Mannose Units Research Articles

Synthesis and validation of clickable multimeric mannose ligands for dendritic cell targeting

The efficacy of therapeutics can be enhanced with their off-target effects being minimized through targeted delivery. In vaccination and cancer immunotherapy, targeting dendritic cells (DCs) and macrophages (Mφs) is crucial for the activation of the immune system. To this end, we have developed a set of carbohydrate-based ligands targeting C-type lectins (CTLs) present on the surface of DCs for receptor-mediated uptake. Branched structures functionalized with up to four mannose units were synthesized through CuI-catalysed click chemistry, and the products were further functionalized with the DBCO group to facilitate their conjugation with cargos of interest via Cu-free click chemistry. Thus, the ligands were conjugated with fluorescein azide, and the fluorescent conjugates were assessed for their internalization in CTL expressing immature DCs. The results showed that high-mannose branched structures were efficiently internalized through the endocytic pathway, with a positive qualitative correlation between branching degree of sugar-based ligands and internalization. As evidence for payload delivery, conjugating with a mannotriose ligand quantitatively increased the uptake of a fluorescently labeled oligonucleotide by immature DCs. Our work supports the use of click chemistry to synthesize and conjugate promising ligands for efficient delivery of nucleic acid-based drugs.

Synthesis of mannose conjugated biodegradable polyester-based nanocarriers and their binding study with Concanavalin A

In this paper, we aim to create fully biodegradable glyconanoparticles formed via self-assembly of glycopolymers, comprising of biodegradable aliphatic polyester conjugated with mannose moiety. To accomplish the goal, a series of random copolymers comprised of poly(propargyl glycolide-co-lactide) were synthesized. Alkyne moiety of the poly (propargyl glycolide) component was then clicked with mannose ethyl azide to produce amphiphilic glycopolymers in good yield (60–75 %). The glycopolymers were then self-assembled to form glyconanoparticles of 15–23 nm size in dry state and 78–88 nm size in hydrated state (hydrodynamic diameter). The copolymers were characterized by NMR and FTIR, whereas the nanoparticles were thoroughly characterized by DLS, FESEM, and HR-TEM and explored for their lectin binding efficiency. Isothermal calorimetry (ITC) experiments suggest a stronger binding efficiency of glyconanoparticles towards mannose-specific lectin such as Concanavalin A, as compared to its corresponding glycopolymers (∼2 fold) and monomeric mannose unit (∼7-fold) as well. Moreover, curcumin was selected as the model drug to be encapsulated (∼76 % encapsulation efficiency) and released (74 % in 24 h) from the glyconanoparticles. Apart from these, excellent haemocompatibility, cell viability, and cellular uptake of the nanoparticles (more than 80 % cells showed uptake of the nanoparticles), further supported their potential as drug carriers having sugar as a targeting moiety.

Recent Advances in Biomedical Applications of Mannans and Xylans.

Plant-based phytochemicals, including flavonoids, alkaloids, tannins, saponins, and other metabolites, have attracted considerable attention due to their central role in synthesizing nanomaterials with various biomedical applications. Hemicelluloses are the second most abundant among naturally occurring heteropolymers, accounting for one-third of all plant constituents. In particular, xylans, mannans, and arabinoxylans are structured polysaccharides derived from hemicellulose. Mannans and xylans are characterized by their linear configuration of β-1,4-linked mannose and xylose units, respectively. At the same time, arabinoxylan is a copolymer of arabinose and xylose found predominantly in secondary cell walls of seeds, dicotyledons, grasses, and cereal tissues. Their widespread use in tissue engineering, drug delivery, and gene delivery is based on their properties, such as cell adhesiveness, cost-effectiveness, high biocompatibility, biodegradability, and low immunogenicity. Moreover, it can be easily functionalized, which expands their potential applications and provides them with structural diversity. This review comprehensively addresses recent advances in the field of biomedical applications. It explores the potential prospects for exploiting the capabilities of mannans and xylans in drug delivery, gene delivery, and tissue engineering.

Well-Defined Oligo(azobenzene-graft-mannose): Photostimuli Supramolecular Self-Assembly and Immune Effect Regulation.

The immune system can recognize and respond to pathogens of various shapes. Synthetic materials that can change their shape have the potential to be used in vaccines and immune regulation. The ability of supramolecular assemblies to undergo reversible transformations in response to environmental stimuli allows for dynamic changes in their shapes and functionalities. A meticulously designed oligo(azobenzene-graft-mannose) was synthesized using a stepwise iterative method and "click" chemistry. This involved integrating hydrophobic and photoresponsive azobenzene units with hydrophilic and bioactive mannose units. The resulting oligomer, with its precise structure, displayed versatile assembly morphologies and chiralities that were responsive to light. These varying assembly morphologies demonstrated distinct capabilities in terms of inhibiting the proliferation of cancer cells and stimulating the maturation of dendritic cells. These discoveries contribute to the theoretical comprehension and advancement of photoswitchable bioactive materials.

N-glycan profiling of tissue samples to aid breast cancer subtyping

Breast cancer is a highly heterogeneous disease. Its intrinsic subtype classification for diagnosis and choice of therapy traditionally relies on the presence of characteristic receptors. Unfortunately, this classification is often not sufficient for precise prediction of disease prognosis and treatment efficacy. The N-glycan profiles of 145 tumors and 10 healthy breast tissues were determined using Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry. The tumor samples were classified into Mucinous, Lobular, No-Special-Type, Human Epidermal Growth Factor 2 + , and Triple-Negative Breast Cancer subtypes. Statistical analysis was conducted using the reproducibility-optimized test statistic software package in R, and the Wilcoxon rank sum test with continuity correction. In total, 92 N-glycans were detected and quantified, with 59 consistently observed in over half of the samples. Significant variations in N-glycan signals were found among subtypes. Mucinous tumor samples exhibited the most distinct changes, with 28 significantly altered N-glycan signals. Increased levels of tri- and tetra-antennary N-glycans were notably present in this subtype. Triple-Negative Breast Cancer showed more N-glycans with additional mannose units, a factor associated with cancer progression. Individual N-glycans differentiated Human Epidermal Growth Factor 2 + , No-Special-Type, and Lobular cancers, whereas lower fucosylation and branching levels were found in N-glycans significantly increased in Luminal subtypes (Lobular and No-Special-Type tumors). Clinically normal breast tissues featured a higher abundance of signals corresponding to N-glycans with bisecting moiety. This research confirms that histologically distinct breast cancer subtypes have a quantitatively unique set of N-glycans linked to clinical parameters like tumor size, proliferative rate, lymphovascular invasion, and metastases to lymph nodes. The presented results provide novel information that N-glycan profiling could accurately classify human breast cancer samples, offer stratification of patients, and ongoing disease monitoring.

Big is not better: Comparing two alpha-Gal-bearing glycotopes in neoglycoproteins as biomarkers for Leishmania (Viannia) braziliensis infection

The protozoan parasite Leishmania (Viannia) braziliensis is among Latin America's most widespread Leishmania species and is responsible for tegumentary leishmaniasis (TL). This disease has multiple clinical presentations, with cutaneous leishmaniasis (CL) being the most frequent. It manifests as one or a few localized skin ulcers, which can spread to other body areas. Hence, early diagnosis and treatment, typically with pentavalent antimonials, is critical. Traditional diagnostic methods, like parasite culture, microscopy, or the polymerase chain reaction (PCR) for detection of the parasite DNA, have limitations due to the uneven distribution of parasites in biopsy samples. Nonetheless, studies have revealed high levels of parasite-specific anti-α-Gal antibodies in L. (V.) braziliensis-infected patients. Previously, we demonstrated that the neoglycoprotein NGP28b, consisting of the L. (Leishmania) major type-2 glycoinositolphospholipid (GIPL)-3-derived trisaccharide Galpα1,6Galpα1,3Galfβ conjugated to bovine serum albumin (BSA) via a linker, acts as a reliable serological biomarker (BMK) for L. (V.) braziliensis infection in Brazil. This indicates the presence of GIPL-3 or a similar structure in this parasite, and its terminal trisaccharide either functions as or is part of an immunodominant glycotope. Here, we explored whether extending the trisaccharide with a mannose unit would enhance its efficacy as a biomarker for the serological detection of L. (V.) braziliensis. We synthesized the tetrasaccharide Galpα1,6Galpα1,3Galfβ1,3Manpα(CH2)3SH (G31SH) and conjugated it to maleimide-functionalized BSA to afford NGP31b. When we assessed the efficacy of NGP28b and NGP31b by chemiluminescent enzyme-linked immunosorbent assay on a cohort of CL patients with L. (V.) braziliensis infection from Bolivia and Argentina against a healthy control group, both NGPs exhibited similar or identical sensitivity, specificity, and accuracy. This finding implies that the mannose moiety at the reducing end is not part of the glycotope recognized by the parasite-specific anti-α-Gal antibodies in patients' sera, nor does it exert a relevant influence on the terminal trisaccharide's conformation. Moreover, the mannose does not seem to inhibit glycan-antibody interactions. Therefore, NGP31b is a viable and dependable BMK for the serodiagnosis of CL caused by L. (V.) braziliensis.

Advancing Antiamyloidogenic Activity by Fine-Tuning Macromolecular Topology.

Amyloid β peptide can aggregate into thin β-sheet fibrils or plaques deposited on the extracellular matrix, which is the hallmark of Alzheimer's disease. Multifunctional macromolecular structures play an important role in inhibiting the aggregate formation of amyloidogenic materials and thus are promising candidates with antiamyloidogenic characteristics for the development of next-generation therapeutics. In this study, we evaluate how small differences in the dendritic topology of these structures influence their antiamyloidogenic activity by the comparison of "perfectly dendritic" and "pseudodendritic" macromolecules, both decorated with mannose units. Their compactness, the position of surface units, and the size of glyco-architectures influence their antiamyloidogenic activity against Aβ 40, a major component of amyloid plaques. For the advanced analysis of the aggregation of the Aβ peptide, we introduce asymmetric flow field flow fractionation as a suitable method for the quantification of large and delicate structures. This alternative method focuses on the quantification of complex aggregates of Aβ 40 and glycodendrimer/glyco-pseudodendrimer over different time intervals of incubation, showing a good correlation to ThT assay and CD spectroscopy results. Kinetic studies of the second-generation glyco-pseudodendrimer revealed maximum inhibition of Aβ 40 aggregates, verified with atomic force microscopy. The second-generation glyco-pseudodendrimer shows the best antiamyloidogenic properties confirming that macromolecular conformation in combination with optimal functional group distribution is the key to its performance. These molecular properties were validated and confirmed by molecular dynamics simulation.

Glycolipids mimicking biosurfactants of the synthetic origin as new immunomodulating and anticandidal derivatives

The immunobiological effectivity of glycolipids mimicking biosurfactants of the synthetic origin was followed up using macrophages cell line RAW264.7. These derivatives with different number of mannose units connected glycosidically or through triazole linker, and all having octyl aglycone, were evaluated with respect to their structure - immunomodulation activity relationship. This comparative study showed that the structural variations of the selected derivatives influenced the immunobiological cell behaviour as concerned pro-inflammatory TNF-α, IL-6, IL-1α, IL-17, IL-12 and anti-inflammatory IL-10 cytokines production and enhancement of RAW264.7 cell proliferation. The derivatives with mannose units linked through triazole linkers exerted in some cases stronger immunomodulative potency than (di)mannosides. On the other hand, a presence of triazole linker is a less favourable for an effective candidacidal activity as determined by in vitro using Candida albicans biofilm. The design of new defined immunomodulating formulas of the synthetic origin as possible antifungal agents and prospective participants in drug delivery systems may be of interest.

Evaluation of Acemannan in Different Commercial Beverages Containing Aloe Vera (Aloe barbadensis Miller) Gel.

Aloe vera (Aloe barbadensis Miller) gel is a frequently used ingredient in many food pro-ducts, particularly beverages, due to its reported health benefits. Studies have identified acemannan, a polysaccharide rich in mannose units which are partially or fully acetylated, as the primary bioactive compound in Aloe vera gel. The acemannan content and its degree of acetylation (DA) were measured in 15 different commercial beverages containing Aloe vera at varying concentrations (from 30% to 99.8%) as listed on the label. Other biopolymers such as pectins, hemicelluloses, and cellulose were also evaluated. Flavoured beverages (seven samples labelled as containing from 30% to 77% Aloe vera) presented low levels of acemannan (<30 mg/100 g of fresh sample) and were fully deacetylated in most cases. These samples had high levels of other polymers such as pectins, hemicelluloses, and cellulose, likely due to the addition of fruit juices for flavour. Unflavoured beverages (eight samples, with Aloe vera concentrations above 99% according to their labels) had variable levels of acemannan, with only three containing more than 160 mg/100 g of fresh sample. In fact, four samples had less than 35 mg acemannan/100 g of fresh sample. DA levels in all but one sample were lower than 35%, possibly due to processing techniques such as pasteurization causing degradation and deacetylation of the acemannan polymer. Legislation regarding Aloe vera products is limited, and manufacturers are not required to disclose the presence or quality of bioactive compounds in their products, leaving consumers uncertain about the true properties of the products they purchase.

The Application of HPLC-FLD and NMR in the Monitoring of Therapy Efficacy in Alpha-Mannosidosis.

Alpha-mannosidosis is a rare lysosomal storage disorder, caused by decreased activity of α-D-mannosidase. This enzyme is involved in the hydrolysis of mannosidic linkages in N-linked oligosaccharides. Due to the mannosidase defect, undigested mannose-rich oligosaccharides (Man2GlcNAc - Man9GlcNAc) accumulating in cells are excreted in large quantities in urine. In this work, we determined the levels of urinary mannose-rich oligosaccharides in a patient subjected to novel enzyme replacement therapy. Urinary oligosaccharides were extracted using solid phase extraction (SPE), labeled by fluorescent tag 2-aminobenzamide, and quantified by high-performance liquid chromatography (HPLC) with fluorescence detector (FLD). The identity of peaks was determined by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry. In addition, the levels of urinary mannose-rich oligosaccharides were also quantified by 1H nuclear magnetic resonance (NMR) spectroscopy. The data were analyzed using one-tailed paired t-test and Pearson's correlation tests. Compared to levels before the administration of therapy, an approximately two-folds decrease in total mannose-rich oligosaccharides after one month of treatment was observed by NMR and HPLC. After four months, an approximately ten-folds significant decrease in total urinary mannose-rich oligosaccharides was detected, suggesting therapy effectiveness. A significant decrease in the levels of oligosaccharides with 7-9 mannose units was detected by HPLC. The application of both HPLC-FLD and NMR in quantification of oligosaccharide biomarkers is a suitable approach for monitoring of therapy efficacy in alpha-mannosidosis patients.

Introducing carbohydrate patterning in mannose-decorated supramolecular assemblies and hydrogels.

Benzene-1,3,5-tricarboxamide (BTA) glyco-monomers containing one, two or three mannose units are synthesized and formulated into differently patterned supramolecular glycopolymers through homo-assembly or co-assembly with non-functionalized BTAs. Unfortunately, no cellular activity could be detected. Excitingly, these glyco-BTA monomers could be formulated into hydrogels, paving the way for (immune) cell culture.

Mannose-functionalized star polycation mediated CRISPR/Cas9 delivery for lung cancer therapy.

The survivin gene, highly expressed in most cancer cells, is closely associated with inhibiting apoptosis. Therefore, gene editing for the survivin gene has great potential in tumor therapy. However, it is difficult for plasmid DNA (pDNA) to be taken up directly by cells, and thus the construction of gene vectors is a key step for successful gene editing. Ethanolamine-functionalized polyglycidyl methacrylate (PGEA) has been proved to facilitate the transfection of pDNA into cells in both in vivo and in vitro experiments. However, PGEA does not specifically recognize tumor cells. Some tumor cells express more mannose receptor (MR) than healthy cells. To achieve efficient target and transfection, we designed mannose-functionalized four-arm PGEA cationic polymers (P(GEA-co-ManMA), GM) with different molecular weights. GM was combined with pCas9-survivin. The mannose unit of GM/pCas9-survivin was identified by MR to selectively enter lung cancer cells. In vitro experiments showed that GM not only had excellent biocompatibility, gene transfection performance, and targeted ability, but also significantly inhibited the proliferation of tumor cells when used in combination with pCas9-survivin. At the same time, we also studied the relationship between the molecular weight and therapeutic effect.

Understanding the Structure and Rheology of Galactomannan Solutions with Coarse-Grained Modeling

Understanding and predicting the behavior of aqueous solutions of galactomannans is of significant interest for a variety of fields and applications, such as hydraulic fracturing in the petroleum industry and thickening in the food industry. Herein, we devise and validate a coarse-grained (CG) model of guar gum─a galactomannan─and study its structure and rheology in aqueous solution. The CG model is computationally efficient, utilizing an implicit solvent and one CG bead per monosaccharide, and exhibits good quantitative agreement with experiments with respect to the radius of gyration of the chain, aggregate structure in solution, and solution viscosity. Using both simulations and experiments, we show that the behavior of guar gum in water can be explained by scaling theory for polymers in good solvents. Namely, the radius of gyration of the chain shows a self-avoiding random walk behavior, and the solution specific viscosity ηsp exhibits the expected power-law scaling with the reduced concentration in different solution regimes (i.e., ηsp ∼ (c[η])1.3 in semidilute and ηsp ∼ (c[η])3.9 in entangled solutions). By extending our CG model to other galactomannans with different mannose-to-galactose ratios (M/G ratios) and different arrangements of unsubstituted mannose units, we show that high M/G ratios and long blocks of unsubstituted mannose units are responsible for aggregate formation in galactomannan solutions, which can reduce solution viscosity moderately.

Synthesis of antimicrobial polymers with mannose residues as binders for the FimH adhesin of Escherichia coli

AbstractRecently, antimicrobial polymers have been developed as novel antibiotics, and they are beneficial for human health. An advantage of these polymers is that they can be easily functionalized via the addition or modification of monomers. In this study, to enhance the bacterial selectivity and antimicrobial activity of a polymer, mannose monomers were copolymerized with aminopropyl monomers and isoalkyl monomers. These mannose monomers were found to mimic the mannan and N‐glycan moieties on mammalian cells interacting with the FimH adhesin of Escherichia coli. A mannose monomer with a short ethyl linker did not enhance the antimicrobial activity (minimum inhibitory concentration (MIC): 32 μg/ml) against E. coli; however, that with a longer hexyl linker enhanced the antimicrobial activity (MIC: 16 μg/ml) against E. coli selectivity (no effect on Staphylococcus aureus was observed). Further, the polymer containing the hexyl mannose residue did not develop resistance in E. coli after 15 passages. Consequently, polymers with mannose units, which have a high affinity for the FimH adhesin, can serve as effective and selective antimicrobial agents against E. coli.

Composition and Structure of Aspen (Pópulus trémula) Hemicelluloses Obtained by Oxidative Delignification.

In this study, hemicelluloses of aspen wood (Pópulus trémula) were obtained by oxidative delignification in an acetic acid-water-hydrogen peroxide medium at temperatures of 70-100 °C and a process time of 1-4 h. The maximum polysaccharide yield of up to 9.68 wt% was reported. The composition and structure of the hemicelluloses were studied using a complex of physicochemical methods: gas and gel permeation chromatography, Fourier-transform infrared spectroscopy, 2D nuclear magnetic resonance spectroscopy, and thermogravimetric analysis. The xylose, mannose, galactose, and glucose monomer units were identified in the hemicelluloses by gas chromatography. The weight average molecular weight Mw of the products determined by gel permeation chromatography was found to range within 8932-33,142 g/mol. The reported Fourier-transform spectra of the hemicelluloses contain all the bands characteristic of heteropolysaccharides; a weak lignin absorption signal in the spectra at 1500-1510 cm-1 is attributed to a minor content of phenolic fragments in the structure of the obtained hemicelluloses. The use of thermogravimetric analysis established that the hemicelluloses isolated from aspen wood are resistant against heating to temperatures of up to 90-100 °C and, upon further heating up to 400 °C, start destructing at an increasing rate. The antioxidant activity of the hemicelluloses was examined using the compounds that mimic free radicals (1,1-diphenyl-2-picrylhydrazyl) and hydroxyl radicals (salicylic acid). It was found that the activity of all polysaccharides in neutralizing DPPH and hydroxyl radicals is lower than the absorption capacity of vitamin C at all the tested concentrations (0.5, 2, and 5 mg/mL) and attains 81.7 and 82.9%, respectively.

High-Throughput Human Complement C3 N-Glycoprofiling Identifies Markers of Early Onset Type 1 Diabetes Mellitus in Children

Recently, it was shown that children at the onset of type 1 diabetes (T1D) have a higher proportion of oligomannose glycans in their total plasma protein N-glycome compared to their healthy siblings. The most abundant complement component, glycoprotein C3, contains two N-glycosylation sites occupied exclusively by this type of glycans. Furthermore, complement system, as well as C3, was previously associated with T1D. It is also known that changes in glycosylation can modulate inflammatory responses, so our aim was to characterize the glycosylation profile of C3 in T1D. For this purpose, we developed a novel high-throughput workflow for human C3 concanavalin A lectin affinity enrichment and subsequent LC-MS glycopeptide analysis which enables protein-specific N-glycosylation profiling. From the Danish Childhood Diabetes Register, plasma samples of 61 children/adolescents newly diagnosed with T1D and 84 of their unaffected siblings were C3 N-glycoprofiled. Significant changes of C3 N-glycan profiles were found. T1D was associated with an increase in the proportion of unprocessed glycan structures with more mannose units. A regression model including C3 N-glycans showed notable discriminative power between children with early onset T1D and their healthy siblings with area under curve of 0.879. This study confirmed our previous findings of plasma high-mannose glycan changes in a cohort of recent onset T1D cases, suggesting the involvement of C3 N-glycome in T1D development. Our C3 glycan-based discriminative model could be valuable in assessment of T1D risk in children.

Specific Binding of Ligand-Functionalized Thermoresponsive Microgels: Effect of Architecture, Ligand Density, and Hydrophobicity.

The biomolecular interaction of ligand-presenting switchable microgels is studied with respect to the polymer type, composition, and structure of the microgels. Monodisperse microgels are prepared through precipitation polymerization of N-isopropylacrylamide (PNIPAM microgels) or oligo(ethylene glycol methacrylamide)s (POEGMA microgels) in the presence of crosslinkers or in their absence (self-crosslinked). Functionalization with mannose or biotin as model ligands and affinity measurements upon heating/cooling are conducted to obtain mechanistic insights into how the microgel phase transition affects the specific interactions. In particular, we are interested in adjusting the crosslinking, swelling degree, and ligand density of mannose-functionalized microgels to reversibly catch and release mannose binding Escherichia coli by setting the temperature below or above the microgels' volume phase transition temperature (VPTT). The increased mannose density for collapsed microgels above the VPTT results in stronger E. coli binding. Detachment of E. coli by reswelling the microgels below the VPTT is achieved only for self-crosslinked microgels showing a stronger decrease in ligand density compared to microgels with dedicated crosslinkers. Owing to a reduced mannose density in the shell of POEGMA microgels, their E. coli binding was lower compared to PNIPAM microgels, as supported by ultraresolution microscopy. Importantly, an inverse temperature-controlled binding of microgels decorated with hydrophilic mannose and hydrophobic biotin ligands is observed. This indicates that hydrophobic ligands are inaccessible in the collapsed hydrophobic network above the VPTT, whereas hydrophilic mannose units are then enriched at the microgel-water interface and thus are more accessible.

(Invited, Digital Presentation) Polyaromatic Micelles: Discrete Nanoparticles with Versatile Host Functions in Water

Polyaromatic micelles, assembled from bent anthracene dimers with hydrophilic units, have been established by our group as versatile host structures in water at the nanoscale (Fig. 1a; M. Yoshizawa et al., Acc. Chem. Res. 2019, 52, 2392). We here report recent efforts into the direction of bioapplications of these micelles via deliberate modifications of the amphiphile design. Utilization of an ortho-dianthrylbenzene core is shown to allow construction of photoresponsive micelles capable of quantitative guest release via light irradiation, which is potentially useful for drug delivery (Fig. 1b). In addition, attachment of hydrophilic mannose units to the anthracene dimer framework is demonstrated to yield highly fluorescent saccharide clusters capable of selective interactions with proteins (Fig. 1c). Figure 1

Structural features of biologically active extracellular polysaccharide produced by green microalgae Dictyosphaerium chlorelloides

Ion-exchange chromatography of the biologically active extracellular biopolymer produced by D. chlorelloides yielded ten fractions differing in yield, protein content, monosaccharide composition and molecular weight distribution. Their sugar compositional analyses showed rhamnogalactans, substituted to different extent by mannose and glucose, as a dominant EPS component in all fractions (91 %) except one containing arabinogalactan (7 %). In highly branched rhamnogalactans the quantity of linear (1,3-; 1,4- and 1,6-linked) and branched β-D-galactose units (1,3,6-, 1,4,6- and 1,3,4,6-linked) was nearly equal. From various α-L-rhamnose linkages the 1,2,4-linkage was dominant. Data indicate a rhamnogalactan backbone of EPS, branched by terminal mannose and glucose units, and a lot of O-methylated derivatives of galactose residues (2-O-methyl, 2,3-O-dimethyl, 3-O-methyl and 6-O-methyl). In individual fractions their content and type varied. Detail study of the arabinogalactan showed that its backbone consists of 1,3-linked β-D-Galp units; some of them are branched through O-4 by 6-OMe-α-D-Galp- (1 → 2) -α-L-Araf side chain, other through O-6 by 3-OMe-β-D-Galp, 6-OMe-β-D-Galp, β-D-Galp and β-D-Galf.

Topological and Multivalent Effects in Glycofullerene Oligomers as EBOLA Virus Inhibitors.

The synthesis of new biocompatible antiviral materials to fight against the development of multidrug resistance is being widely explored. Due to their unique globular structure and excellent properties, [60]fullerene-based antivirals are very promising bioconjugates. In this work, fullerene derivatives with different topologies and number of glycofullerene units were synthesized by using a SPAAC copper free strategy. This procedure allowed the synthesis of compounds 1–3, containing from 20 to 40 mannose units, in a very efficient manner and in short reaction times under MW irradiation. The glycoderivatives were studied in an infection assay by a pseudotyped viral particle with Ebola virus GP1. The results obtained show that these glycofullerene oligomers are efficient inhibitors of EBOV infection with IC50s in the nanomolar range. In particular, compound 3, with four glycofullerene moieties, presents an outstanding relative inhibitory potency (RIP). We propose that this high RIP value stems from the appropriate topological features that efficiently interact with DC-SIGN.