RESUMO
The trithiolato bridged diruthenium complex DiRu-1 [(p-MeC6H4iPr)2Ru2(SC6H4-p-But)3]+ is highly cytotoxic against various cancer cell lines, but its exact mode of action remains unknown. The present 1H HR-MAS NMR-based metabolomic study was performed on ovarian cancer cell line A2780, on its cis-Pt resistant variant A2780cisR, and on the cell line HEK-293 treated with 0.03 µM and 0.015 µM of DiRu-1 corresponding to full and half IC50 doses, respectively, to investigate the mode of action of this ruthenium complex. The resulting changes in the metabolic profile of the cell lines were studied using HR-MAS NMR of cell lysates and a subsequent statistical analysis. We show that DiRu-1 in a 0.03 µM dose has significant impact on the levels of a number of metabolites, such as glutamine, glutamate, glutathione, cysteine, lipid, creatine, lactate, and acetate, especially pronounced in the A2780cisR cell line. The IC50/2 dose shows some significant changes, but full IC50 appears to be necessary to observe the full effect. Overall, the metabolic changes observed suggest that redox homeostasis, the Warburg effect, and the lipid metabolism are affected by DiRu-1.
RESUMO
(1)H high resolution magic angle spinning (HR-MAS) NMR spectroscopy was applied in combination with multivariate statistical analyses to study the metabolic response of whole cells to the treatment with a hexacationic ruthenium metallaprism [1](6+) as potential anticancer drug. Human ovarian cancer cells (A2780), the corresponding cisplatin resistant cells (A2780cisR), and human embryonic kidney cells (HEK-293) were each incubated for 24 h and 72 h with [1](6+) and compared to untreated cells. Different responses were obtained depending on the cell type and incubation time. Most pronounced changes were found for lipids, choline containing compounds, glutamate and glutathione, nucleotide sugars, lactate, and some amino acids. Possible contributions of these metabolites to physiologic processes are discussed. The time-dependent metabolic response patterns suggest that A2780 cells on one hand and HEK-293 cells and A2780cisR cells on the other hand may follow different cell death pathways and exist in different temporal stages thereof.
Assuntos
Antineoplásicos/farmacologia , Metaboloma/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Compostos Organometálicos/farmacologia , Rutênio/farmacologia , Antineoplásicos/química , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Células HEK293 , Humanos , Espectroscopia de Ressonância Magnética , Neoplasias/patologia , Compostos Organometálicos/química , Rutênio/químicaRESUMO
The reactivity of three hexacationic arene ruthenium metallaprisms towards isolated nucleotides and a short DNA strand was investigated using NMR spectroscopy, ESI mass spectrometry, UV/Vis and circular dichroism spectroscopy. The metallaprism built from oxalato-bridging ligands reacts rapidly in the presence of deoxyguanosine monophosphate (dGMP) and deoxyadenosine monophosphate, while the benzoquinonato derivative only reacts with dGMP. On the other hand, the larger metallaprism incorporating naphtoquinonato bridges remains stable in the presence of nucleotides. The reactivity of the three hexacationic metallaprisms with the decameric oligonucleotide d(CGCGATCGCG)2 was also investigated. Analysis of the NMR, MS, UV/Vis and CD data suggests that no adducts are formed between the oligonucleotide and the metallaprisms, but electrostatic interactions, leading to partial unwinding of the double-stranded oligonucleotide, were evidenced.
Assuntos
Antineoplásicos/química , Complexos de Coordenação/química , Nucleotídeos/química , Oligodesoxirribonucleotídeos/química , Rutênio/químicaRESUMO
Interactions between three hexacationic arene ruthenium metallaprisms, [(p-cymene)6Ru6(tpt)2(dhnq)3](6+), [(p-cymene)6Ru6(tpt)2(dhbq)3](6+) and [(p-cymene)6Ru6(tpt)2(oxa)3](6+), and a series of human proteins including human serum albumin, transferrin, cytochrome c, myoglobin and ubiquitin have been studied using NMR spectroscopy, mass spectrometry and circular dichroism spectroscopy. All data suggest that no covalent adducts are formed between the proteins and the metallaprisms. Indeed, in most cases electrostatic interactions, leading to precipitation of protein-metallaprism aggregates, have been observed. In addition, with the smallest proteins, ubiquitin, myoglobin and cytochrome c, the presence of the hexacationic arene ruthenium metallaprisms induces structural changes of the proteins, as emphasized by circular dichroism. The results suggest that proteins are certainly a biological target for these metalla-assemblies.
Assuntos
Complexos de Coordenação/química , Citocromos c/química , Monoterpenos/química , Mioglobina/química , Rutênio/química , Ubiquitina/química , Precipitação Química , Dicroísmo Circular , Complexos de Coordenação/síntese química , Humanos , Modelos Moleculares , Ressonância Magnética Nuclear Biomolecular , Conformação Proteica , Albumina Sérica/química , Soluções , Eletricidade Estática , Transferrina/químicaRESUMO
The crystal structure of chlorido{µ-2-[(2-oxidobenzylidene)amino]ethanolato-κ(4)O,N,O':O'}{2-[(2-oxidobenzylidene)amino]ethanolato-κ(3)O,N,O'}trivinylditin(IV), [Sn2(C2H3)3(C9H9NO2)2Cl], is disordered above 178â K. A doubling of the unit-cell volume is observed on cooling. The asymmetric unit at 93â K contains two ordered molecules. The phase transition corresponds to an order-disorder transition of one vinyl group bound to the Sn(IV) atom.
Assuntos
Complexos de Coordenação/química , Estanho/química , Cristalografia por Raios X , Ligantes , Modelos Moleculares , Transição de FaseRESUMO
NMR spectroscopy has proved extremely beneficial in the investigation of inorganic drugs from the time that cisplatin was first introduced into the clinic more than 30 years ago. Both (195)Pt and (15)N NMR were used in early studies and made a major contribution in the understanding of the molecular mechanism of action from model studies involving reactions with amino acids and nucleotides. Over the past decade, ruthenium drugs have proved to be a valuable alternative to platinum drugs, and NMR has also provided unique insights into their molecular mechanism of action including investigations of simple aquation reactions, protein binding and the kinetics and sequence selectivity of DNA binding interactions. In this article, emphasis is given to define the cellular targets and elucidate some of the mechanistic profiles of recent ruthenium-based organometallic compounds offering efficacy toward cancer cells, by various NMR techniques.
Assuntos
Complexos de Coordenação/farmacologia , Espectroscopia de Ressonância Magnética/métodos , Rutênio/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Complexos de Coordenação/química , Humanos , Rutênio/químicaRESUMO
Reactions between the cationic triangular metallaprism [(p-cymene)(6)Ru(6)(tpt)(2)(dhnq)(3)](6+) ([1](6+)) [tpt is 2,4,6-tri(pyridine-4-yl)-1,3,5-triazine; dhnq is 5,8-dihydroxy-1,4-naphthoquinonato] and Arg, His, Lys, ascorbic acid, lactic acid and glutathione (GSH) have been studied at 37 °C in aqueous solution at pD 7 using NMR spectroscopy and electrospray ionisation mass spectrometry. Coordination to the imidazole nitrogen atom of His or to the basic NH/NH(2) groups in Arg and Lys slowly displaces the dhnq and tpt ligands from the (p-cymene)Ru units, and subsequently additional coordination to the amino and carboxylato groups forms stable N,N,O metallacycles. Compared with our previously reported study with the analogous metallaprism [(p-cymene)(6)Ru(6)(tpt)(2)(dhbq)(3)](6+) ([2](6+)) (dhbq is 2,5-dihydroxy-1,4-benzoquinonato), the larger metallaprism [1](6+) appears to be significantly more stable, and disassembled in the presence of Arg, His and Lys after only 12 h of incubation. Moreover, the reaction with His is not complete, since only 14 % of His reacted after more than 1 week of incubation. Solutions of [1](6+) are also able to catalyse oxidation of the thiol group of Cys and GSH to give the corresponding disulfides and of ascorbic acid to give the corresponding dehydroascorbic acid. However, the results are markedly different from those obtained with metallaprism [2](6+): the oxidation of Cys and ascorbic acid is not complete, and the formation of intermediate adducts could be evidenced. On the other hand, the oxidation of GSH remains fast and is completed after only 12 h. Oxidation of GSH to give the corresponding disulfide may explain its higher in vitro anticancer activity as compared with [2](6+). Our results suggest that metallaprism [1](6+) is more robust than [2](6+), may remain intact in the bloodstream and, therefore, may enter cancer cells undamaged, thus confirming the drug delivery potential for such water-soluble organometallic cages.
Assuntos
Aminoácidos/química , Complexos de Coordenação/química , Espectroscopia de Ressonância Magnética , Rutênio/química , Espectrometria de Massas por Ionização por Electrospray , Ácido Ascórbico/química , Estabilidade de Medicamentos , Ácido Láctico/química , Ligantes , Estrutura MolecularRESUMO
The relative affinity of the cationic triangular metallaprism, [(pCH(3)C(6)H(4)Pr(i))(6)Ru(6)(tpt)(2)(dhbq)(3)](6+) ([1](6+)), for various amino acids, ascorbic acid, and glutathione (GSH) has been studied at 37 °C in aqueous solutions at pD 7, using NMR spectroscopy and electrospray ionization mass spectrometry (ESI-MS). The metallaprism [1](6+), which is constituted of six (pCH(3)C(6)H(4)Pr(i))Ru corners bridged by three 1,4-benzoquinonato (dhbq) ligands and connected by two 2,4,6tri(pyridin4yl)1,3,5-triazine (tpt) triangular panels, disassembled in the presence of Arg, His, and Lys, while it remains intact with Met. Coordination to the imidazole nitrogen atom in His or to the basic NH/NH(2) groups in Arg and Lys displaces the dhbq and tpt ligands from the (p-cymene)Ru units, and subsequent coordination to the amino and carboxylato groups forms stable N,N,O metallacycles. The binding to amino acids proceeds rapidly, as determined by NMR spectroscopy. Interestingly, solutions of [1](6+) are able to catalyze oxidation of the thiol group of Cys and GSH to give the corresponding disulfides and of ascorbic acid to give the corresponding dehydroascorbic acid. Competition experiments with Arg, Cys, His, and Lys show the simultaneous formation of one single adduct, the (p-cymene)Ru-His complex, and oxidation of Cys to cystine. Furthermore, the (p-cymene)Ru-His complex formed upon the addition of His to [1][CF(3)SO(3)](6) is able to oxidize Cys to cystine much more efficiently than [1](6+). These results provide evidence against interaction with proteins as process in the release of encapsulated guest molecules. Oxidation of Cys and GSH to give the corresponding disulfides may explain the in vitro anticancer activity of [1](6+).