Metallocenyl 7-ACA Conjugates: Antibacterial Activity Studies and Atomic-Resolution X-ray Crystal Structure with CTX-M β-Lactamase

The conjugation of organometallic groups to current β-lactam antibiotics is a field of increasing study due to the ability of certain organometallic groups to enhance the antibiotic potency of these drugs. Herein, we report the antibacterial properties of two metallocenyl (ferrocenyl and ruthenocenyl) 7-aminocephalosporanic acid (7-ACA) antibiotic conjugates. Continuing a trend we found in our previous studies, the ruthenocenyl conjugate showed greater antibacterial activity than its ferrocenyl counterpart. Compared with the previously published 7-aminodesacetoxycephalosporanic acid (7-ADCA) conjugates, the 3-acetyloxymethyl group significantly improved the compounds’ activity. Furthermore, the Rc-7-ACA compound was more active against clinical Staphylococcus aureus isolates than the ampicillin reference. Noticeably, neither of the two new compounds showed an undesirable toxic effect in HeLa and L929 cells at the concentrations at which they displayed strong antibacterial effects. The antibacterial activity of the two metallocenyl 7-ACA derivatives was further confirmed by scanning electron microscopy (SEM). SEM micrographs showed that bacteria treated with metallocenyl 7-ACA derivatives feature cell wall damage and morphology changes. Using a CTX-M-14 β-lactamase competition assay based on nitrocefin hydrolysis, we showed that the Rc-7-ACA bound more favorably to CTX-M-14 than its ferrocenyl counterpart, again confirming the superiority of the ruthenocenyl moiety over the ferrocenyl one in interacting with proteins. We also report a 1.47 Å resolution crystal structure of Rc-7-ACA in complex with the CTX-M-14 E166A mutant, an enzyme sharing a similar active site configuration with penicillin-binding proteins, the molecular target of β-lactam antibiotics. These results strengthen the case for the antibacterial utility of the Rc and Fc groups.     

Design of novel FN3 domains with high stability by a consensus sequence approach

The use of consensus design to produce stable proteins has been applied to numerous structures and classes of proteins. Here, we describe the engineering of novel FN3 domains from two different proteins, namely human fibronectin and human tenascin-C, as potential alternative scaffold biotherapeutics. The resulting FN3 domains were found to be robustly expressed in Escherichia coli, soluble and highly stable, with melting temperatures of 89 and 78°C, respectively. X-ray crystallography was used to confirm that the consensus approach led to a structure consistent with the FN3 design despite having only low-sequence identity to natural FN3 domains. The ability of the Tenascin consensus domain to withstand mutations in the loop regions connecting the β-strands was investigated using alanine scanning mutagenesis demonstrating the potential for randomization in these regions. Finally, rational design was used to produce point mutations that significantly increase the stability of one of the consensus domains. Together our data suggest that consensus FN3 domains have potential utility as alternative scaffold therapeutics.     

Towards Heterogeneous and Green Versions of Os Dihydroxylation Catalysis

Os-catalyzed dihydroxylation will find more widespread application if truly heterogeneous catalysts are available which use O₂ or H₂O₂ as terminal oxidants. This paper discusses and critically evaluates the concepts by which Os can be anchored on solid supports. New methods are also available for reoxidation of hexavalent to octavalent Os.     

Heterogeneous Enzyme Mimics Based on Zeolites and Layered Hydroxides

Careful combination of a metal compound,a ligand and an inorganic support material leads to supramolecular catalysts that mimic the structural, organizational and functional aspects of enzyme activity.After discussing essential features of metalloenzyme-catalyzed reactions and coordination chemistry in inorganic hosts, we present examples of supramolecular materials selected from our own work that eventually resulted in useful catalysts for organic transformations in the liquid phase.     

Foam processing of poly(ethylene-co-vinyl acetate) rubber using supercritical carbon dioxide

Soft rubber foams like poly(ethylene-co-vinyl acetate) (EVA) are industrially applied in a broad range of products, including sports gear, insulation materials and drug delivery systems. In contrast to glassy polymers, few studies in literature concern the foaming of soft rubbers using supercritical carbon dioxide. In this study, open microporous matrices of EVA have been formed with CO₂. Prior to the foam expansion, sorption and swelling isotherms of CO₂ in EVA have been measured and the obtained isotherms have been correlated using the Sanchez-Lacombe equation of state. Additionally, a pressure-independent diffusion coefficient of CO₂ in EVA has been obtained from these experiments. The microporous foams have been formed by a pressure quench of the CO₂-swollen polymer matrix. Sorption pressure as well as temperature and decompression times appear to determine the pore size and bulk density of the foam. These parameters allow for a control of the foam structure of EVA rubbers.     

Hydrotalcite-like anionic clays in catalytic organic reactions

Hydrotalcite-like anionic clays (HTs; or layered double hydroxides, LDHs) can be used as precursors to mixed oxides, but their catalytic potential reaches much further. This review demonstrates that HTs can be successfully applied in a broad spectrum of organic reactions, with advantages such as improved activity, selectivity, metal dispersion, less waste production, and an improved recuperation of immobilized catalysts. HTs can be used in the as-synthesized form or after different pretreatments. The HT as such is a solid base or, depending on the elemental composition of its octahedral layers, may have redox properties. Specific metals can be incorporated either as a cation in the octahedral layer or as an anion via exchange. The anion-exchange strategy is especially successful in the context of heterogenizing homogeneous catalysts. A particular advantage for base catalysis is that the number and strength of the basic sites can be tuned precisely to a specific reaction. HTs are excellent materials to design bifunctional redox-base catalysts or to control the acid-base properties around a heterogenized metal complex. Potential applications of HTs range from the production of large-scale basic chemicals to the synthesis of small-scale specialty chemicals.