Glycosylated tris-bipyridine ferrous complexes for probing a mechanism behind carbohydrate-carbohydrate interactions: Spatial carbohydrate packing of glycoclusters changes on additions of salts in carbohydrate- and anion-dependent manners

2,2′-Bipyridines containing two β-maltoside, β-lactoside, or β-isomaltoside appendages were prepared and successively complexed with ferrous ion to afford hexavalent glycoclusters having tris-bipyridine ferrous complex cores. Each of these metalloglycoclusters showed unique UV–vis and CD spectral changes upon addition of chloride, nitrate, and sulfate salts in carbohydrate- and anion-dependent manners. The results indicate that spatial carbohydrate packing of the metalloglycoclusters changes upon addition of these anions and that different anions stabilize different carbohydrate packings. Furthermore, the sulfates specifically enhance the rheological properties of aqueous solutions containing the metalloglycocluster containing β-lactoside appendages.     

Dendri-RAFTs: a second generation of cyclopeptide-based glycoclusters

Synthetic glycoclusters and their related biological applications have stimulated increasing interest over the last decade. As a prerequisite to discovering active and selective therapeuticals, the development of multivalent glycoconjugates with diverse topologies is faced with inherent synthetic and structural characterisation difficulties. Here we describe a new series of molecularly-defined glycoclusters that were synthesized in a controlled manner using a robust and versatile divergent protocol. Starting from a Regioselectively Addressable Functionalized Template (RAFT) carrier, either a polylysine dendritic framework or a second RAFT, then 16 copies of βGal, αMan, βLac or cancer-related Thomsen-Freidenreich (αTF) antigen were successively conjugated within the same molecule using oxime chemistry. We thus obtained a new generation of dendri-RAFTs glycoclusters with high glycosidic density and variable spatial organizations. These compounds displaying 16 endgroups were unambiguously characterized by NMR spectroscopy and mass spectrometry. Further biological assays between a model lectin from Canavalia ensiformis (ConA) and mannosylated glycoclusters revealed a higher inhibition potency than the tetravalent counterpart, in particular for the hexadecavalent polylysine skeleton. Together with the efficiency of the synthetic and characterisation processes, this preliminary biological study provided clear evidence of promising properties that make the second generation of cyclopeptide-based glycoclusters attractive for biomedical applications.     

Hyaluronan-based glycoclusters as probes for chemical glycobiology

A strategy is described for the synthesis of β-(1,3)-GlcA-GlcNAc dimeric and tetrameric glycoclusters through the conjugation of disaccharide groups onto a diaminodiamide aromatic scaffold by reductive amination.     

Probing carbohydrate-lectin recognition in heterogeneous environments with monodisperse cyclodextrin-based glycoclusters

A series of β-cyclodextrin (βCD)-scaffolded glycoclusters exposing heterogeneous yet perfectly controlled displays of α-mannosyl (α-Man) and β-lactosyl (β-Lact) antennas were synthesized to probe the mutual influence of varying densities of the saccharide motifs in the binding properties toward different plant lectins. Enzyme-linked lectin assay (ELLA) data indicated that the presence of β-Lact residues reinforced binding of α-Man to the mannose-specific lectin concanavalin A (Con A) even though homogeneous β-Lact clusters are not recognized at all by this lectin, supporting the existence of synergic recognition mechanisms (heterocluster effect). Conversely, the presence of α-Man motifs in the heteroglycoclusters also resulted in a binding-enhancing effect of β-Lact toward peanut agglutinin (PNA), a lectin strongly binding multivalent lactosides but having no detectable affinity for α-mannopyranosides, for certain architectural arrangements. Two-site, sandwich-type ELLA data corroborated the higher lectin clustering efficiency of heterogeneous glycoclusters compared with homogeneous displays of the putative sugar ligand with identical valency. A turbidity assay was also consistent with the previous observations. Most revealingly, the lectin cross-linking ability of heterogeneous glycoclusters was sensitive to the presence of high concentrations of the non-ligand sugar, strongly suggesting that "mismatching" saccharide motifs may modulate carbohydrate-lectin specific recognition in a lectin-dependent manner when present in highly dense displays together with the "matching" ligand, a situation frequently encountered in biological systems.     

Glycosylation of organic phosphorus thio- and selenoacids—II: The reaction of organic phosphorus thioselenoacids with glycosyl bromides under kinetically and thermodynamically controlled conditions

Ambident anions derived from phosphorus thioselcnoacids were glycosylated with 2,3,4,6 - tetra - O - acetyl - α - d - glucopyranosyl bromide, 2,3,4,6 - tetra - O - acetyl - α - d - galactopyranosyl bromide and 2,3,4 - tri- O - acetyl - α - d - xylopyranosyl bromide. The products were β-Se-glucosyl- and β-S-glucosylthioselenoates. The Se/S ratio of the glycosylated phosphorothioselenoates depends on the reaction conditions. At higher temperatures an equilibrium was observed. As a result of this equilibrium the Se/S ratio of the linkages formed in the glycosylated products was different from that observed under kinetic control. The structures of the glycosylated phosphorothioselenoates were confirmed by spectroscopy, independent synthesis and selective oxidation.     

Divergent behavior of glycosylated threonine and serine derivatives in solid phase Peptide synthesis

Solid phase peptide coupling of glycosylated threonine derivatives was systematically evaluated. In contrast to glycosylated serine derivatives which are highly prone to epimerization, glycosylated threonine derivatives produce only negligible amounts of epimerization. Under forcing conditions, glycosylated threonine analogs undergo β-elimination, rather than epimerization. Mechanistic studies and molecular modeling were used to understand the origin of the differences in reactivity.     

Synthesis of dodecavalent fullerene-based glycoclusters and evaluation of their binding properties towards a bacterial lectin

Multivalency is playing a major role in biological processes and particularly in lectin-carbohydrate interactions. The design of high-affinity ligands of lectins should provide molecules capable of interfering with these biological processes and potentially inhibit bacterial or viral infections. Azide-alkyne "click" chemistry was applied to the synthesis of dodecavalent fullerene-based glycoclusters. The conjugation could be efficiently performed from alkyne or azide functions on either partners (i.e. hexakis-fullerene adduct or glycoside). PA-IL is a bacterial lectin from the opportunistic pathogen Pseudomonas aeruginosa and is involved in the recognition of glycoconjugates on human tissues. The glycoclusters obtained were evaluated as ligands of PA-IL and for their potential for competing with its binding to glycosylated surfaces. The affinities measured by hemagglutination inhibition assay (HIA), enzyme-linked lectin assay (ELLA), and surface plasmon resonance (SPR) displayed a significant "glycoside cluster effect" with up to a 12,000-fold increase in binding when comparing a monovalent carbohydrate reference probe with a dodecavalent fullerene-based glycocluster, albeit with some differences depending on the analytical technique.     

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.     

Homogeneous human complex-type oligosaccharides in correctly folded intact glycoproteins: evaluation of oligosaccharide influence on protein folding, stability, and conformational properties

The N-glycosylation of proteins is generated at the consensus sequence NXS/T (where X is any amino acid except proline) by the biosynthetic process, and occurs in the endoplasmic reticulum and Golgi apparatus. In order to investigate the influence of human complex-type oligosaccharides on counterpart protein conformation, crambin and ovomucoide, which consist of 46 and 56 amino acid residues, respectively, were selected for synthesis of model glycoproteins. These small glycoproteins were intentionally designed to be glycosylated at the α-helix (crambin: 8?position), β-sheet (crambin: 2?position) and loop position between the antiparallel β-sheets (ovomucoide: 28?position), and were synthesized by using a peptide-segment coupling strategy. After preparation of these glycosylated polypeptide chains, protein folding experiments were performed under redox conditions by using cysteine-cystine. Although the small glycoproteins bearing intentional glycosylation at the α-helix and β-sheet exhibited a suitable folding process, glycosylation at the loop position between the antiparallel β-strands caused multiple products. The conformational differences in the isolated homogeneous glycoproteins compared with non-glycosylated counterparts were evaluated by circular dichroism (CD) and NMR spectroscopy. These analyses suggested that this intentional N-glycosylation did not result in large conformational changes in the purified protein structures, including the case of glycosylation at the loop position between the antiparallel β-strands. In addition to these experiments, the conformational properties of three glycoproteins were evaluated by CD spectroscopy under different temperatures. The oligosaccharides on the protein surface fluctuated considerably; this was dependent on the increase in the solution temperature and was thought to disrupt the protein tertiary structure. Based on the measurement of the CD spectra, however, the glycoproteins bearing three disulfide bonds did not exhibit any change in their protein tertiary structure. These results suggest that the oligosaccharide conformational fluctuations were not disruptive to protein tertiary structure, and the tertiary structure of glycoproteins might be stabilized by the disulfide bond network.     

Design and Synthesis of Glycosylated Aromatic Nitrogen Mustard Derivatives

Antibody-directed enzyme prodrug therapy(ADEPT) is a new strategy for the treatment of cancer that has arisen in recent twenty years, the main merits of which are that it can improve the selectivity of anticancer drugs and reduce the side effects in remote tissue. In the present study, two prodrugs-glycosylated aromatic nitrogen mustard derivatives were synthesized. Glucose and lactose were converted into glycosyl donors-trichloroacetimidate; the obtained glycosyl donors were glycosylated with p-nitrophenol(glycosyl donors) to form β-glucosyl p-nitrobenzene and β-lactosyl p-nitrobenzene that were protected by acetyl in a stereoselective manner; the two products were reduced by zinc dust and then treated with ethylene oxide, afforded two glycosylated nitrogen mustard derivatives that were protected by acetyl; the last step was to deacetylate and then afforded the two target compounds that could be used as prodrugs of ADEPT for further Anti-tumor research.     

Formation of 3Pseudorotaxanes from β-cyclodextrin and a Series of Dicarboxylic Acids with Their Corresponding α,ω-alkanedicarboxylate Anions

Inclusion complexes between β-cyclodextrin (β-CD) and a series of dicarboxylic acids (DAn, n=11-15) were prepared by co-grinding and co-precipitation methods and the 3pseudorotaxane structure of them was eluci-dated by FTIR, DTA and XRD characterizations. Inclusion complexes of β-CD and α,w-alkanedicarboxylate anions (DAn2-) were acquired by neutralizing β-CD/DAn different inclusion complexes with sodium hydrox-ide and the structure was also proved to be a pseudorotaxane structure by 1H-NMR spectra and NOESY spectrum. Both the inclusion complexes of β-CD/DAn and β-CD/DAn2- adopt the 3pseudorotaxane structure with β-CD arranged in dimers threaded onto one aliphatic chain and the binding mode of 1:1 inclusion complex was excluded based on the consideration of chain conformations     

Modelling of molecular interactions and inclusion phenomena in substituted β-cyclodextrin: From simple probes to proteins

The ability of β-cyclodextrin (βCD) to form stable complexes with α-interferon was investigated. By using simple molecular mechanics approach interaction energy profiles of simple probes passing the center of βCD ring cavity along the main molecular symmetry axis were evaluated first. A computational study of host-guest inclusion complexes between βCD and L-α-aminoacids and some selected pentapeptides was also carried out and aimed at understanding the nature of the driving forces and mechanism, leading to their formation. Relative complexation energies for the complexes and the solvation Gibbs free energies for single L-α-aminoacids were calculated. Both the aminoacid residue inside the βCD cavity and neighbouring residues were found to contribute to the stabilization of βCD complexes with the side-chain of aminoacids present on the surface of α-interferon. The most appropriate number of host βCD molecules for the encapsulation in the first shell of one α-interferon molecule resulted to be 25.     

Glycosylation of organic thio- and selenoacids of phosphorus : The reaction of organic monothio acids of phosphorus with glycosyl bromides. Thiono-thiolo rearrangement of glycosyl phosphorothioates

Ambident anions derived from phosphorus monothio acids have been glycosylated by 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl-, 2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl- and 2,3,4-tri-acetyl-α-D-xylopyranosyl bromides, yielding β-S-glycosyl- and /gb-O-glycosylthioates. The S/O ratio of the glycosylated phosphorothioates depends markedly on the salt used. Ammonium salts favour the formation of S-derivatives, whereas silver salts give mainly O-derivatives. O-Glucosyl phosphorothioate undergoes thermal isomerisation to S-glucosyl phosphorothioate, with retention of configuration at the glycosylic centre. The implications of this stereochemical result are briefly discussed.     

Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites

Determination of O-glycosylation sites in glycopeptides was developed by using two model compounds designed from mucin2 tandem repeat motif and erythropoietin. β-Elimination/addition reaction using dimethylamine on glycosylated site through a Michael-type condensation produced efficient deglycosylation with appropriate chemical modification. The use of dimethylamine was efficient to release the O-linked glycan in a reaction time period of 2-6 h at 55 °C. Peptide sequencing was then performed using the liquid chromatography/quadrupole time-of-flight mass spectrometry and MS-MS experiments. Interpretation of fragmentation pathways of the β-elimination/addition products enabled straightforward recognition of glycosylation site. Compared to the fragmentation of corresponding native peptides, mass shift of −18 Da or +27 Da was clearly observed for the two kinds of β-elimination/addition products of the glycosylated threonine. Dimethylamine was found to provide higher efficiency of β-elimination/addition than methylamine and ammonia.     

基于环糊精二聚体与金刚烷修饰的温敏性聚合物的主客体识别构筑超分子水凝胶

利用环糊精二聚体(trans-Stilbene β-CD Dimer)与金刚烷修饰的温敏性聚合物(Pnipam-Ad)的主客体识别作用构筑了超分子水凝胶. 2D NMR 测定结果表明trans-Stilbene β-CD Dimer 和Pnipam-Ad 的主客体相互作用是通过β-CD 空腔和疏水体Ad 形成包结络合物进行的. 环糊精二聚体trans-Stilbene β-CD Dimer 和聚合物Pnipam-Ad 两者之间的缔合程度受trans-Stilbene β-CD Dimer 和Pnipam-Ad 的浓度以及trans-Stilbene β-CD Dimer 光异构的影响. 此外, 由于trans-Stilbene β-CD Dimer 对Pnipam-Ad 聚合物链的物理交联作用使两者混合溶液的最低临界溶解温度(LCST)低于纯聚异丙基丙烯酰胺(Pnipam)的LCST.     

Synthesis of l-arabinopyranose containing hederagenin saponins

The synthesis of eight hederagenin saponins, five of which are natural products, and their methyl esters is described as part of an ongoing study of the biological activity of triterpenoid saponins. Six disaccharides consisting of an l-arabinopyranose glycosylated in positions 2, 3, or 4 with a β-d-xylopyranose or a β-d-glucopyranose residue, respectively, were synthesized in good to excellent yields. The saponins were then prepared in good yields through glycosylation with a suitably protected hederagenin derivative followed by total deprotection and treatment with diazomethane.     

Solid-phase carbohydrate synthesis via on-bead protecting group chemistry

Di- and tri-saccharides were synthesized on a solid phase. The procedure started with a non-protected sugar linked via either cysteine or glutamine to a polystyrene resin. Selective dimethoxytritylation chemistry and subsequent steps yielded a resin-bound acceptor that could be glycosylated to yield β1,6-linked disaccharides. Reiteration of the procedure produced the trisaccharide.     

含哌啶取代酞菁金属配合物的合成及其对癌细胞光灭活作用

采用“模板法”合成8种含哌啶取代酞菁金属配合物[(PEO)4PcM, M=Zn, Ni, Co, Cu, PEO=2-(哌啶-1-基)乙氧基], 采用FTIR、质谱和元素分析等技术对其进行了表征. 分别测定了它们的紫外-可见吸收光谱、荧光发射光谱和光敏化产生单线态氧的能力. 研究结果表明, 2种酞菁锌配合物均具有较高的摩尔消光系数、一定的荧光量子产率和较大的单线态氧生成速率. 通过光动力灭活BEL7402肝癌细胞的试验研究发现, β-(PEO)4PcZn的浓度为10 μmol/L时, 在670 nm激光辐照下, 光剂量为1.2 J时, 对癌细胞的抑制率可达到83%.     

NO_2,OH和OH~-对环四甲撑四硝胺初始热解的影响

在密度泛函理论的(DFT)B3LYP/6-31g(d)水平上,优化得到了环四甲撑四硝胺(β-HMX)及其与高氯酸铵(AP)裂解产物NO2、OH及OH-分别形成复合物的各种稳定构型,计算了β-HMX及各复合物中最弱的N—NO2键解离能.结果发现:β-HMX与NO2、OH结合后构型变化不是很大,但对称性降低;β-HMX与OH-结合后,HMX构型发生较大变化,原有的对称性明显遭到破坏.计算表明:NO2易与HMX骨架环上亚甲基(—CH2—)中的H作用,"置换"出H而引发HMX的热解,从而改变了HMX的初始分解通道;OH对HMX的N—NO2键解离影响不大,而OH-与β-HMX结合后其N—NO2键解离能比β-HMX降低近200kJ.mol-1,表明OH-对其裂解有明显的促进作用.NO2、OH-的存在可使HMX的分解温度大大降低.     

Reaction of α,β-unsaturated Fischer carbene complexes with allyl alkoxide

Optically enriched homo-binuclear Fischer chromium carbene complexes with planar chiral arene chromium complexes gave α-allyl β-arylpropionates up to 97% ee by reaction with allyl alkoxide and subsequent photo-oxidative demetalation. The chiral hetero-binuclear tungsten carbene complexes afforded anti α-allyl β-hydroxy β-arylpropionates as a major product up to 92/8 dr by the same reaction sequence. High diastereoselectivity in these reactions is contributed to the planar chirality of the arene chromium complex, even though the reaction was carried out under vigorous basic media. The reaction products, α-allyl β-arylpropionates were derived by 1,3-M(CO)₅ shift and subsequent [3,3]-sigmatropic rearrangement. Also, the corresponding chromium-uncomplexed α,β-unsaturated Fischer carbene complexes afforded α-allyl β-arylpropionates under the same conditions. Formation of β-allyl β-arylpropionates via 1,2-M(CO)₅ shift followed by [3,4]-sigmatropic rearrangement was not observed in both reactions of chromium-coordinated and the corresponding chromium-uncoordinated α,β-unsaturated Fischer carbene complexes with allyl alkoxide in the presence of base.