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ABSTRACT: A small series of 5-(hetero)aryl-modified nucleoside phosphonates was synthesized via an 8-step procedure including a Wittig reaction and Suzuki-Miyaura coupling. An unanticipated anomerization during phosphonate deprotection allowed us to isolate both anomers of the 5-substituted 2'-deoxy-uridine phosphonates and assess their antiviral activity against a broad panel of viruses.
Nucleosides Nucleotides & Nucleic Acids 03/2012; 31(3):256-72. · 0.90 Impact Factor
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ABSTRACT: We explored the influence of modifications of uridine 5'-methylenephosphonate on biological activity at the human P2Y(2) receptor. Key steps in the synthesis of a series of 5-substituted uridine 5'-methylenephosphonates were the reaction of a suitably protected uridine 5'-aldehyde with [(diethoxyphosphinyl)methylidene]triphenylphosphorane, C-5 bromination and a Suzuki-Miyaura coupling. These analogues behaved as selective agonists at the P2Y(2) receptor, with three analogues exhibiting potencies in the submicromolar range. Although maximal activities observed with the phosphonate analogues were much less than observed with UTP, high concentrations of the phosphonates had no effect on the stimulatory effect of UTP. These results suggest that these phosphonates bind to an allosteric site of the P2Y(2) receptor.
Bioorganic & medicinal chemistry 02/2012; 20(7):2304-15. · 2.82 Impact Factor
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ABSTRACT: We present the use of 1-mm room-temperature probe technology to perform intermolecular interaction studies using chemical shift perturbation methods and saturation transfer difference (STD) spectroscopy using small sample volumes. The use of a small sample volume (5-10 µl) allows for an alternative titration protocol where individual samples are prepared for each titration point, rather than the usual protocol used for a 5-mm probe setup where the ligand is added consecutively to the solution containing the protein or host of interest. This allows for considerable economy in the consumption and cost of the protein and ligand amounts required for interaction studies. For titration experiments, the use of the 1-mm setup consumes less than 10% of the ligand amount required using a 5-mm setup. This is especially significant when complex ligands that are only available in limited quantities, typically because they are obtained from natural sources or through elaborate synthesis efforts, need to be investigated. While the use of smaller volumes does increase the measuring time, we demonstrate that the use of commercial small volume probes allows the study of interactions that would otherwise be impossible to address by NMR.
Magnetic Resonance in Chemistry 01/2011; 49(1):9-15. · 1.44 Impact Factor
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ABSTRACT: An efficient protocol for the construction of C-6-(hetero)aryl-substituted uridine phosphonate analogues utilizing an aerobic, ligand-free Suzuki-Miyaura cross-coupling reaction of a 6-iodo-precursor in aqueous media has been established. The method presents a modular approach toward the target compounds as demonstrated by the synthesis of a small library comprising 14 novel nucleoside phosphonates.
Organic & Biomolecular Chemistry 11/2010; 8(22):5234-46. · 3.70 Impact Factor
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ABSTRACT: Proton nuclear magnetic resonance ((1)H-NMR)-based metabolomics enables the high-resolution and high-throughput assessment of a broad spectrum of metabolites in biofluids. Despite the straightforward character of the experimental methodology, the analysis of spectral profiles is rather complex, particularly due to the requirement of numerous data preprocessing steps. Here, we evaluate how several of the most common preprocessing procedures affect the subsequent univariate analyses of blood serum spectra, with a particular focus on how the standard methods perform compared to more advanced examples. Carr-Purcell-Meiboom-Gill 1D (1)H spectra were obtained for 240 serum samples from healthy subjects of the Asklepios study. We studied the impact of different preprocessing steps--integral (standard method) and probabilistic quotient normalization; no, equidistant (standard), and adaptive-intelligent binning; mean (standard) and maximum bin intensity data summation--on the resonance intensities of three different types of metabolites: triglycerides, glucose, and creatinine. The effects were evaluated by correlating the differently preprocessed NMR data with the independently measured metabolite concentrations. The analyses revealed that the standard methods performed inferiorly and that a combination of probabilistic quotient normalization after adaptive-intelligent binning and maximum intensity variable definition yielded the best overall results (triglycerides, R = 0.98; glucose, R = 0.76; creatinine, R = 0.70). Therefore, at least in the case of serum metabolomics, these or equivalent methods should be preferred above the standard preprocessing methods, particularly for univariate analyses. Additional optimization of the normalization procedure might further improve the analyses.
Analytical and Bioanalytical Chemistry 10/2010; 398(4):1781-90. · 3.78 Impact Factor
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ABSTRACT: Polysorbate surfactants (commercially available as Tween) are widely used in pharmaceutical, cosmetic and food products. They are generally considered as esters of ethoxylated sorbitan with fatty acids. Diffusion H1 NMR spectroscopy on a solution of polysorbate 20 in D2O revealed that only one diffusion coefficient was found for the fatty acyl part. Using the Stokes-Einstein equation, it became obvious that this diffusion behavior was caused by micelles. On the other hand, two significantly different diffusion coefficients were found for the methylene groups of ethylene oxide (EO). This indicates the presence of two distinct EO containing species in solution. Since the slowest diffusing EO species has the same diffusion coefficient as the fatty acyl part, it corresponds to the micellar (i.e. fatty acyl bound) ethoxylates. The diffusion coefficient of the fastest diffusing EO species was a factor of four larger than that of the slowly diffusing species and was attributed to water-soluble non-esterified ethoxylates. A solution of polysorbate 20 in the presence of NaOD was prepared to investigate if hydrolysis of the sorbitan ester could be the reason for the occurence of these hydrophilic ethoxylates. It was found that alkaline hydrolysis does lead to an increasing fraction of non-esterified ethoxylates, but is not the cause of its presence in untreated polysorbate samples since these species were also found in solutions of polyethylene glycol oleyl ether (commercially available as Brij), which are not susceptible to hydrolysis. Fractionation of the EO species present in polysorbate 20 into an amphiphilic and a hydrophilic fraction was only partly obtained by activated carbon adsorption. On the other hand, sequential extraction of aqueous polysorbate solutions by ethyl acetate and chloroform enabled a nearly complete fractionation. H1 NMR spectroscopy proved to be very useful since it allows in situ determination of the global composition of a surfactant sample, as well as quantification of both the amphiphilic and hydrophilic ethoxylate fractions via diffusion measurements.
Journal of pharmaceutical and biomedical analysis 02/2010; 51(3):583-9. · 2.45 Impact Factor
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ABSTRACT: The influence of the physicochemical properties of the vesicular bilayer on the sorption of poorly water soluble compounds was investigated with pulsed field gradient 1H nuclear magnetic resonance (PFG-NMR) for the case of phosphatidylcholine and dioctadecyldimethylammonium bromide (DODAB), using 4-ethylbenzyl alcohol as a model compound. Hereby, the effect of bilayer thickness at a constant physicochemical state was studied using a range of phosphatidylcholines of varying chain lengths, whereas DODAB was preferred to check the influence of the bilayer physicochemical state since this cationic lipid is characterized by three different states within the studied temperature range. When the phospholipid alkyl chain length was changed, no differences were observed in the sorption which was linked to the surface-mediated sorption. On the other hand, when the chemical composition was preserved but the temperature and thus the physical state of the bilayer were changed, the sorption in dioctadecyldimethylammonium bromide (DODAB) vesicles changed dramatically. From those experiments, a strong relationship between the ordering of the surfactant molecules and the sorption can be assumed.
Langmuir 10/2009; 25(19):11322-7. · 4.19 Impact Factor
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ABSTRACT: Pseudodesmin A is a cyclic lipodepsipeptide (CLP) of the viscosin group with a moderate in vitro biological activity. For several CLPs, including members of this group, this activity has been related to the ability to form ion pores in cellular membranes. As their size does not allow individual CLPs to span the membrane bilayer, individual monomers must somehow assemble into a larger structure. NMR spectroscopy has been used to demonstrate that in chloroform and other apolar organic solvents, pseudodesmin A monomers assemble into a supramolecular structure. These self-assembled structures can become sufficiently large to span the membrane bilayer as demonstrated with translational diffusion NMR spectroscopic measurements. With the aim to obtain more insight into the structural nature of this assembly, the solution conformation of pseudodesmin A was first determined by using ROESY (rOe) restraints measured in acetonitrile, in which no self-association occurs. The structure, which is found to be mostly similar to the previously described crystal structure, is shown to be retained within the supramolecular complex. Intermolecular rOe contacts obtained in chloroform together with chemical shift perturbation data provides structural insight into the organization of the self-associated complex. Based upon this analysis, a model for the organization of pseudodesmin A monomers in the supramolecular assembly is proposed, which is in agreement with the formation of bilayer spanning hydrophilic pores and provides the basis for a structure-function relationship for this type of CLPs. Finally, it is demonstrated that the differences previously reported between the crystal and solution conformation of the white line inducing principle (WLIP), a close analogue of pseudodesmin A, are the result of the use of dimethyl sulfoxide as solvent, whose strong hydrogen-bonding capacity induces conformational exchange.
Chemistry 10/2009; 15(46):12653-62. · 5.93 Impact Factor
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ABSTRACT: The solubilization of the poorly water soluble anti-inflammatory drug flurbiprofen in non-ionic Tween 20 surfactant micellar solutions was studied by both (19)F and (1)H NMR spectroscopy in an acidic environment. These non-destructive techniques allowed us to investigate the effect of temperature cycling in situ. Using (19)F NMR, an increased solubilisation capacity was observed as the temperature increased. This effect became more pronounced above the cloud point, which was reduced by more than 30 degrees C in the presence of an excess of flurbiprofen. Upon clouding, peak splitting was observed in the (19)F spectrum, which indicates that two pools of solubilised flurbiprofen exist that are in slow exchange on the NMR frequency timescale. The clouding and solubilization processes were found to be reversible, albeit with slow kinetics. Based on chemical shift differences of both Tween 20 and flurbiprofen, as well as NOESY experiments, the flurbiprofen was found to be accumulated within the palisade layer of the Tween 20 micelles.
Physical Chemistry Chemical Physics 08/2009; 11(26):5462-8. · 3.57 Impact Factor
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ABSTRACT: Two new cyclic lipodepsipeptides named pseudodesmin A and B have been isolated from Pseudomonas bacteria collected from the mucus layer in the skin of the black belly salamander. Both compounds show moderate antibacterial activity against Gram positive bacteria, including MRSA. Complete 1 H, 13 C and 15 N NMR assignment of both compounds afforded their covalent structure and served to guide the analysis of LC-MS and X-ray diffraction data from which the final stereochemistry could be established. Both molecules can be categorized as new members of the viscosin group of cyclic lipodepsipeptides.
be (J. C. Martins) Tetrahedron. 01/2009; 65(21):4173-4181.
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ABSTRACT: As with every -omics technology, metabolomics requires new methodologies for data processing. Due to the large spectral size, a standard approach in NMR-based metabolomics implies the division of spectra into equally sized bins, thereby simplifying subsequent data analysis. Yet, disadvantages are the loss of information and the occurrence of artifacts caused by peak shifts. Here, a new binning algorithm, Adaptive Intelligent Binning (AI-Binning), which largely circumvents these problems, is presented. AI-Binning recursively identifies bin edges in existing bins, requires only minimal user input, and avoids the use of arbitrary parameters or reference spectra. The performance of AI-Binning is demonstrated using serum spectra from 40 hypertensive and 40 matched normotensive subjects from the Asklepios study. Hypertension is a major cardiovascular risk factor characterized by a complex biochemistry and, in most cases, an unknown origin. The binning algorithm resulted in an improved classification of hypertensive status compared with that of standard binning and facilitated the identification of relevant metabolites. Moreover, since the occurrence of noise variables is largely avoided, AI-Binned spectra can be unit-variance scaled. This enables the detection of relevant, low-intensity metabolites. These results demonstrate the power of AI-Binning and suggest the involvement of alpha-1 acid glycoproteins and choline biochemistry in hypertension.
Analytical Chemistry 06/2008; 80(10):3783-90. · 5.86 Impact Factor
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ABSTRACT: The sorption behavior of three benzyl alcohol derivatives with different hydrophobicities into sonicated and extruded DODAC dispersions has been studied using NMR spectroscopy and NMR diffusometry. We show that there is an increased sorption into a sonicated dispersion below the phase-transition temperature (T(m)) as compared to an extruded dispersion. This may be explained by the incomplete lipid chain freezing of charged lipids as a result of the sonication process. Around T(m), a sorption maximum is found that is attributed to the high bilayer disorder under this condition. In addition, a sorption increase and a fluidizing effect at increasing benzyl alcohol derivative concentrations are observed that provide additional evidence for the relevance of the bilayer fluidity on the sorption of hydrophobic components.
Langmuir 05/2008; 24(7):3082-9. · 4.19 Impact Factor
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Chemical Science. in-press:1-9.
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ABSTRACT: Interactions between phenol and cationic cetyltrimethylammonium bromide (CTAB) micelles have been investigated by means of nuclear magnetic resonance spectroscopy. The combined use of 1H and NOESY techniques revealed that phenol has different preferred locations of interaction depending on the pH. At neutral pH (6.70) conditions, phenol molecules are preferentially located in the outer micelle region, at the micelle–water interface, while at more basic pH (9.94), the deprotonated phenol molecules (C6H5O−) are immersed into the palisade layer of the micelle. In addition, quantitative estimates of the solubilized fraction of phenol were obtained by using PFG-NMR. The results indicate that the phenol–CTAB interactions, although already present in neutral pH conditions, are largely favored in basic conditions as a consequence of the strong electrostatic interaction between the negatively charged phenolate ions and the positive charge of the cationic surfactant head group.
Colloids and Surfaces A Physicochemical and Engineering Aspects 370:42-48. · 2.24 Impact Factor
