Unusual Ar-H/Rh-H J(HH) NMR coupling in complexes of rhodium(III): experimental evidence and theoretical support for an eta1-arene structure

The synthesis and structural properties of three new hydridorhodium(III) complexes are reported. Hydrogenolysis of the cyclometalated rhodium dichloride complexes [RhCl(2)[(S,S)-benbox(Me(2))]] (2a-c) leads to formation of the new complexes [RhCl(2)(H)[(S,S)-ip-benbox(Me(2))H]] (3a-c) in 45% to 85% yield. Compounds 3a-c were found to have unusual features by NMR spectroscopy: in particular, downfield shifted aryl proton resonances (8.88-9.03 ppm) that were coupled to the rhodium hydride resonances. Using X-ray crystallographic studies, a variety of solid- and solution-state characterization techniques, and DFT calculations, these features were attributed to the presence of weak pi-type eta(1)-arene interactions in 3a-c.     

An automated, sheathless capillary electrophoresis-mass spectrometry platform for discovery of biomarkers in human serum

A capillary electrophoresis-mass spectrometry (CE-MS) method has been developed to perform routine, automated analysis of low-molecular-weight peptides in human serum. The method incorporates transient isotachophoresis for in-line preconcentration and a sheathless electrospray interface. To evaluate the performance of the method and demonstrate the utility of the approach, an experiment was designed in which peptides were added to sera from individuals at each of two different concentrations, artificially creating two groups of samples. The CE-MS data from the serum samples were divided into separate training and test sets. A pattern-recognition/feature-selection algorithm based on support vector machines was used to select the mass-to-charge (m/z) values from the training set data that distinguished the two groups of samples from each other. The added peptides were identified correctly as the distinguishing features, and pattern recognition based on these peptides was used to assign each sample in the independent test set to its respective group. A twofold difference in peptide concentration could be detected with statistical significance (p-value < 0.0001). The accuracy of the assignment was 95%, demonstrating the utility of this technique for the discovery of patterns of biomarkers in serum.     

Quantitative comparison of molecular electrostatic potentials for structure-activity studies

An approach for representing, efficiently calculating and comparing discrete three-dimensional molecular electrostatic potentials using a quantitative similarity index (MEP-SI) based on a Carbo-type formalism is presented. A radial-type (MACRA) grid representation is described that provides more efficient storage of MEP information than a cubic grid of similar range, appropriate emphasis, and a convenient means for restricting the comparison of MEP functions to a local molecule region. The MACRA based MEP-SI formalism was used to evaluate the suitability of a variety of approximate methods for efficiently calculating the MEP for use in MEP-SI comparison of dissimilar molecules. The Mulliken charge method was found inadequate, while the method of potential-derived charges (PDCs), with additional charges for lone electron pairs included on sulfur, provided an efficient and sufficiently accurate representation of the MEP for this purpose. Convergence of the MEP-SI with respect to MACRA grid extent and mesh size was demonstrated; the effect of MEP error and different grid point emphasis in the MACRA versus the cubic grid results was investigated, and MEP-SI results were compared for different forms of the SI equation. The methodology proposed in this study provides an efficient and practical means for comparing MEP functions for two molecules and gives discriminating results for a sample series of molecular analogues consistent with expectations.     

PHOTODIMERIZATION OF 1,3-DIMETHYL-5-CHLOROURACIL*

Abstract— 1,3-Dimethyl-5-chlorouracil dimerizes upon direct photolysis in aqueous solution to give one major photodimer. X-Ray analysis has been used to identify the dimer as the anti head-to-tail isomer, IV.     

Probing the interaction of HTI-286 with tubulin using a stilbene analogue

HTI-286 is a synthetic analogue of the natural product hemiasterlin. HTI-286 is a potent antitumor agent that induces tubulin oligomerization. To investigate the binding stoichiometry and the binding site during this ligand-induced tubulin association, we synthesized an analogue of HTI-286 containing the chromophore stilbene. Using the distinct absorbance of the stilbene analogue, we determined the amounts of inhibitors bound to different tubulin oligomers by analytical ultracentrifugation. Herein we describe our findings based on these experiments. At the ratio of inhibitor to protein equal to or greater than 1, the stilbene analogue induces oligomerization of tubulin to a ring structure. The binding stoichiometry in the ring is one inhibitor per tubulin monomer (defined as an alpha/beta-heterodimer). At the ratio of inhibitor to protein less than 1, tubulin forms multiple intermediates, with the binding stoichiometry less than one inhibitor per tubulin monomer for all intermediates. The stable complex between the inhibitor and tubulin monomer was not detected under our experimental conditions. The binding site of the stilbene analogue does not overlap with the classic tubulin-binding agent, colchicine.     

Reflections on the calculation and publication of potentiometrically-determined formation constants

Calculation and reporting procedures are described, which aim to improve the quality and comparability of published formation constant values obtained by glass-electrode potentiometry. Ways in which the processing of data by computer optimization programs can be standardized are the main concern, particularly in respect of improving the usefulness of equilibrium data through incorporation into large publically available databases. These recommendations may be particularly valuable to researchers beginning in this field as well as to those who determine formation constants only occasionally.     

Simultaneous determination of dopa and carbidopa enantiomers by capillary zone electrophoresis

Dopa and carbidopa, components of the dual therapy for Parkinson's disease treatment, are both provided as single enantiomers, since their D-forms are inactive. To ensure the efficiency and safety of the therapy, these D-enantiomers, therefore, should be considered as impurities. In this paper, the enantioseparation power of different types of cyclodextrins, both neutral and charged ones, on dopa and carbidopa enantiomers was tested. Three methods of simultaneous separation of dopa and carbidopa enantiomers were developed, using highly sulfated beta-cyclodextrin and sulfated beta-cyclodextrin as chiral selector, in normal and reversed polarity mode. Two methods among these three were found sensitive enough for the quantitation of 0.1% D-enantiomers in L-forms (impurity level). After the optimization study, the best method was selected, using 16 mM sulfated beta-cyclodextrin in 15 mM sodium phosphate buffer pH 2.45, an uncoated fused-silica capillary (50 num inner diameter, 30 cm total length), and an applied voltage of -12 kV. This method is robust and efficient, with very high resolution for all peaks within a short analysis time of 10 min. Quantitatively, the method offers a limit of detection (LOD) of 0.2 nug/mL and a limit of quantitation (LOQ) of 0.5 nug/mL for both D-dopa and D-carbidopa, which is equivalent to 0.02% and 0.05% against the respective L-enantiomers. A linear relationship was found between the concentration of the analyte and the corresponding peak area in a range of 0.5-2.0 nug/mL.     

Use of SDS micelles to stabilize a ternary intermediate in the reaction of ferrioxamine B and 1,10-phenanthroline

Spectrophotometric measurements of the reaction of ferrioxamine B (FeHDFB(+)) with 1,10-phenanthroline (phen) reveal the presence of a ternary intermediate complex in both aqueous solution and an aqueous solution of 0.16 M sodium dodecyl sulfate (SDS). The stoichiometry of the intermediate is Fe(H(2)DFB)(phen)(2+) on the basis of a Schwarzenbach analysis of spectrophotometric data obtained at variable pH and phen concentrations. The ternary complex formation constant for the reaction FeHDFB(+) + H(+) + phen right arrow over left arrow Fe(H(2)DFB)(phen)(2+) is log K = 6.96 in aqueous solution and log K = 8.64 in aqueous 0.16 M SDS. The enhanced stability of Fe(H(2)DFB)(phen)(2+) in micellar solution was analyzed in terms of the pseudophase ion-exchange (PPIE) model of micellar reactions. The association constants for the binding of each reactant to the micellar pseudophase were measured by ultrafiltration. According to PPIE model calculations, the enhanced stability of Fe(H(2)DFB)(phen)(2+) in micellar SDS arises from a proximity effect created by the high local concentrations of reactants in the micellar pseudophase. The calculations also indicate that an inhibitory medium or compartmentalization effect is operative since the observed micellar enhancement is much smaller than predicted by the PPIE model. The micellar stabilization of the Fe(H(2)DFB)(phen)(2+) intermediate and the overall conversion of FeHDFB(+) to Fe(phen)(3)(2+) are discussed as a possible model system for siderophore iron release in microbial organisms.