Johan Svenson

Can we rationally design molecularly imprinted polymers

The nearly exponential growth in the molecular imprinting literature has to a large extent been fuelled by an increasing awareness of the potential of molecular imprinting based technologies. Despite the acceptance of the technique by cognate disciplines and the demonstration of its usefulness in a number of enabling technologies, relatively little is known about the molecular level events underlying the imprinting process and subsequent recognition events. What rules govern imprint formation? Can we use such rules to rationally design molecularly imprinted polymers?

Synoxazolidinones A and B: novel bioactive alkaloids from the ascidian Synoicum pulmonaria.

Bioassay-guided fractionation of the sub-Arctic ascidian Synoicum pulmonaria collected off the Norwegian coast led to the isolation of a novel family of brominated guanidinium oxazolidinones named synoxazolidinones A and B (1 and 2). The backbone of the compounds contains a 4-oxazolidinone ring rarely seen in natural products. The structure of the compounds was determined by spectroscopic methods. The synoxazolidinones exhibited antibacterial and antifungal activities.

Determining the Absolute Configuration of Two Marine Compounds Using Vibrational Chiroptical Spectroscopy

Chiroptical techniques are increasingly employed for assigning the absolute configuration of chiral molecules through comparison of experimental spectra with theoretical predictions. For assignment of natural products, electronic chiroptical spectroscopies such as electronic circular dichroism (ECD) are routinely applied. However, the sensitivity of electronic spectral parameters to experimental conditions and the theoretical methods employed can lead to incorrect assignments. Vibrational chiroptical methods (vibrational circular dichroism, VCD, and Raman optical activity, ROA) provide more reliable assignments, although they, in particular ROA, have been little explored for assignments of natural products. In this study, the ECD, VCD, and ROA chiroptical spectroscopies are evaluated for the assignment of the absolute configuration of a highly flexible natural compound with two stereocenters and an asymmetrically substituted double bond, the marine antibiotic Synoxazolidinone A (SynOxA), recently isolated from the sub-Arctic ascidian Synoicum pulmonaria. Conformationally averaged nuclear magnetic resonance (NMR), ECD, Raman, ROA, infrared (IR) and VCD spectral parameters are computed for the eight possible stereoisomers of SynOxA and compared to experimental results. In contrast to previously reported results, the stereochemical assignment of SynOxA based on ECD spectral bands is found to be unreliable. On the other hand, ROA spectra allow for a reliable determination of the configuration at the double bond and the ring stereocenter. However, ROA is not able to resolve the chlorine-substituted stereogenic center on the guanidinium side chain of SynOxA. Application of the third chiroptical method, VCD, indicates unique spectral features for all eight SynOxA isomers in the theoretical spectra. Although the experimental VCD is weak and restricted by the limited amount of sample, it allows for a tentative assignment of the elusive chlorine-substituted stereocenter. VCD chiroptical analysis of a SynOxA derivative with three stereocenters, SynOxC, results in the same absolute configuration as for SynOxA. Despite the experimental challenges, the results convincingly prove that the assignment of absolute configuration based on vibrational chiroptical methods is more reliable than for ECD.

The structure of the fire fighting foam surfactant Forafac®1157 and its biological and photolytic transformation products.

For several decades, perfluorooctane sulfonate (PFOS) has widely been used as a fluorinated surfactant in aqueous film forming foams used as hydrocarbon fuel fire extinguishers. Due to concerns regarding its environmental persistence and toxicological effects, PFOS has recently been replaced by novel fluorinated surfactants such as Forafac®1157, developed by the DuPont company. The major component of Forafac®1157 is a 6:2 fluorotelomer sulfonamide alkylbetaine (6:2 FTAB), and a link between the trade name and the exact chemical structure is presented here to the scientific community for the first time. In the present work, the structure of the 6:2 FTAB was elucidated by (1)H, (13)C and (19)F nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry. Moreover, its major metabolites from blue mussel (Mytilus edulis) and turbot (Scophthalmus maximus) and its photolytic transformation products were identified. Contrary to what has earlier been observed for PFOS, the 6:2 FTAB was extensively metabolized by blue mussel and turbot exposed to Forafac®1157. The major metabolite was a deacetylated betaine species, from which mono- and di-demethylated metabolites also were formed. Another abundant metabolite was the 6:2 fluorotelomer sulfonamide. In another experiment, Forafac®1157 was subjected to UV-light induced photolysis. The experimental conditions aimed to simulate Arctic conditions and the deacetylated species was again the primary transformation product of 6:2 FTAB. A 6:2 fluorotelomer sulfonamide was also formed along with a non-identified transformation product. The environmental presence of most of the metabolites and transformation products was qualitatively demonstrated by analysis of soil samples taken in close proximity to an airport fire training facility.

A Combined Atomic Force Microscopy and Computational Approach for the Structural Elucidation of Breitfussin A and B: Highly Modified Halogenated Dipeptides from Thuiaria breitfussi

The use of atomic-force microscopy (AFM) with atomic resolution shows great potential for the structural characterization of planar, proton-poor compounds, as these compounds are prone to structural corrections. [1,2] Currently, AFM has limited ability to identify element type and consequently functional groups. Additional computational techniques, such as computer-aided structure elucidation (CASE) and the calculation of 13 C NMR shifts using electronic structure calculations (DFT) may assist in this respect. Herein we show the combined use of spectroscopic methods, AFM, CASE, and DFT to solve the structures of breitfussins A and B, which could not be solved using either method alone. The subject of this study was the Arctic hydrozoan Thuiaria breitfussi (Family Sertulariidae). The few publications on the chemistry of this family show the presence of sterols, [3] polyhalogenated monoterpenes, [4] and anthracenone derivatives. [5] Arctic marine environments support highly diverse and dense populations of marine invertebrates. [6,7] A

Spectroscopic studies of the molecular imprinting self-assembly process

A method for the rapid estimation of the extent of complex formation in molecular imprinting pre‐polymerization mixtures is described. By the use of a UV spectroscopy titration procedure, apparent binding constants for such self‐assembly processes have been obtained. This method was used for comparison of the interactions between a dipeptide template (N‐acetyl‐L‐phenylalaninyl‐L‐tryptophanyl methyl ester) and the functional monomer methacrylic acid, and the monomer analogues acetic acid and trifluoroacetic acid. The importance of template–monomer association during the molecular imprinting pre‐polymerization phase is discussed with respect to the systems studied. Copyright

Synthetic cationic antimicrobial peptides bind with their hydrophobic parts to drug site II of human serum albumin.

BackgroundMany biologically active compounds bind to plasma transport proteins, and this binding can be either advantageous or disadvantageous from a drug design perspective. Human serum albumin (HSA) is one of the most important transport proteins in the cardiovascular system due to its great binding capacity and high physiological concentration. HSA has a preference for accommodating neutral lipophilic and acidic drug-like ligands, but is also surprisingly able to bind positively charged peptides. Understanding of how short cationic antimicrobial peptides interact with human serum albumin is of importance for developing such compounds into the clinics.ResultsThe binding of a selection of short synthetic cationic antimicrobial peptides (CAPs) to human albumin with binding affinities in the μM range is described. Competitive isothermal titration calorimetry (ITC) and NMR WaterLOGSY experiments mapped the binding site of the CAPs to the well-known drug site II within subdomain IIIA of HSA. Thermodynamic and structural analysis revealed that the binding is exclusively driven by interactions with the hydrophobic moieties of the peptides, and is independent of the cationic residues that are vital for antimicrobial activity. Both of the hydrophobic moieties comprising the peptides were detected to interact with drug site II by NMR saturation transfer difference (STD) group epitope mapping (GEM) and INPHARMA experiments. Molecular models of the complexes between the peptides and albumin were constructed using docking experiments, and support the binding hypothesis and confirm the overall binding affinities of the CAPs.ConclusionsThe biophysical and structural characterizations of albumin-peptide complexes reported here provide detailed insight into how albumin can bind short cationic peptides. The hydrophobic elements of the peptides studied here are responsible for the main interaction with HSA. We suggest that albumin binding should be taken into careful consideration in antimicrobial peptide studies, as the systemic distribution can be significantly affected by HSA interactions.

Antifouling compounds from the sub-arctic ascidian Synoicum pulmonaria: synoxazolidinones A and C, pulmonarins A and B, and synthetic analogues.

The current study describes the antifouling properties of four members belonging to the recently discovered synoxazolidinone and pulmonarin families, isolated from the sub-Arctic sessile ascidian Synoicum pulmonaria collected off the Norwegian coast. Four simplified synthetic analogues were also prepared and included in the study. Several of the studied compounds displayed MIC values in the micro-nanomolar range against 16 relevant marine species involved in both the micro- and macrofouling process. Settlement studies on Balanus improvisus cyprids indicated a deterrent effect and a low toxicity for selected compounds. The two synoxazolidinones displayed broad activity and are shown to be among the most active natural antifouling bromotyrosine derivatives described. Synoxazolidinone C displayed selected antifouling properties comparable to the commercial antifouling product Sea-Nine-211. The pulmonarins prevented the growth of several bacterial strains at nanomolar concentrations but displayed a lower activity toward microalgae and no effect on barnacles. The linear and cyclic synthetic peptidic mimics also displayed potent antifouling activities mainly directed against bacterial adhesion and growth.

Metabolic Fate of Lactoferricin Based Antimicrobial Peptides: Effect of Truncation and Incorporation of Amino Acid Analogs on the in vitro Metabolic Stability

A series of promising truncated antibacterial tripeptides derived from lactoferricin has been prepared, and their in vitro metabolic stability in the main metabolic compartments, plasma, liver, kidney, stomach, duodenum, and brain, has been investigated for the first time. The potential stabilizing effect of truncation, C-terminal capping, and introduction of the bulky synthetic amino acid biphenylalanine is also investigated. The drug-like peptides displayed large differences in half-lives in the different matrixes ranging from 4.2 min in stomach and duodenum to 355.9 min in liver. Kinetic analysis of the metabolites revealed that several different degrading enzymes simultaneously target the different peptide bonds and that the outcome of the tested strategies to increase the stability is clearly enzyme-specific. Some of the metabolic enzymes even prefer the synthetic modifications incorporated over the natural counterparts. Collectively, it is shown that the necessary antibacterial pharmacophore generates compounds that are not only potent antibacterial peptides, but excellent substrates for the main degrading enzymes. All the amide bonds are thus rapidly targeted by different enzymes despite the short peptidic sequences of the tested compounds. Hence, our results illustrate that several structural changes are needed before these compounds can be considered for oral administration. Strategies to overcome such metabolic challenges are discussed.

Altered activity and physicochemical properties of short cationic antimicrobial peptides by incorporation of arginine analogues.

The incorporation of nongenetically encoded amino acids is a well established strategy to alter the behavior of several types of promising cationic antimicrobial peptides. Generally, these elements have been improved mimics of the hydrophobic amino acids yielding peptides with increased stability and potency. In this initial study, the effect of systematic replacement of Arg in a well-defined moderately antimicrobial tripeptide library is described. It is shown that the arginine analogues need to display a strong basicity to produce active peptides. It is further revealed that the hydrophobic units needed for activity in these peptides can be effectively incorporated in the direct vicinity of the cationic charge to produce compounds with improved antibacterial properties. A well-defined facial amphiphilic structure, which remains intact upon introduction of hydrophobic elements in the cationic side chains, is seen for the majority of the tested peptides. Microcalorimetric studies revealed a peptide binding to large anionic unilamellar vesicles (LUVs) mimicking the Gram-positive bacterial membrane as well as a potentially competitive binding to human serum albumin in the low- to mid-micromolar range. No considerable alterations in binding to either albumin or the LUVs were seen for the analogue containing peptides. A neutral LUV mimicking the eukaryotic cell membrane showed no significant binding to any of the peptides. The oral absorption of this class of short lactoferricin based peptides was investigated for the first time and revealed that incorporation of weaker bases than Arg produced peptides with much improved permeability in a recently developed permeation model, the phospholipid vesicle based barrier assay. Collectively, the results presented here show that there is ample room to toggle the activity and physical properties of short cationic antimicrobial peptides by incorporation of arginine analogues.