Surface modification of polymeric micelles by strain-promoted alkyne-azide cycloadditions

Organomicelles modified by surface dibenzylcyclooctyne moieties can conveniently be functionalized by strain-promoted alkyne-azide cycloadditions. The ligation approach is highly efficient, does not require toxic reagents and is compatible with a wide variety of functional modules. Interactions of proteins with surface ligands of the micelles have been studied by AFM, which revealed that it leads to disassembly of the particles thereby providing a mechanism for triggered drug release.     

Transition states of strain-promoted metal-free click chemistry: 1,3-dipolar cycloadditions of phenyl azide and cyclooctynes

Density functional theory (B3LYP) calculations on the transition states for the Huisgen 1,3-dipolar cycloadditions of phenyl azide with acetylene, cyclooctyne, and difluorocyclooctyne are reported. The low activation energy of the cyclooctyne "strain-promoted" cycloaddition (DeltaE = 8.0) compared to the strain-free acetylene cycloaddition (DeltaE = 16.2) is due to decreased distortion energy (DeltaEd) of cyclooctyne (DeltaDeltaEd = 4.6) and phenyl azide (DeltaDeltaEd = 4.5) to achieve that cycloaddition transition state. Electronegative fluorine substituents on cyclooctyne further increase the rate of cycloaddition by increasing interaction energies.     

Cyclizations and cycloadditions of acetylenic sulfones on solid supports

Acetylenic sulfones attached to solid supports by means of ester linkers were employed in a variety of cyclization and cycloaddition reactions, followed by cleavage of the products from the resin by ester hydrolysis or reductive desulfonylation.     

Peptidotriazoles on solid phase: [1,2,3]-triazoles by regiospecific copper(i)-catalyzed 1,3-dipolar cycloadditions of terminal alkynes to azides

The cycloaddition of azides to alkynes is one of the most important synthetic routes to 1H-[1,2,3]-triazoles. Here a novel regiospecific copper(I)-catalyzed 1,3-dipolar cycloaddition of terminal alkynes to azides on solid-phase is reported. Primary, secondary, and tertiary alkyl azides, aryl azides, and an azido sugar were used successfully in the copper(I)-catalyzed cycloaddition producing diversely 1,4-substituted [1,2,3]-triazoles in peptide backbones or side chains. The reaction conditions were fully compatible with solid-phase peptide synthesis on polar supports. The copper(I) catalysis is mild and efficient (>95% conversion and purity in most cases) and furthermore, the X-ray structure of 2-azido-2-methylpropanoic acid has been solved, to yield structural information on the 1,3-dipoles entering the reaction. Novel Fmoc-protected amino azides derived from Fmoc-amino alcohols were prepared by the Mitsunobu reaction.     

1,3-Dipolar cycloadditions of acetylenic sulfones in solution and on solid supports

Several representative acetylenic sulfones were immobilized on a polymer support derived from Merrifield resin by means of ester linkers that were used to couple free carboxylic acid groups on the solid support with benzylic hydroxyl functions on the arylsulfonyl moieties of the acetylenes. Several examples of reversed ester linkers, using Merrifield resin directly, were also successfully prepared. The 1,3-dipolar cycloadditions of the solid-supported acetylenic sulfones were investigated with a series of 1,3-dipoles, including benzyl azide, ethyl diazoacetate, diazomethane, as well as representative nitrile oxides, nitrile imines, nitrile ylides, nitrones, azomethine imines, azomethine ylides, munchnones, and sydnones. In general, analogous cycloadditions were also performed with acetylenic sulfones in solution phase for comparison. The cycloadditions typically afforded good to excellent yields of the desired products in both solution and solid phase, although the latter reactions sometimes required more vigorous conditions. Except in the case of benzyl azide and diazo compounds, where mixtures of regioisomers were obtained, the other 1,3-dipoles reacted with high regioselectivity and afforded essentially unique regioisomers. Cleavage of the products from the resin was smoothly effected by alkaline hydrolysis, while several attempts at reductive desulfonylation with sodium amalgam or samarium diiodide-HMPA resulted in N-O or C-O scission, in addition to cleavage from the polymer. The method provides access to a number of important classes of heterocycles, including variously substituted and functionalized triazoles, pyrazoles, 1,2-oxazoles, pyrroles, as well as their dihydro and bicyclic analogues. The success of the cycloadditions on polymer supports paves the way to future investigations of sequential transformations leading to libraries of useful heterocycles.     

Fast folding of RNA pseudoknots initiated by laser temperature-jump

RNA pseudoknots are examples of minimal structural motifs in RNA with tertiary interactions that stabilize the structures of many ribozymes. They also play an essential role in a variety of biological functions that are modulated by their structure, stability, and dynamics. Therefore, understanding the global principles that determine the thermodynamics and folding pathways of RNA pseudoknots is an important problem in biology, both for elucidating the folding mechanisms of larger ribozymes as well as addressing issues of possible kinetic control of the biological functions of pseudoknots. We report on the folding/unfolding kinetics of a hairpin-type pseudoknot obtained with microsecond time-resolution in response to a laser temperature-jump perturbation. The kinetics are monitored using UV absorbance as well as fluorescence of extrinsically attached labels as spectroscopic probes of the transiently populated RNA conformations. We measure folding times of 1-6 ms at 37 °C, which are at least 100-fold faster than previous observations of very slow folding pseudoknots that were trapped in misfolded conformations. The measured relaxation times are remarkably similar to predictions of a computational study by Thirumalai and co-workers (Cho, S. S.; Pincus, D.L.; Thirumalai, D. Proc. Natl. Acad. Sci. U. S. A. 2009, 106, 17349-17354). Thus, these studies provide the first observation of a fast-folding pseudoknot and present a benchmark against which computational models can be refined.     

Fast solid phase extraction of polychlorobiphenyls and chlorinated pesticide residues from mussels using sep-pak cartridges

Summary A rapid method for the determination of chlorinated pesticides and polychlorinated biphenyls in mussels (Mytilus sp.) is reported. The mussel sample is homogenized and extracted with acetonitrile. The organic solution is concentrated and successively diluted with distilled water solution (12 g L⁻¹ NaCl). The organic compounds from water solution are adsorbed onto a NH₂ Sep-Pak cartridge. The clean-up step, in which the polychlorobiphenyls and chiorinated pesticides are separated in different eluates, is achieved by passing 25 mL of a 40% methanol aqueous solution through the NH₂ Sep-Pak and the C₁₈ Sep-Pak cartridges connected in series. The polychloroblphenyls are desorbed from the NH₂ Sep-Pak cartridge whilst the chlorinated peslicides are recovered from the C₁₈ Sep-Pak cartridge. In the separation of polychlorobiphenyls from the chlorinated pesticides tested in this work, only aldrin, hepatachlor and 4,4′-DDD are partially adsorbed with the polychlorobiphenyls onto the NH₂ Sep-Pak cartridge. The average recovery is ≥95.0% with a relative standard deviation ≤5.0%. The limits of detection for different pesticides and polychlorobiphenyl congeners are 0.01 and 0.008 μg Kg⁻¹. The final determination is carried out by capillary gas chromatography with ECD.     

Enantioselective synthesis on the solid phase

Organic transformations on substrates which are immobilized on an insoluble, polymeric carrier have found broad application in compound collection synthesis. In contrast to other synthetic methodologies in solid-phase organic synthesis, reactions that afford non-racemic products are strikingly under-represented. Not only does the introduction of stereoinformation on immobilized, achiral starting materials provide enantioenriched products which can be of value for biological testing, but it also opens up new perspectives for accessible structures. This feature article gives an overview of successful enantioselective transformations on a solid support. Critical differences in the corresponding solution-phase protocols are highlighted, and applications to the generation of compound collections are particularly mentioned.     

Synthesis of cyclic peptides by solid phase methodology

Four carba analogues of oxytocin and a cyclic analogue of melanocyte stimulating hormone were synthesized using solid phase methodology. Purified compounds were shown to be highly biologically active.     

Fast determination of hydroxylated polychlorinated biphenyls in human plasma by online solid phase extraction coupled to liquid chromatography-tandem mass spectrometry

Hydroxylated polychlorinated biphenyls (OH-PCBs) have been shown to be strongly retained in human blood causing endocrine-related toxicity, particularly on the thyroid system. Traditionally, analytical methods for the determination of OH-PCBs require labor-intensive and long-time consuming sample preparation with several extraction, evaporation and cleanup procedures steps and, in some cases, derivatization prior to the analysis by gas or liquid chromatography-mass spectrometry (GC–MS or LC-MS). The present study developed and validated a novel, sensitive and high throughput online solid phase extraction (SPE) method coupled to LC-tandem mass spectrometry (MS/MS) for the separation and quantitation of relevant congeners of OH-PCBs in human plasma. The developed method presented limits of quantification (LOQ) ranging from 0.02 to 0.5 ng mL⁻¹ and extraction recoveries from 71 to 134% for all congeners, requiring small amount of sample (only 100 μL) and minimal sample preparation. In order to evaluate the applicability of the method, preliminary tests (N = 93) were conducted in plasma from individuals occupationally exposed to very high levels of PCBs in a German cohort. Penta-through hepta-chlorinated OH-PCBs were the predominant congeners in human plasma with concentrations up to 44.5 ng mL⁻¹, while lower chlorinated OH-PCBs were occasionally detected. In addition, a new PCB 28 metabolite has been synthesized and identified for the first time in human plasma and associations between OH-PCBs and their parent compounds in the studied cohort were also assessed.     

Parallel synthesis of H-pin polyamides by alkene metathesis on solid phase

A small library of H-pin polyamides with variable aliphatic bridge lengths (CH(2))(n)(), where n = 4-8, connecting a central Py/Py pair was prepared via parallel synthesis with Ru-catalyzed alkene metathesis on solid phase as a complexity-generating cross-linking reaction. DNA binding affinities and sequence specificities were analyzed for each member of the library to determine the optimum linker length. An H-pin polyamide with a six-methylene bridge was found to have the highest affinity to its match site with high selectivity over a 1-bp mismatch site. The relationship between the number of methylenes in the linker (CH(2))(n)() and affinity is n = 6 > 4 > 7 > 5 > 8. These results indicate that 6 followed by 4 methylene-bridged polyamides represent the optimum spacer length for the H-pin motif in the DNA minor groove. Importantly, the H-pin is competitive with hairpin polyamides with respect to affinity and specificity. The metathesis-based convergent synthetic route to H-pin polyamides expands the scope of readily available DNA recognition motifs for small molecule-based gene regulation studies.     

Site-specific conjugation of ScFvs antibodies to nanoparticles by bioorthogonal strain-promoted alkyne-nitrone cycloaddition

Particularly suitable: An N-terminal serine mutant of anti-HER2 scFv antibody was conjugated to polymer-coated magnetofluorescent nanoparticles by strain-promoted alkyne-nitrone cycloaddition. The resulting nanoparticles (see scheme) proved effective in targeting and labeling HER2-positive breast cancer cells.     

Fast detection of methyl tert-butyl ether from water using solid phase microextraction and ion mobility spectrometry

Methyl tert-butyl ether (MTBE) is commonly used as chemical additive to increase oxygen content and octane rating of reformulated gasoline. Despite its impact on enhancing cleaner combustion of gasoline, MTBE poses a threat to surface and ground water when gasoline is released into the environment. Methods for onsite analysis of MTBE in water samples are also needed. A less common technique for MTBE detection from water is ion mobility spectrometry (IMS). We describe a method for fast sampling and screening of MTBE from water by solid phase microextraction (SPME) and IMS. MTBE is adsorbed from the head space of a sample to the coating of SPME fiber. The interface containing a heated sample chamber, which couples SPME and IMS, was constructed and the SPME fiber was introduced into the sample chamber for thermal desorption and IMS detection of MTBE vapors. The demonstrated SPME-IMS method proved to be a straightforward method for the detection of trace quantities of MTBE from waters including surface and ground water. We determined the relative standard deviation of 8.3% and detection limit of 5 mg L⁻¹ for MTBE. Because of short sampling, desorption, and detection times, the described configuration of combined SPME and IMS is a feasible method for the detection of hazardous substances from environmental matrices.     

Multi-residue-analysis of pesticides by HPLC after solid phase extraction

Summary HPLC is a good method for analyzing a broad variety of different pesticides in drinking and surface water. Due to a high enrichment factor in solid phase extraction and the sensitive detection in the UV-range many pesticides can be determined even at a concentration lower than 0.05 g/l. By the use of a diode-array-detector a good security is reached for the identification of substances.     

Iron-assisted nucleophilic aromatic substitution on solid phase

Iron-assisted S(N)Ar reactions were performed for the first time on solid phase, and a library of 36 unsymmetrically substituted phenylpiperazines and phenyl-1,4-diazepanes was synthesized with this novel strategy. The scope of iron-assisted S(N)Ar reactions on solid phase was investigated, and reactions of representative nucleophiles from groups VI (O, S, and Se) and V (N and P) of the periodic table were examined. Decomplexation of resin-bound iron complexes was achieved with 1,10-phenanthroline under irradiation, thereby overcoming the notorious disadvantages of decomplexation observed in solution-phase chemistry.     

Process structuring of polymers by solid phase orientation processing

Solid phase orientation of polymers is one of the most successful routes to enhancement of polymer properties.It unlocks the potential of molecular orientation for the achievement of a range of enhanced physical properties.We provide here an overview of techniques developed in our laboratories for structuring polymers by solid phase orientation processing routes,with a particular focus on die drawing,which have allowed control of significant enhancements of a single property or combinations of properties,including Young’s modulus,strength,and density.These have led to notable commercial exploitations,and examples of load bearing low density materials and shape memory materials are discussed.