Glycosidic Moiety Research Articles

Correlating In Vitro Target-Oriented Screening and Docking: Inhibition of Matrix Metalloproteinases Activities by Flavonoids.

Metalloproteases are a family of zinc-containing endopeptidases involved in a variety of pathological disorders. The use of flavonoid derivatives as potential metalloprotease inhibitors has recently increased.Particular plants growing in Sicily are an excellent yielder of the flavonoids luteolin, apigenin, and their respective glycoside derivatives (7-O-rutinoside, 7-O-glucoside, and 7-O-glucuronide).The inhibitory activity of luteolin, apigenin, and their respective glycoside derivatives on the metalloproteases MMP-1, MMP-3, MMP-13, MMP-8, and MMP-9 was assessed and rationalized correlating in vitro target-oriented screening and in silico docking.The flavones apigenin, luteolin, and their respective glucosides have good ability to interact with metalloproteases and can also be lead compounds for further development. Glycones are more active on MMP-1, -3, -8, and -13 than MMP-9. Collagenases MMP-1, MMP-8, and MMP-13 are inhibited by compounds having rutinoside glycones. Apigenin and luteolin are inactive on MMP-1, -3, and -8, which can be interpreted as a better selectivity for both -9 and -13 peptidases. The more active compounds are apigenin-7-O-rutinoside on MMP-1 and luteolin-7-O-rutinoside on MMP-3. The lowest IC50 values were also found for apigenin-7-O-glucuronide, apigenin-7-O-rutinoside, and luteolin-7-O-glucuronide. The glycoside moiety might allow for a better anchoring to the active site of MMP-1, -3, -8, -9, and -13. Overall, the in silico data are substantially in agreement with the in vitro ones (fluorimetric assay).

Dairy propionibacteria prevent the proliferative effect of plant lectins on SW480 cells and protect the metabolic activity of the intestinal microbiota in vitro

Plant lectins are specific carbohydrate-binding proteins that are widespread in legumes such as beans and pulses, seeds, cereals, and many plants used as farm feeds. They are highly resistant to cooking and digestion, reaching the intestinal lumen and/or blood circulation with biological activity. Since many legume lectins trigger harmful local and systemic reactions after their binding to the mucosal surface, these molecules are generally considered anti-nutritive and/or toxic substances. In the gut, specific cell receptors and bacteria may interact with these dietary components, leading to changes in intestinal physiology. It has been proposed that probiotic microorganisms with suitable surface glycosidic moieties could bind to dietary lectins, favoring their elimination from the intestinal lumen or inhibiting their interaction with epithelial cells. In this work, we assessed in vitro the effects of two representative plant lectins, concanavalin A (Con A) and jacalin (AIL) on the proliferation of SW480 colonic adenocarcinoma cells and metabolic activity of colonic microbiota in the absence or presence of Propionibacterium acidipropionici CRL 1198. Both lectins induced proliferation of colonic cells in a dose-dependent manner, whereas ConA inhibited fermentative activities of colonic microbiota. Pre-incubation of propionibacteria with lectins prevented these effects, which could be ascribed to the binding of lectins by bacterial cells since P. acidipropionici CRL 1198 was unable to metabolize these proteins, and its adhesion to colonic cells was reduced after reaction with Con A or AIL. The results suggest that consumption of propionibacteria at the same time as lectins could reduce the incidence of lectin-induced alterations in the gut and may be a tool to protect intestinal physiology.

Structure–activity relationships of fraxamoside as an unusual xanthine oxidase inhibitor

Fraxamoside, a macrocyclic secoiridoid glucoside featuring a hydroxytyrosol group, was recently identified as a xanthine oxidase inhibitor (XOI) comparable in potency in vitro to the standard antigout drug allopurinol. However, this activity and its considerably higher value than its derivatives oleuropein, oleoside 11-methyl ester, and hydroxytyrosol are not explained by structure–activity relationships (SARs) of known XOIs. To exclude allosteric mechanisms, we first determined the inhibition kinetic of fraxamoside. The resulting competitive mechanism prompted a computational SAR characterization, combining molecular docking and dynamics, which fully explained the behavior of fraxamoside and its derivatives, attributed the higher activity of the former to conformational properties of its macrocycle, and showed a substantial contribution of the glycosidic moiety to binding, in striking contrast with glycoside derivatives of most other XOIs. Overall, fraxamoside emerged as a lead compound for a new class of XOIs potentially characterized by reduced interference with purine metabolism.

Concanavalin A Coated Activated Carbon for High Performance Enzymatic Catalysis

A facile method of immobilizing enzymes on activated carbon (AC) is proposed, in which the first step is to coat AC with Concanavalin A (ConA), followed by the adsorption of enzymes. Two model glycoenzymes, horseradish peroxidase and laccase, were immobilized on the ConA adsorbed AC through the tightly specific recognition of ConA to their glycosidic moieties, as confirmed by laser confocal microscopy. The coating layer of ConA reduced the deactivation of enzymes and prevented the leakage of enzymes, as indicated by the significantly improved yield of enzymatic activity. The immobilized enzymes on ConA coated AC appeared an improved stability against pH and temperature, offering an expanded operation “window” for growing applications of the enzymatic catalysis. Finally, the integration of the adsorption capacity of AC and the chemical degradation of laccase promises the efficient removal of aqueous phenol. The improved recovery of enzyme activity, enhanced stability as well as the combination of substrate...

Chlorophyll Catabolites in Fall Leaves of the Wych Elm Tree Present a Novel Glycosylation Motif.

Fall leaves of the common wych elm tree (Ulmus glabra) were studied with respect to chlorophyll catabolites. Over a dozen colorless, non‐fluorescent chlorophyll catabolites (NCCs) and several yellow chlorophyll catabolites (YCCs) were identified tentatively. Three NCC fractions were isolated and their structures were characterized by spectroscopic means. Two of these, Ug‐NCC‐27 and Ug‐NCC‐43, carried a glucopyranosyl appendage. Ug‐NCC‐53, the least polar of these NCCs, was identified as the formal product of an intramolecular esterification of the propionate and primary glucopyranosyl hydroxyl groups of Ug‐NCC‐43. Thus, the glucopyranose moiety and three of the pyrrole units of Ug‐NCC‐53 span a 20‐membered ring, installing a bicyclo[17.3.1]glycoside moiety. This structural motif is unprecedented in heterocyclic natural products, according to a thorough literature search. The remarkable, three‐dimensional bicyclo[17.3.1]glycoside architecture reduces the flexibility of the linear tetrapyrrole. This feature of Ug‐NCC‐53 is intriguing, considering the diverse biological effects of known bicyclo[n.3.1]glycosidic natural products.

New Reported Flavonol Characterized by NMR from the Petals of <i>Talipariti elatum S. w.</i> in Cuba

From yellow petals of Blue Mahoe, besides the known flavonoids gossypetin and gossypitrin, a new one gossypetin derivative was isolated from ethanolic extracts after Soxhlet extraction. Hence, in this study, we present a validated, sensitive and reliable NMR method for the simultaneous identification of flavonoids in this flower drug. Structure analyses of this flavonoid, revealed the identical glycoside moiety attached to a flavonol skeleton like gossypitrin, for which the structure of gossypetin-3'-O-β-glucoside was deduced from extensive NMR experiments.

Synthesis of the tetrasaccharide glycoside moiety of Solaradixine and rapid NMR-based structure verification using the program CASPER

The major glycoalkaloid in the roots of Solanum laciniatum is Solaradixine having the branched tetrasaccharide β-d-Glcp-(1→2)-β-d-Glcp-(1→3)[α-l-Rhap-(1→2)]-β-d-Galp linked to O3 of the steroidal alkaloid Solasodine. We herein describe the synthesis of the methyl glycoside of the tetrasaccharide using a super-armed disaccharide as a donor molecule. A 2-(naphthyl)methyl protecting group was used in the synthesis of the donor since it was tolerant to a wide range of reaction conditions. The 6-O-benzylated-hexa-O-tert-butyldimethylsilyl-protected β-d-Glcp-(1→2)-β-d-Glcp-SEt donor, which avoided 1,6-anydro formation, was successfully glycosylated at O3 of a galactoside acceptor molecule. However, subsequent glycosylation at O2 by a rhamnosyl donor was unsuccessful and instead a suitably protected α-l-Rhap-(1→2)-β-d-Galp-OMe disaccharide was used as the acceptor molecule together with a super-armed β-d-Glcp-(1→2)-β-d-Glcp-SEt donor in the glycosylation reaction, to give a tetrasaccharide in a yield of 55%, which after deprotection resulted in the target molecule, the structure of which was verified by the NMR chemical shift prediction program CASPER.

Investigation of inclusion complexes of some anthocyanins with hydroxypropyl-β-cyclodextrin

In the present work, a spectrophotometric method (including electronic spectra differences) was applied to investigate complex formations between anthocyanins with hydroxypropyl-β-cyclodextrin (HPbCD). Also the molecular mechanics method (MM+) was used to analyze the system. The possibility of inclusion of complex formations of anthocyanin in flavylium form by incorporating the B ring into “host” cavity is predicted theoretically and is proved experimentally. Complex stability was shown to depend on steric factors in the complexes, defined by the glycoside moiety size at 3 position; due to the change of the closest surrounding geometry of carbon at 2 position (when hybridization of type changes sp2 → sp3), these stresses decrease, which allows the formation of more stable complexes between cyclodextrin and hemiacetal anthocyanin forms accompanied by solution bleaching. The slow anthocyanin forms equilibration is shown to be a property of the systems containing and missing cyclodextrin, and this effect is due to slow equilibration establishment of hemiacetal–chalkone transitions, whose role in inclusion complex formation is negligible.

Inhibition of Human UDP-Glucuronosyltransferase Enzymes by Canagliflozin and Dapagliflozin: Implications for Drug-Drug Interactions.

Canagliflozin (CNF) and dapagliflozin (DPF) are the first sodium-glucose cotransporter 2 inhibitors to be approved for clinical use. Although available evidence excludes clinically significant inhibition of cytochromes P450, the effects of CNF and DPF on human UDP-glucuronosyltransferase (UGT) enzymes are unknown. Here, we report the inhibition of human recombinant UGTs by CNF and DPF, along with the Ki values for selected recombinant and human liver microsomal UGTs. CNF inhibited all UGT1A subfamily enzymes, but the greatest inhibition was observed with UGT1A1, UGT1A9, and UGT1A10 (IC50 values ≤ 10 µM). DPF similarly inhibited UGT1A1, UGT1A9, and UGT1A10, with IC50 values ranging from 39 to 66 µM. In subsequent kinetic studies, CNF inhibited recombinant and human liver microsomal UGT1A9; Ki values ranged from 1.4 to 3.0 µM, depending on the substrate (propofol/4-methylumbelliferone) enzyme combination. Ki values for CNF inhibition of UGT1A1 were approximately 3-fold higher. Consistent with the activity screening data, DPF was a less potent inhibitor of UGT1A1 and UGT1A9. The Ki for DPF inhibition of UGT1A1 was 81 µM, whereas the Ki values for inhibition of UGT1A9 ranged from 12 to 15 µM. Based on the in vitro Ki values and plasma concentrations reported in the literature, DPF may be excluded as a perpetrator of DDIs arising from inhibition of UGT enzymes, but CNF inhibition of UGT1A1 and UGT1A9 in vivo cannot be discounted. Since the sodium-glucose cotransporter 2 inhibitors share common structural features, notably a glycoside moiety, investigation of drugs in this class for effects on UGT to identify (or exclude) potential drug-drug interactions is warranted.

Highly Stereoselective Synthesis of Natural‐Product‐Like Hybrids by an Organocatalytic/Multicomponent Reaction Sequence

AbstractIn an endeavor to provide an efficient route to natural product hybrids, described herein is an efficient, highly stereoselective, one‐pot process comprising an organocatalytic conjugate addition of 1,3‐dicarbonyls to α,β‐unsaturated aldehydes followed by an intramolecular isocyanide‐based multicomponent reaction. This approach enables the rapid assembly of complex natural product hybrids including up to four different molecular fragments, such as hydroquinolinone, chromene, piperidine, peptide, lipid, and glycoside moieties. The strategy combines the stereocontrol of organocatalysis with the diversity‐generating character of multicomponent reactions, thus leading to structurally unique peptidomimetics integrating heterocyclic, lipidic, and sugar moieties.

Highly stereoselective synthesis of natural-product-like hybrids by an organocatalytic/multicomponent reaction sequence.

In an endeavor to provide an efficient route to natural product hybrids, described herein is an efficient, highly stereoselective, one-pot process comprising an organocatalytic conjugate addition of 1,3-dicarbonyls to α,β-unsaturated aldehydes followed by an intramolecular isocyanide-based multicomponent reaction. This approach enables the rapid assembly of complex natural product hybrids including up to four different molecular fragments, such as hydroquinolinone, chromene, piperidine, peptide, lipid, and glycoside moieties. The strategy combines the stereocontrol of organocatalysis with the diversity-generating character of multicomponent reactions, thus leading to structurally unique peptidomimetics integrating heterocyclic, lipidic, and sugar moieties.

Structural characterization of flavonoid C- and O-glycosides in an extract of Adhatoda vasica leaves by liquid chromatography with quadrupole time-of-flight mass spectrometry.

Adhatoda vasica Nees is a well-known Ayurvedic medicinal plant, belonging to the family Acanthaceae. This study aims to seek identification and characterization of flavonoid C- and O-glycosides in the aqueous fraction of the plant leaves. A method was developed for simultaneous characterization of flavonoids and their glycosides using high-pressure liquid chromatography with quadrupole time-of-flight mass spectrometry (HPLC/ESI-QTOF-MS/MS). The chromatographic separation was carried on an Agilent Poroshell 120 EC-C18 column (4.6 × 150 mm, 2.7 µm) operated with 0.1% formic acid aqueous solution and methanol as the mobile phase. The fragmentations of the studied [M-H](-) ions of C-glycosides were shown to be cross-ring cleavages of the glycoside moiety [M-H-(60/90/120)](-) whereas O-glycosides were shown to eliminate the sugar moiety (Y0 (-) or [Y0 -H](-) ) from the aglycone unit; 6-C-glycosides exhibited [M-H-18](-) , a characteristic ion, and also a higher abundance of (0,3) X6 or 8 ions in comparison to 8-C glycosides; flavonoid 6,8-di-C-glycosides exhibited cross-ring cleavages of the sugar attached to the C-6 position preferentially. This method was successfully applied for analysis of flavonoids and their glycosides in Adhatoda vasica leaves. A total of 29 compounds were tentatively identified including 17 C-, nine O-glycosides and three flavonoids.

Surface-enhanced Raman scattering (SERS) study of anthocyanidins

Anthocyanins are an important class of natural compounds responsible for the red, purple and blue colors in a large number of flowers, fruits and cereal grains. They are polyhydroxy- and polymethoxy-derivatives of 2-phenylbenzopyrylium (flavylium) salts, which are present in nature as glycosylated molecules. The aim of the present study is to assess the identification of anthocyanidins, i.e. anthocyanins without the glycosidic moiety, by means of surface-enhanced Raman spectroscopy (SERS), a very chemically-specific technique which is moreover sensitive to subtle changes in molecular structures. These features can lead to elect SERS, among the spectroscopic tools currently at disposal of scientists, as a technique of choice for the identification of anthocyanidins, since: (1) anthocyanidins structurally present the same benzopyrylium moiety and differentiate only for the substitution pattern on their phenyl ring, (2) different species are present in aqueous solution depending on the pH. It will be demonstrated that, while resonance Raman spectra of anthocyanidins are very similar to one another, SER spectra show greater differences, leading to a further step in the identification of such important compounds in diluted solutions by means of vibrational spectroscopy. Moreover, the dependence on the pH of the six most common anthocyanidins, i.e. cyanidin, delphinidin, pelargonidin, peonidin, malvidin and petunidin, is studied. To the best of the authors’ knowledge, a complete SERS study of such important molecules is reported in the present work for the first time.

Phosphoryl transfer from α-d-glucose 1-phosphate catalyzed by Escherichia coli sugar-phosphate phosphatases of two protein superfamily types.

The Cori ester α-d-glucose 1-phosphate (αGlc 1-P) is a high-energy intermediate of cellular carbohydrate metabolism. Its glycosidic phosphomonoester moiety primes αGlc 1-P for flexible exploitation in glucosyl and phosphoryl transfer reactions. Two structurally and mechanistically distinct sugar-phosphate phosphatases from Escherichia coli were characterized in this study for utilization of αGlc 1-P as a phosphoryl donor substrate. The agp gene encodes a periplasmic αGlc 1-P phosphatase (Agp) belonging to the histidine acid phosphatase family. Had13 is from the haloacid dehydrogenase-like phosphatase family. Cytoplasmic expression of Agp (in E. coli Origami B) gave a functional enzyme preparation (kcat for phosphoryl transfer from αGlc 1-P to water, 40 s(-1)) that was shown by mass spectrometry to exhibit no free cysteines and the native intramolecular disulfide bond between Cys(189) and Cys(195). Enzymatic phosphoryl transfer from αGlc 1-P to water in H2 (18)O solvent proceeded with complete (18)O label incorporation into the phosphate released, consistent with catalytic reaction through O-1-P, but not C-1-O, bond cleavage. Hydrolase activity of both enzymes was not restricted to a glycosidic phosphomonoester substrate, and d-glucose 6-phosphate was converted with a kcat similar to that of αGlc 1-P. By examining phosphoryl transfer from αGlc 1-P to an acceptor substrate other than water (d-fructose or d-glucose), we discovered that Agp exhibited pronounced synthetic activity, unlike Had13, which utilized αGlc 1-P mainly for phosphoryl transfer to water. By applying d-fructose in 10-fold molar excess over αGlc 1-P (20 mM), enzymatic conversion furnished d-fructose 1-phosphate as the main product in a 55% overall yield. Agp is a promising biocatalyst for use in transphosphorylation from αGlc 1-P.

Cholesteryl and diosgenyl glycosteroids: synthesis and characterization of new smectic liquid crystals

While present in large numbers in nature, studies on the physical chemical aspects of glycosteroids are quite rare and focused on cholesterol, and all compounds studied thus far have shown liquid crystalline properties in a narrow temperature range. New glycosteroids composed by cholesterol or diosgenin and different glycosidic moieties have been synthesized here in order to analyze the influence of the structure on the formation of mesophases. These compounds have been studied by crossed polarized optical microscopy. These studies have revealed that these new glycosteroids form Smetic A liquid crystals in a broad temperature range.

HPLC-HRMS/MS analysis of saponins and phytoecdysteroids from a Silene colorata hydroglycerinic extract

The genus Silene (family of Caryophyllaceae) is known to contain triterpenoid saponins [1] and phytoecdysteroids [2]. Silene colorata is distributed over the Mediterranean basin, but little is known concerning the phytochemistry of the species. The aim of this work was the development of an analytical method for the rapid characterization of saponins and phytoecdysteroids in a commercially available S. colorata hydroglycerinic extract. After sample preparation on Solid Phase Extraction cartridges, the extract was analyzed by HPLC-HR-MS/MS on an LTQ-Orbitrap platform. The chromatographic and spectrometric conditions applied for the analysis were optimized for the two compound classes. The analysis revealed the presence of three major groups of phytochemicals in the extract, namely di- and tri-glycosylated flavonoids, phytoecdysteroids, and saponins. Among phytoecdysteroids, 20-hydroxyecdysone and a sileneoside derivative (Figure) were the major constituents, while five more major compounds that belong to this category were identified. Saponins detected were molecules of high molecular weights (> 950 and up to 1630) and MS/MS experiments revealed the presence of three to six sugar units on the molecules. In some cases, the glycosidic moiety was esterified with a cinnamic acid derivative, e.g. saponin in the Figure. These structural features of the saponin corresponded with those for saponins in other Silene species [3]. The aglycons of the saponins were characterized with the aid of MS3 experiments. In all cases the triterpenic aglycon gave a pseudomolecular ion of m/z 485.3405, corresponding to a molecular formula of C30H47O5, with 7.5 RDBeq. The molecular formula corresponded to that of quillaic or gypsogenic acids, that have been both reported as characteristic triterpenoidal aglycones in other species [1].

Target-specific imaging of transmembrane receptors using quinonyl glycosides functionalized quantum dots.

Here, we describe a novel "switch-on" biosensor based on quinonyl glycosides functionalized quantum dots (QDs) for the specific targeting and imaging of transmembrane glycoprotein receptors on the surface of cancer cells. The design of the quinonyl glycosides lies in that the quinone moiety serves as a quencher of QDs and the glycoside moiety as a biospecific ligand for targeting a receptor. We observed that the quenched photoluminescence of the quinone glycosides functionalized QDs could be significantly recovered by a specific lectin that selectively binds to the glycosides clustering the QDs but was not affected by a panel of nonspecific lectins. Moreover, we determined that quinonyl galactoside functionalized QDs could optically image the asialoglycoprotein receptors of a hepatoma cell line in a target-specific manner. This system might provide new insights into the fabrication of photoluminogenic biosensors for the analysis of the universal ligand-receptor recognitions in nature.

STUDY OF ANTIADENOVIRALACTIVITY OF NEW FLUORINECONTAINING HETEROCYCLIC COMPOUNDS

Purpose: To study the cytotoxic action and antiviral activity of new fluorinated nucleoside compounds on human adenovirus model and the role of individual molecular fragments in their activity. Methods. Cytotoxic activity of the compounds was determined by the MTT method. Antiviral activity of substances was studied using cyto-morphological method with acridine orange dye. The virus-specific intranuclear inclusions were determined. Results. In present study the cytotoxicity of 2N-substituted-4-tosyl-5-polyfluoroalkyl-1,2,3-triazoles for Hep-2 and Hela cell lines were studied. The dependence of cytotoxicity on the compounds in the structure of substituents in the aromatic rings of the molecules was defined. Combination of residues of 3-hlorotetrahydropyran, 3-chloro-dihydrofuran tetrahydrofuran in a glycosidic moiety with trifluoromethyl or perfluoropropyl in a triazole ring were more toxic. Antiviral activity of 3 compounds (G7, G8 and G10) against HAdV5 in vitro was revealed. The effective antiviral concentrations (EC50) for them were 16, 70 and 186 mg/ml, and the selectivity indexes were equal to 91, 8 and 4, respectively. Conclusions. High rates of antiviral activity and low cytotoxicity of G8 compound make it perspective for development antiviral agents.

Can Amphipathic Helices Influence the CNS Antinociceptive Activity of Glycopeptides Related to β-Endorphin?

Glycosylated β-endorphin analogues of various amphipathicity were studied in vitro and in vivo in mice. Opioid binding affinities of the O-linked glycopeptides (mono- or disaccharides) and unglycosylated peptide controls were measured in human receptors expressed in CHO cells. All were pan-agonists, binding to μ-, δ-, or κ-opioid receptors in the low nanomolar range (2.2–35 nM Ki’s). The glycoside moiety was required for intravenous (i.v.) but not for intracerebroventricular (i.c.v.) activity. Circular dichroism and NMR indicated the degree of helicity in H2O, aqueous trifluoroethanol, or micelles. Glycosylation was essential for activity after i.v. administration. It was possible to manipulate the degree of helicity by the alteration of only two amino acid residues in the helical address region of the β-endorphin analogues without destroying μ-, δ-, or κ-agonism, but the antinociceptive activity after i.v. administration could not be directly correlated to the degree of helicity in micelles.

Anti-complementary constituents of Houttuynia cordata and their targets in complement activation cascade

Activity-guided fractionation for complement inhibitors led to the isolation of 23 known compounds from Houttuynia cordata Thunb. Seven flavonoids, two alkaloids, one coumarin and two phenols showed anti-complementary activity. Preliminary inhibitory mechanism of four flavonoids, including quercitrin, afzelin, isoquercitrin and quercetin in the complement activation cascade were examined for the first time. The results indicated that the target components of flavonols are different from those of flavonosides, and the glycoside moieties may be necessary to block C3 and C4 components.