame dei loro effetti biologici. Dato che alcuni di questi fattori non vengono distrutti dal trattamento termico, l'impiego del favino dipende notevolmente dalla specie animale e dal livello di utilizzazione. Inoltre, il contenuto di alcuni fattori antinutrizionali, per esempio i glucosidi, varia in rapporto alla varietà e alle condizioni ambientali, per cui non risulta agevole stabilire livelli certi di atossicità. Questo è pure più difficile negli avicoli, che sono gli animali più sensibili agli effetti dei glucosidi.
In this study, we aimed to investigate the prognostic value of programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), and programmed cell death ligand 2 (PD-L2) expressions ...on immune and cancer cells in terms of survival in patients with lung adenocarcinoma, squamous cell carcinoma, and large cell carcinoma.
Between January 2000 and December 2012, a total of 191 patients (172 males, 19 females; mean age: 60.3±8.4 years; range, 38 to 78 years) who were diagnosed with non-small cell lung cancer and underwent anatomic resection and mediastinal lymph node dissection were retrospectively analyzed. The patients were evaluated in three groups including lung squamous cell carcinoma (n=61), adenocarcinoma (n=66), and large-cell carcinoma (n=64). The survival rates of all three groups were compared in terms of immunohistochemical expression levels of PD-1, PD-L1, and PD-L2.
The mean follow-up was 71.8±47.9 months. In all histological subtypes, PD-1 expressions on tumor and immune cells were observed in 33% (61/191) and in 53.1% (102/191) of the patients, respectively. Higher expression levels of PD-L1 and PD-L2 at any intensity on tumor and immune cells were defined only in lung adenocarcinomas, and PD-L1 and PD-L2 values were detected in 36.4% (22/64) of these patients. The PD-L1 expressions on tumor and immune cells were observed in 41.7% (10/24) and 25% (6/24) of the patients, respectively. The PD-L2 expressions on tumor and immune cells were detected in 16.7% (4/24) and 8.4% (2/24) of the patients, respectively. Univariate and multivariate analyses revealed that PD-1 expression in tumor cells was an independent prognostic factor in all histological subtypes.
Our study results suggest that PD-1 expression is a poor prognostic factor for overall survival in patients with completely resected adenocarcinoma, squamous cell carcinoma, and large cell carcinoma.
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