Predrag Tepeš

LC―NMR and LC―MS identification of an impurity in a novel antifungal drug icofungipen

Successful use of LC-NMR and LC-MS for rapid identification of an impurity in a novel antifungal drug icofungipen has been demonstrated. Complementary information obtained from the two methods made it possible to determine the structure of A1 prior to its isolation and purification. Stop-flow LC-NMR ((1)H and DQFCOSY), LC-MS and LC-MS/MS spectra have shown that A1 is structurally related to icofungipen. It was later isolated and prepared synthetically and its structure was corroborated by high-resolution NMR spectroscopy.

Free and bound state structures of 6-O-methyl homoerythromycins and epitope mapping of their interactions with ribosomes.

The solution and solid state conformations of several 6-O-methyl homoerythromycins 1-4 were studied using a combination of X-ray crystallography, NMR spectroscopy and molecular modelling calculations. In the solid state 1 was found to exist as the two independent molecules with similar structures termed 3-endo-folded-out. In solution a significant conformational flexibility was noticed especially in the C2 to C5 region. The compounds 1 and 2 unlike 14-membered macrolides adopted the 3-endo-folded-out conformation while 3 and 4 existed in the classical folded-out conformation. TrNOESY and STD experiments showed that 1 and 2 bound to the Escherichia coli ribosome while 3 and 4, lacking the cladinose sugar, did not exhibit binding activities, this being in accordance with biochemical data. The bound conformations were found to be very similar to the free ones, some small differences were observed and discussed. The STD experiments provided evidence on binding epitopes. The structural parts of 1 and 2 in close contact with ribosome were similar, however the degree of saturation transfer was higher for 2. The differences between tr-NOE data and STD enhancements in 1 and 2 arouse as a consequence of structural changes upon binding and a closer proximity of 2 to the ribosome surface. An understanding of the molecular mechanisms involved in the interaction of macrolides with ribosomes can help in developing strategies aiming at design of potential inhibitors.

New Dinuclear Molybdenum(V) Complexes With β′‐Hydroxy‐β‐enaminones Containing a 4‐Hydroxy‐2‐pyrone Ring

New dinuclear molybdenum(V) complexes of the general formula [Mo2O4L2D2], were prepared by the reaction of [Mo2O3(acac)4] (acac = acetylacetonate ion) with β′-hydroxy-β-enaminones (L). All prepared complexes consist of Mo2O42+ cores coordinated by two ligands L via two donor oxygen atoms as in the analogous β-diketonates. The usual octahedral coordination around the molybdenum atoms is completed by the monodentate solvent molecules D (methanol, ethanol, or 2-propanol). All compounds were characterized by elemental analyses, IR, one- and two-dimensional NMR spectra, and thermal analysis, and some of them by X-ray crystallography (1a, 4a, 8b, and 9a). (© Wiley-VCH Verlag GmbH, 69451 Weinheim, Germany, 2002)

Conformational analysis of oleandomycin and its 8-methylene-9-oxime derivative by NMR and molecular modelling

Conformations of the 14-membered macrolide antibiotic oleandomycin and its 8-methylene-9-oxime derivative were determined in various solvents. The experimental NMR data--coupling constants and NOE contacts--were compared with the results of molecular modelling--molecular mechanics calculations and molecular dynamics simulations. The conformational changes, on the right-hand side of the 14-membered ring, affected mostly the 3JH2,H3 values and NOE crosspeaks H3 or H4 to H11. Oleandomycin was found to be present predominantly in the C3-C5 folded-in conformations in DMSO-d6 solution, whereas in buffered D2O, acetone-d6 and CDCl3, there was a mixture of folded-in and folded-out conformational families. The predominant conformation of the 8-methylene-oleandomycin-9-oxime derivative in solution was a folded-out one although different amounts of folded-in conformation were also present depending on the solvent. Oleandrose and desosamine sugar moieties adopted the usual and expected chair conformation. The conformation around the glycosidic bonds, governing the relative orientation of sugars vs. the lactone ring, showed a certain flexibility within two conformationally close families. We believe that by combining the experimental NMR data and the molecular modelling techniques, as reported in this paper, we have made significant progress in understanding the conformational behaviour and properties of macrolides. Our belief is based on our own current studies on oleandomycins as well as on the previously reported results and best practices concerning other macrolides. A rational for macrolide conformational studies and advances in methodology has been suggested accordingly.

Efficiency of subsurface flow constructed wetland with trickling filter

Effective wastewater purification in subsurface flow constructed wetlands must include adequate pretreatment and ensure a sufficient amount of dissolved oxygen. In a pilot-scale operation, a subsurface flow constructed wetland (CW) consisted of a primary settlement tank, a trickling filter for pretreatment and two serially assembled basins. The trickling filter was added to ensure sufficient aeration, increase purification of the wastewater and shorten the wastewater purification time. The estimated nominal flow was 0.7 m3/d. The experiments were conducted using the wastewater from the municipal sewage canal of the city of Zagreb, with utilization of three different flows: 0.72 (A), 1.44 (B) and 2.88 (C) m3/d. The efficiency of the purification process was monitored over a period of three years (TSS, BOD5, COD, NH4-N, NO2-N, PO4-P, dissolved oxygen, temperature and pH). The experimental results showed an increase in the removal efficiency with a doubling of the nominal flow from 0.7 to 1.44 m3/d, which could be related to the implementation of the trickling filter where high removal rates were achieved.