Fluorous tag method for the simultaneous synthesis of different kinds of glycolipids

A mixture of saccharide primers with partially fluorinated tails, 2-(perfluorooctyl)ethyl 4′-O-(β-d-galactopyranosyl)-β-d-glucopyranoside (Lac H2F8) and 6-(perfluorohexyl)hexyl 2′-acetamido-2′-deoxy-β-d-glucopyranoside (GN H6H6), were introduced to animal cells. The oligosaccharide of Lac H2F8 was elongated by cellular enzymes and gave a GM3-type oligosaccharide. On the other hand, GN H6F6 was galactosylated to afford a lactosamine derivative that was further sialylated. This research confirmed that simultaneous glycosylation processes took place for Lac H2F8 and GN H6F6 primers and that the presence of one did not prevent the glycosylation of the other from proceeding. Each primer was recognized independently and elongated sequentially by cellular enzymes. Significantly, the synthesis of glycolipids from a mixture of these artificial scaffolds did not prevent the synthesis of glycolipids from the natural precursor. The glycosyl transferases recognized both precursors resulting to simultaneous synthesis of glycolipids.     

α‐ and β‐Lipomycin: Total Syntheses by Sequential Stille Couplings and Assignment of the Absolute Configuration of All Stereogenic Centers

40 years ago spectroscopy, derivatization, and degradation revealed the structures of α‐lipomycin and its aglycon β‐lipomycin except for the configurations of their side‐chain stereocenters. We synthesized all relevant β‐lipomycin candidates: the (12R,13S) isomer has the same specific rotational value as the natural product. By the same criterion the (12R,13S)‐configured D ‐digitoxide is identical to α‐lipomycin. We double‐checked our assignments by degrading α‐ and β‐lipomycin to the diesters 33 and 34 and proving their 3D structures synthetically.     

Total Synthesis of Sialylated Glycans Related to Avian and Human Influenza Virus Infection

Human and avian influenza type A viruses bind sialylated pentasaccharides. Herein, the total synthesis of four of these glycans is reported. Efficient sialylations relied on two N‐Troc‐protected (Troc=2,2,2‐trichloroethoxycarbonyl) sialic acid building blocks. The first, a thiophenyl glycoside, readily produced the sialyl‐α(2‐6)galactose disaccharide. Combination of the second building block, a novel glycosyl phosphite, and a benzylidene‐protected galactoside produced the best results for the formation of the sialyl‐α(2‐3)galactose. Two common trisaccharides were assembled by the introduction of glucose, galactose, and glucosamine building blocks followed by selective deprotection. Two sets of pentasaccharides were obtained by the union of two sialylgalactose N‐phenyl trifluoroacetimidate building blocks with the two trisaccharides above. Global deprotection furnished the desired pentasaccharides. The products of these total syntheses are currently employed on the surface of carbohydrate microarrays to detect and type different strains of the influenza virus.     

A Novel Method for the Production of Glycosphingolipids

Neolacto‐series ganglioside sialylparagloboside (SPG) is a ganglioside species present in various human tissues, and used in many important studies. In this study, four ganglioside analogs, GM3, GD3, SPG, and NeuAc‐Gal‐GlcNAc‐Gal‐GlcNAc‐Gal‐Glc‐Cer, were synthesized by the saccharide‐primer method using MDCK cells and β‐lactoside primer with different aglycons. As compared to former methods for producing SPG, the primer method was rapid and convenient. Moreover, the yield of SPG was much higher than that obtained by former methods. The production of gangliosides with an azido group in the aglycon moiety was also achieved by using MDCK cells.     

Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE‐ESI‐MS

A glycosphingolipid analogue (12‐azidododecyl β‐lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE‐ESI‐MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12‐azidododecyl β‐lactoside (Lac‐C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac‐C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CE‐ESI‐MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.     

A total synthesis of the methyl glycoside of ganglioside GM(1)

The total synthesis of the methyl glycoside of GM(1) (1b) has been accomplished. The key step in the synthesis involves the sulfonamidoglycosidation reaction, which is used to create a beta-linkage leading to a GalNAc residue joined to the C4 hydroxyl group of a galactose unit of a C3 sialylated lactosyl moiety. The "proximal hydroxyl" directing effect, which has been postulated before, manifests in this context as well leading to the preponderant formation of the beta-glycoside. Together with asialo GM(1) and other substructures, the GM(1) methyl glycoside has been submitted for biological assays as potential ligands for bacterial and viral infection sites.     

Tumor suppression via diverting intracellular sialylation with multifunctional nanoparticles

Sialylation plays an important role in tumor-related physiological processes. Therefore, intervention of sialylation has great potential to explore new paths for tumor therapy. In view of the immune modulation of sialic acid (SA) on tumors, this work designs a multifunctional mesoporous silica nanoparticle (MFMSN) to divert intracellular sialylation for tumor suppression. The galactose groups covered on MFMSN act as sialylation substrates to bind intracellular SAs competitively, which inhibits the SA expression on the tumor cell surface. The diverted intracellular sialylation can be visualized on living cells and in vivo by specifically binding the sialylated galactose with a phenylboronic acid labeled ssDNA probe released from the pore of MFMSN to induce DNA strand displacement, which recovers the fluorescence of the dsDNA probe covered on MFMSN surface. The diverting of sialylation efficiently suppresses tumor growth in mice, demonstrating the great potential of the designed strategy for revealing SA-related biological processes and clinical cancer therapy.

Multifunctional nanoparticles are designed to divert intracellular sialylation, which can suppress tumor growth and be visualized.     

Microscopic Fourier Transform Infrared Characterization on Two Types of Spherulite with Polymorphic Crystals in Poly(heptamethylene terephthalate)

FTIR microspectrometry with in situ temperature variation and IR‐peak‐mapping capability, and POM characterization were used to study the crystal distribution in dual spherulites in poly(heptamethylene terephthalate). By tracing the crystalline IR bands of the α‐crystal and β‐crystal to get the crystal distribution, the techniques resolve that the ringed and ringless spherulites comprise α‐ and β‐crystals, respectively. In addition, temperature‐dependent IR analyses on the spots related to the two crystals also reveal the α‐ and β‐crystals melt at 98 and 104 °C, respectively. The ringed and ringless spherulites were proven to be correlated with the α‐ and β‐crystal forms, respectively.

     

Synthesis and Cytotoxic Activity of Macromolecular Prodrug of Cisplatin Using Poly(ethylene glycol) with Galactose Residues or Antennary Galactose Units

To provide a macromolecular prodrug with recognition ability for hepatoma cells, we synthesized new conjugates of cisplatin (CDDP) and poly(ethylene glycol) (PEG) with galactose residues or antennary galactose units (Gal4A, four branched galactose residues) at the chain terminus, Gal‐PEG‐DA/CDDP or Gal4A‐PEG‐DA/CDDP conjugates. An antennary (branched) structure of Gal4A was designed based on the fact that saccharide clusters with branched structures show highly effective binding with saccharide receptors, a phenomenon known as the ‘cluster effect’. The cytotoxic activity of the conjugates was investigated against HepG2 human hepatoma cells in vitro and compared with a control conjugate without galactose, MeO‐PEG‐DA/CDDP. Gal‐PEG‐DA/CDDP and Gal4A‐PEG‐DA/CDDP conjugates showed lower IC₅₀ values (3.1×10^–4 and 2.3×10^–4 M , respectively) than the MeO‐PEG‐DA/CDDP conjugate (10.5×10^–4 M ). The cytotoxic activities of these conjugates with galactose residues or antennary galactose units were inhibited as a result of the addition of galactose and strongly inhibited by the addition of Gal4A, however the inclusion of a methoxy group (the MeO‐PEG‐DA/CDDP conjugate) did not affect the activity. These results suggest that the Gal4A unit introduced to the conjugate has effective recognition ability against HepG2 human hepatoma cells.     

Monosaccharides as Versatile Units for Water‐Soluble Supramolecular Polymers

We introduce monosaccharides as versatile water‐soluble units to compatibilise supramolecular polymers based on the benzene‐1,3,5‐tricarboxamide (BTA) moiety with water. A library of monosaccharide‐based BTAs is evaluated, varying the length of the alkyl chain (hexyl, octyl, decyl and dodecyl) separating the BTA and saccharide units, as well as the saccharide units (α‐glucose, β‐glucose, α‐mannose and α‐galactose). In all cases, the monosaccharides impart excellent water compatibility. The length of the alkyl chain is the determining factor to obtain either long, one‐dimensional supramolecular polymers (dodecyl spacer), small aggregates (decyl spacer) or molecularly dissolved (octyl and hexyl) BTAs in water. For the BTAs comprising a dodecyl spacer, our results suggest that a cooperative self‐assembly process is operative and that the introduction of different monosaccharides does not significantly change the self‐ assembly behaviour. Finally, we investigate the potential of post‐assembly functionalisation of the formed supramolecular polymers by taking advantage of dynamic covalent bond formation between the monosaccharides and benzoxaboroles. We observe that the supramolecular polymers readily react with a fluorescent benzoxaborole derivative permitting imaging of these dynamic complexes by confocal fluorescence microscopy.     

Thermodynamic Phase Transition Through Crystal‐to‐Crystal Process of Photochromic 1,2‐Bis(5‐phenyl‐2‐propyl‐3‐thienyl)perfluorocyclopentene

A photochromic diarylethene, 1,2‐bis(5‐phenyl‐2‐propyl‐3‐thienyl)perfluorocyclopentene ( 1a ), was found to have two polymorphic crystal forms, α‐ and β‐crystals. From X‐ray crystallographic analysis, the space groups of α‐ and β‐crystals were determined to be P2₁/c and C2/c, respectively. The difference between two crystal forms is ascribed to the orientation of two of four molecules in the unit cell. The thermodynamic phase transition from α‐ to β‐forms occurred via a crystal‐to‐crystal process, as confirmed by differential scanning calorimetry measurements, optical microscopic observations in the reflection mode and under crossed Nicols, and powder X‐ray diffraction measurements. The movement of the molecules in the crystal was evaluated by analyzing the change of face indices before and after the phase transition.     

Determination of Danofloxacin and Marbofloxacin in Milk Samples by Micellar Liquid Chromatography with Fluorescence Detection

Abstract

A simple, rapid, and sensitive analytical method has been developed for the determination of two fluoroquinolones, danofloxacin and marbofloxacin, in bovine milk samples. Separation and quantification were performed by micellar liquid chromatography with fluorescence detection (MLC?FD), using sodium dodecyl sulfate (SDS) as a surfactant. The influence of the principal factors, namely, the micelle concentration, the amount of organic modifier, tail‐reducing agents, the pH, and the temperature were studied. The suitable condition was found to be 75 mM SDS?10 mM phosphate buffer–18 mM tetrabutylammonium bromide/3% (v/v) 1‐propanol at pH 3.0 for the separation of marbofloxacin, danofloxacin, and tosufloxacin (internal standard) in about 20 min. The linear concentration range of application was 1.8–30.0 ng · mL^?1 for danofloxacin and 16–120 ng · mL^?1 for marbofloxacin, and the relative standard deviation ranged between 4.9 and 2.7%. The limit of detection found for danofloxacin was 0.5 ng · mL^?1 and 5 ng · mL^?1 for marbofloxacin. These values were lower than the maximum residue limits (MRLs) established by the European Union for these compounds in bovine milk. It was applied to check the eventual existence of these compounds above these limits on commercial milk samples. The validation method was completed with spiked milk samples. Recovery levels obtained were 90.3–108.2%.     

The structure of the complex of cellulose I with ethylenediamine by X-ray crystallography and cross-polarization/magic angle spinning 13C nuclear magnetic resonance

X-ray crystallographic and cross-polarization/magic angle spinning ¹³C nuclear magnetic resonance techniques have been used to study an ethylenediamine (EDA)-cellulose I complex, a transient structure in the cellulose I to cellulose III_I conversion. The crystal structure (space group P2 ₁ ; a = 4.546 Å, b = 11.330 Å, c = 10.368 Å and γ = 94.017°) corresponds to a one-chain unit cell with one glucosyl residue in the asymmetric unit, a gt conformation for the hydroxymethyl group, and one EDA molecule per glucosyl residue. Unusually, there are no O–H···O hydrogen bonds between the cellulose chains; the chains are arranged in hydrophobic stacks, stabilized by hydrogen bonds to the amine groups of bridging EDA molecules. This new structure is an example of a complex in which the cellulose chains are isolated from each other, and provides a number of insights into the structural pathway followed during the conversion of cellulose I to cellulose III_I through EDA treatment.     

Bioethanol Production from Soybean Residue via Separate Hydrolysis and Fermentation

Bioethanol was produced using polysaccharide from soybean residue as biomass by separate hydrolysis and fermentation (SHF). This study focused on pretreatment, enzyme saccharification, and fermentation. Pretreatment to obtain monosaccharide was carried out with 20% (w/v) soybean residue slurry and 270 mmol/L H₂SO₄ at 121 °C for 60 min. More monosaccharide was obtained from enzymatic hydrolysis with a 16 U/mL mixture of commercial enzymes C-Tec 2 and Viscozyme L at 45 °C for 48 h. Ethanol fermentation with 20% (w/v) soybean residue hydrolysate was performed using wild-type and Saccharomyces cerevisiae KCCM 1129 adapted to high concentrations of galactose, using a flask and 5-L fermenter. When the wild type of S. cerevisiae was used, an ethanol production of 20.8 g/L with an ethanol yield of 0.31 g/g consumed glucose was obtained. Ethanol productions of 33.9 and 31.6 g/L with ethanol yield of 0.49 g/g consumed glucose and 0.47 g/g consumed glucose were obtained in a flask and a 5-L fermenter, respectively, using S. cerevisiae adapted to a high concentration of galactose. Therefore, adapted S. cerevisiae to galactose could enhance the overall ethanol fermentation yields compared to the wild-type one.     

Effect of cosolvent on solid phase transitions of α‐ to β‐form crystal of syndiotactic polystyrene occurred in supercritical Co2 containing solvent

The solid phase transition mechanism of α‐ to β‐form crystal upon specific treating with supercritical CO₂ + cosolvent on original pure α and mixed (α+β) form syndiotactic polystyrene (sPS) was investigated, using wide angle X‐ray diffraction and differential scanning calorimetry measurements as a function of temperature, pressure, and cosolvent content. As in the supercritical CO₂, sPS in supercritical CO₂ + cosolvent underwent solid phase transitions from α‐ to β‐form, and higher temperature or higher pressure favored this transformation. Due to the higher dipole moment of acetone, small amounts of acetone used as cosolvent with CO₂ made the transition of α‐ to β‐form occur at lower temperature and pressure than in supercritical CO₂, and made the α‐form crystal completely transform to β‐form in the original mixed (α+β) form, whereas ethanol did not. The original β‐form crystal in the original mixed (α+β) form sample acted as the nucleus of new β‐form crystal in the presence of cosolvent as it did in supercritical CO₂, when compared with the original pure α‐form sample. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1625–1636, 2007     

Stereoselective Synthesis of α‐ and β‐Glycofuranosyl Amides by Traceless Ligation of Glycofuranosyl Azides

A highly stereoselective synthesis of α‐ or β‐glycofuranosyl amides based on the traceless Staudinger ligation of glycofuranosyl azides of the galacto, ribo, and arabino series with 2‐diphenylphosphanyl‐phenyl esters has been developed. Both α‐ and β‐isomers can be obtained with excellent selectivity from a common, easily available precursor. The process does not depend on the anomeric configuration of the starting azide but appears to be controlled by the C2 configuration and by the protection/deprotection state of the substrates. A mechanistic interpretation of the results, supported by ³¹P NMR experiments, is offered and merged with our previous mechanistic analysis of pyranosyl azide ligation reactions.     

Selective Ablation of β‐Galactosidase‐Expressing Cells with a Rationally Designed Activatable Photosensitizer

We have developed an activatable photosensitizer capable of specifically inducing the death of β‐galactosidase‐expressing cells in response to photoirradiation. By using a selenium‐substituted rhodol scaffold bearing β‐galactoside as a targeting substituent, we designed and synthesized HMDESeR‐βGal, which has a non‐phototoxic spirocyclic structure owing to the presence of the galactoside moiety. However, β‐galactosidase efficiently converted HMDESeR‐βGal into phototoxic HMDESeR, which exists predominantly in the open xanthene form. This structural change resulted in drastic recovery of visible‐wavelength absorption and the ability to generate singlet oxygen (¹O₂). When HMDESeR‐βGal was applied to larval Drosophila melanogaster wing disks, which express β‐galactosidase only in the posterior region, photoirradiation induced cell death in the β‐galactosidase‐expressing region with high specificity.     

Investigation on the individual contributions of N?H···OC and C?H···OC interactions to the binding energies of β‐sheet models

In this article, the binding energies of 16 antiparallel and parallel β‐sheet models are estimated using the analytic potential energy function we proposed recently and the results are compared with those obtained from MP2, AMBER99, OPLSAA/L, and CHARMM27 calculations. The comparisons indicate that the analytic potential energy function can produce reasonable binding energies for β‐sheet models. Further comparisons suggest that the binding energy of the β‐sheet models might come mainly from dipole–dipole attractive and repulsive interactions and VDW interactions between the two strands. The dipole–dipole attractive and repulsive interactions are further obtained in this article. The total of N? H···H? N and C?O···O?C dipole–dipole repulsive interaction (the secondary electrostatic repulsive interaction) in the small ring of the antiparallel β‐sheet models is estimated to be about 6.0 kcal/mol. The individual N? H···O?C dipole–dipole attractive interaction is predicted to be ?6.2 ± 0.2 kcal/mol in the antiparallel β‐sheet models and ?5.2 ± 0.6 kcal/mol in the parallel β‐sheet models. The individual C^α? H···O?C attractive interaction is ?1.2 ± 0.2 kcal/mol in the antiparallel β‐sheet models and ?1.5 ± 0.2 kcal/mol in the parallel β‐sheet models. These values are important in understanding the interactions at protein–protein interfaces and developing a more accurate force field for peptides and proteins. © 2009 Wiley Periodicals, Inc. J Comput Chem 2010     

5‐(Alkynyl)dibenzothiophenium Triflates: Sulfur‐Based Reagents for Electrophilic Alkynylation

The synthesis of a series of 5‐(alkynyl) dibenzothiophenium triflates, prepared from dibenzo[b,d]thiophene 5‐oxide and the corresponding trimethylsilyl‐substituted alkynes is reported. Their structures were determined by X‐ray crystallography, and their reactivities as electrophilic alkynylation reagents evaluated. Their broad substrate scope and functional‐group tolerance illustrate their potential to become an alternative to the broadly used EBX reagents. Isotope labeling studies reveal that alkynyldibenzothiophenium salts may undergo attack by nucleophiles at either the α‐ or β‐carbon atom depending on the nature of their substitution pattern. Subsequent elimination of the dibenzothiophene unit and 1,2‐migration of one of the groups (in case of β‐attack) affords the desired alkynes.     

Building Proficient Enzymes with Foldamer Prostheses

Foldamers are non‐natural oligomers that adopt stable conformations reminiscent of those found in proteins. To evaluate the potential of foldameric subunits for catalysis, semisynthetic enzymes containing foldamer fragments constructed from α‐ and β‐amino acid residues were designed and characterized. Systematic variation of the α→β substitution pattern and types of β‐residue afforded highly proficient hybrid catalysts, thus demonstrating the feasibility of expanding the enzyme‐engineering toolkit with non‐natural backbones.