Improving the Sensitivity for Quantifying Heparan Sulfate from Biological Samples.

Heparan sulfates (HSs) are widely expressed glycans in the animal kingdom. HS plays a role in regulating cell differentiation/proliferation, embryonic development, blood coagulation, inflammatory response, and viral infection. The amount of HS and its structural information are critically important for investigating the functions of HS in vivo. A sensitive and reliable quantitative technique for the analysis of HS from biological samples is under development. Here, we report a new labeling reagent for HS disaccharides analysis, 6-amino-N-(2-diethylamino)ethyl quinoline-2-carboamide (AMQC). The AMQC-conjugated disaccharides are analyzed by LC-MS/MS in positive mode, significantly improving the sensitivity. The use of AMQC coupled with authentic 13C-labeled HS disaccharide internal standards empowered us to determine the amount and the disaccharide composition of the HS on a single histological slide. We used this method to profile the levels of HS in the plasma/serum and tissues/organs to assist the disease prognosis in two animal models, including the acetaminophen (APAP)-induced acute liver injury mouse model and the burn injury mouse model. The method may uncover the roles of HS contributing to the diseases as well as provide a potential new set of biomarkers for disease diagnosis and prognosis.

Thiocarbonyl as a Switchable Relay-Auxiliary Group in Carbohydrate Synthesis.

A multifunctional O-phenyl thiocarbonyl (O(C═S)OPh) group was introduced in glycosylation reactions. This auxiliary group exhibits three features (1) C6-long-range participation effect, (2) relay activation, and (3) switchable promoter-controlled carbonylation, which enables the facile synthesis of both 6-deoxy glucoside and 6-alcohol glucoside. In addition, we successfully quantified the extent of the C6-acyl participation effect and developed its application toward the α-trisaccharide motif.

Synthesis of d-Galactosamine and d-Allosamine Derivatives via a Microwave-Assisted Preparation of 1,6-Anhydroglucosamine.

We report a microwave-assisted intramolecular anomeric protection (iMAP) of glucosamine, which facilitates concise transformation of 1,6-anhydroglucosamine into 1,6-anhydrogalactosamine and 1,6-anhydroallosamine. The iMAP simultaneously obviates both the O1 and O6 protection, and the differentiation between O3 and O4 can be well-controlled by the N2 functionality because of the hydrogen bonding between N2 and O4. Epimerization of O4 afforded the galactosamine derivative and that of O3 yielded allosamine.

Direct synthesis of methyl phosphoramidates in carbohydrates.

A direct installation of a methyl phosphoramidate group by using methyl benzylphosphoramidochloridate into carbohydrates and amino acid is described. This one-step synthesis is efficient for both primary and secondary alcohols and exhibited excellent regioselectivity and functional group compatibility. Formation of a single diastereomer is observed in certain cases. The N-benzyl protecting group on methyl phosphoramidates is easily removed under mild conditions.

Chemoselective per-O-trimethylsilylation and homogeneous N-functionalisation of amino sugars.

A highly efficient CH3CN-promoted hexamethyldisilazane per-O-trimethylsilylation of amino sugars was developed. Its applications in homogenous N-functionalisation and a concise synthesis of glucosamine 6-phosphate are described.

Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides.

Living organisms employ glycans as recognition elements because of their large structural information density. Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. Thus, easy, rapid, and efficient methods for building-block acquisition are desirable. Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages.