Computational methods for the description of pharmacologically relevant platinum complexes--molecular structure and bond dissociation.

Cancer is after cardiovascular disease the most frequent cause of death in Europe. In 28 of 53 countries considered in this area it is already the leading cause of death and expected to gain even more importance until the year 2020. Amongst the large arsenal of different anti-cancer drugs, platinum drugs belong to the first developed anticancer drugs and still have a large impact on cancer therapy. Nevertheless therapy with platinum-anticancer drugs is accompanied by severe adverse effects caused by frequent interactions with the amino acids of different human proteins. Computational chemistry offers methods to study such interactions and even those of not yet synthesized drugs in silico. For such studies a profound knowledge of the prediction quality of various computational methods towards platinum-drug-like complexes is necessary. By this article we are aiming on delivering important accuracy information of the frequently used computational methods. Most important findings are the high performance of the double hybrid functional B2PLYP for the calculation of geometries, even in small basis sets, followed by BP86 and PBE and the still acceptable performance of the semi-empirical Method PM6-D3H4X for extremely large systems. To follow absolute energies of the dissociation process, LPNO-CEPA and B3LYP-D3 can be suggested while SCS-MP2 shows an extremely narrow standard deviation and a low maximum error, which make it an ideal candidate for relative energy calculations in the exploration of reaction mechanisms.

Multiparametric characterization of nonelectroactive self-assembled monolayers during their formation.

The formation of nonelectroactive self-assembled monolayers (SAMs) at the electrode/electrolyte interface was characterized with simultaneous impedance, gravimetric, and direct current measurements. In the presence of specifically adsorbing inorganic ions, this provides key information about the formation of SAMs. Gravimetric measurements allow an estimation of the adsorbate surface coverage; and completion of the assembly process can then be monitored in real-time. Electrochemical impedance spectroscopy measurements play a multifunctional role: they enable elucidation of the physical models of the interface, provide the information about the effective capacitance of SAMs thus probing the dielectric properties of the adsorbed layers, and evaluate the ability of charged electrolyte components to approach the electrode surface through the SAM (using adsorbing/desorbing SO4(2-) as an electroactive probe). The latter is important to assess the extent of defects in the formed organic layers. Finally, monitoring the direct current during SAM formation together with the collected gravimetric data can give additional important information about the process. A series of n-mercaptoalcohols with different hydrocarbon chain length adsorbing at Au electrodes was used as the model object to evaluate the proposed approach.

Cell response of Escherichia coli to cisplatin-induced stress.

Cisplatin is undoubtedly one of the most common and successful anticancer drugs worldwide. Though its DNA-based mechanism of action is well established, the contribution of the proteome to this process remains unclear. The possible impact of particular Escherichia coli proteins on the cytostatic activity of cisplatin was the subject of this study. Our main focus was not only the "bottom-up" identification of novel cisplatin protein targets through LC/LC-MS/MS analysis, but also a label-free quantification of their regulation profile by spectral-counting. The regulation of two proteins, aconitate hydratase 2 and 60 kDa chaperonin 1, could be linked to a platinated amino acid in the protein sequence, whereas in the cases of 30S ribosomal protein S1 and enolase, it could be shown that cisplatin fragments are coordinated to an essential site for the functionality of the protein. Nucleoside triphosphate pyrophosphohydrolase (MazG) regulates the programmed cell death and was found to be platinated on the protein surface, which probably correlates with the established mode of action. A possible new chapter in the understanding of cisplatin's mechanism of action and its severe side effects is opened, since evidence is provided that platinated proteins are not only involved in cellular stress response but also in energy metabolism through glycolysis and catabolic processes, in gene regulatory mechanisms and protein synthesis.

Sulfur-substituted naphthalimides as photoactivatable anticancer agents: DNA interaction, fluorescence imaging, and phototoxic effects in cultured tumor cells.

A series of sulfur-substituted naphthalimides (1-5) was prepared and investigated as antitumor drugs. Initial DNA interaction studies (by the fluorescence quenching method, UV/vis and CD spectroscopy, thermal denaturation, topoisomerase Western blot analysis, and DNA photocleavage experiments) expectedly suggested the DNA and topoisomerase as main targets of the agents. Fluorescence spectroscopic and microscopic experiments indicated a significant sensitivity of the emission intensities of 3 and 5 to the cellular environment and confirmed the cellular uptake and biodistribution into cell compartments for 1-3 and 5. A comparative evaluation of the antiproliferative effects under different experimental setups (concerning drug exposure period and an additional short-time UV irradiation) revealed significant phototoxic effects for the environmentally sensitive compounds 3 and 5 and strongly suggested the further development of sulfur-substituted naphthalimides for potential use in photodynamic tumor therapy.

DNA Intercalators for Detection of DNA Hybridisation: SCS(MI)-MP2 Calculations and Electrochemical Impedance Spectroscopy.

Quantum mechanical SCS(MI)-MP2/cc-pVTZ calculations predict the strength of proflavine, ellipticine and 1-pyrenemethylamine intercalation into single-stranded (ss) and double-stranded (ds) DNA. The results were compared with experimental results obtained from electrochemical impedance spectroscopy (EIS). Similar interaction energies of ellipticine with the guanine-cytosine base pair compared to the individual nucleobases guanine and cytosine suggested non-specific binding also to ssDNA. Accordingly, EIS identified ellipticine as being non-selective and therefore unsuitable for the detection of DNA hybridisation. The interaction energy of proflavine is significantly higher than the minimum required energy for a single intercalation site, and substantially lower with respect to the minimum energy needed for binding with ssDNA. In EIS studies, proflavine did not show any change in the charge-transfer resistance with respect to ssDNA and a decrease with respect to dsDNA. Calculations showed that 1-pyrenemethylamine has sufficiently high interaction energy to intercalate into dsDNA, however, the interaction energy towards ssDNA is close to the minimum required value, suggesting a weak interaction with ssDNA. EIS measurements support the calculations. A method for the calculation of interaction energies is provided, which can be used to characterise the interaction strength between new intercalators and DNA before being synthesised.

Tetra-cationic imidazoliumyl-substituted phosphorus-sulfur heterocycles from a cationic organophosphorus sulfide.

The reaction of imidazoliumyl-substituted P((III)) cations of type [L((R,Me))PCl2](+) (3a,b(+); L(R,Me) = imidazolium-2-yl a: R = Me; b: R = iPr) with (Me3Si)2S leads to the formation of tetra-cationic, eight-membered phosphorus sulfur heterocycles [L((R,Me))PS]4(4+) (9a,b(4+)), which can be explained by the tetramerization of the intermediately formed cationic phosphorus monosulfide [L((R,Me))PS](+) (8a,b(+)). The P4S4 ring adopts a crown conformation as observed for cyclo-S8. The Lewis base DMAP (4-dimethylaminopyridine) initiates a deoligomerization- and dismutation reaction of 9a,b(4+) to give P((I)) centered cation [L((R,Me))2P](+) (12a,b(+)) and phosphorus disulfide [(DMAP)2PS2](+) (14(+)).

Alkynyl gold(I) phosphane complexes: Evaluation of structure-activity-relationships for the phosphane ligands, effects on key signaling proteins and preliminary in-vivo studies with a nanoformulated complex.

Gold alkynyl complexes with phosphane ligands of the type (alkynyl)Au(I)(phosphane) represent a group of bioorganometallics, which has only recently been evaluated biologically in more detail. Structure-activity-relationship studies regarding the residues of the phosphane ligand (P(Ph)3, P(2-furyl)3, P(DAPTA)3, P(PTA)3, P(Et)3, P(Me)3) of complexes with an 4-ethynylanisole alkyne ligand revealed no strong differences concerning cytotoxicity. However, a relevant preference for the heteroatom free alkyl/aryl residues concerning inhibition of the target enzyme thioredoxin reductase was evident. Complex 1 with the triphenylphosphane ligand was selected for further studies, in which clear effects on cell morphology were monitored by time-lapse microscopy. Effects on cellular signaling were determined by ELISA microarrays and showed a significant induction of the phosphorylation of ERK1 (extracellular signal related kinase 1), ERK2 and HSP27 (heat shock protein 27) in HT-29 cells. Application of 1 in-vivo in a mouse xenograft model was found to be challenging due to the low solubility of the complex and required a formulation strategy based on a peanut oil nanoemulsion.

Characterisation of Complex Electrode Processes using Simultaneous Impedance Spectroscopy and Electrochemical Nanogravimetric Measurements.

The methodology and illustrative examples of application are presented for a technique that simultaneously combines electrochemical impedance spectroscopy (EIS) and nanogravimetric measurements; the latter are implemented using a so-called electrochemical quartz crystal nanobalance (EQCN). The combination of EIS and EQCN provides a powerful method for the characterisation of many complex processes at electrochemical interfaces. This method gives in one relatively simple experiment more detailed information than is available from conventional electrochemical techniques. The combined measurements can be performed either as a function of time, at a constant electrode potential, or under potentiodynamic conditions, as a function of the electrode potential. Herein, we show how this can be applied to enable more accurate investigation of processes that occur at boundaries between electrodes and electrolytes. The application examples range from eletrocatalysis, in which evaluation of a catalyst is performed simultaneously with its formation, and the intercalation and electrodeposition of thin metal films to in situ characterisation of non-electroactive self-assembled monolayers during their formation.

Highly cytotoxic substitutionally inert rhodium(III) tris(chelate) complexes: DNA binding modes and biological impact on human cancer cells.

The antiproliferative properties and cellular impact of novel substitutionally inert rhodium(III) complexes of the types [Rh{(CH3)2 NCS2}2(pp)]Cl 3-5 (pp=5,6-Me2phen, dpq, dppz) and OC-6-23-[Rh(2-S-py)2(pp)]Cl 6 and 7 (2-S-py=pyridine-2-thiolate; pp=dpq, dppz) have been investigated for the adherent human cancer cell lines MCF-7 and HT-29 and for non-adherent Jurkat cells. Whereas CD and viscosity measurements indicate that the polypyridyl ligands of 4 and 5 intercalate into CT DNA, this is not the case for the analogous pyridine-2-thiolate complexes 6 and 7. Complexes 3-7 all exhibit a high antiproliferative activity towards MCF-7 and HT-29 cells, with IC(50) values in the range 0.055-0.285 µM. As established by online monitoring with a cell-based sensor chip, the highly cytostatic complex 6 (IC(50)=0.059 and 0.078 µM) invokes an immediate concentration-dependent reduction of MCF-7 cell respiration and a time-delayed decrease in cellular impedance, which can be ascribed to the induction of cell death. Annexin V/PI assays demonstrated that 6 also has a pronounced antiproliferative activity towards Jurkat cells and that it invokes extensive apoptosis and high concentrations of reactive oxygen species in these leukemia cells. The observation of a dose-dependent inhibition of the oxygen consumption of isolated mice mitochondria indicates the involvement of an intrinsic mitochondrial pathway in this process.

Comparative in vitro evaluation of N-heterocyclic carbene gold(I) complexes of the benzimidazolylidene type.

Gold(I) complexes with a 1,3-diethylbenzimidazol-2-ylidene N-heterocyclic carbene (NHC) ligand of the type NHC-Au-L (L=-Cl, -NHC, or -PPh3) were comparatively evaluated as thioredoxin reductase (TrxR) inhibitors and antimitochondrial anticancer agents. Different effects were noted in various biochemical assays (e.g., inhibition of TrxR, cellular and mitochondrial uptake, or effects on mitochondrial membrane potential), and this was related to properties of the complexes such as bond dissociation energies and overall charge. Remarkable antiproliferative effects, a strong induction of apoptosis, and enhancement of reactive oxygen species (ROS) formation as well as other effects on tumor cell metabolism confirmed the promising potential of the complexes as novel anticancer chemotherapeutics.