Formation Of Glycosylamines Research Articles

Insights into the Progression of Labile Hb A1c to Stable Hb A1c via a Mechanistic Assessment of 2,3-Bisphosphoglycerate Facilitation of the Slow Nonenzymatic Glycation Process

Nonenzymatic glycation (NEG) of human hemoglobin (Hb A) consists of initial non covalent, reversible steps involving glucose and amino acid residues, which may also involve effector reagent(s) in the formation of labile Hb A1c (the conjugate acid of the Schiff base). Labile Hb A1c can then undergo slow, largely irreversible, formation of stable Hb A1c (the Amadori product). Stable Hb A1c is measured to assess diabetic progression after labile Hb A1c removal. This study aimed to increase the understanding of the distinctions between labile and stable Hb A1c from a mechanistic perspective in the presence of 2,3-bisphosphoglycerate (2,3-BPG). 2,3-Bisphosphoglycerate is an effector reagent that reversibly binds in the Hb A1c pocket and modestly enhances overall NEG rate. The deprotonation of C2 on labile Hb A1c in the formation of the Amadori product was previously proposed to be rate-limiting. Computational chemistry was used here to identify the mechanism(s) by which 2,3-BPG facilitates the deprotonation of C2 on labile Hb A1c. 2,3-Bisphosphoglycerate is capable of abstracting protons on C2 and the α-nitrogen of labile Hb A1c and can also deprotonate water and/or amino acid residues, therefore preparing these secondary reagents to deprotonate labile Hb A1c. Parallel reactions not leading to an Amadori product were found that include formation of the neutral Schiff base, dissociation of glucose from the protein, and cyclic glycosylamine formation. These heretofore under appreciated parallel reactions may help explain both the selective removal of labile from stable Hb A1c and the slow rate of NEG.

A concise approach to the synthesis of the unique N-mannosyl d-β-hydroxyenduracididine moiety in the mannopeptimycin series of natural products.

The asymmetric synthesis of the orthogonally protected N-mannosyl d-β-hydroxyenduracididine (N-Man-d-βhEnd) is described, starting from enantiopure silylated (S)-serinol. The key steps are: (i) glycosylamine formation between protected serinol and a benzylated d-mannose; (ii) guanidinylation; and (iii) cyclic guanidine formation. This synthesis constitutes a breakthrough in our studies towards a total synthesis of mannopeptimycin and should also allow for other studies in the field of mannopeptimycin research, including the synthesis of derivatives.

The Maillard reaction of bisoprolol fumarate with various reducing carbohydrates

HPLC analysis of drug products containing bisoprolol fumarate and lactose revealed the presence of N-formylbisoprolol, which is a final product of the Maillard reaction. Formulations containing secondary amines and reducing carbohydrates are prone to the condensation of amine and carbonyl functional groups and formation of glycosylamines in pharmaceutically relevant conditions. Further rearrangement occurs in the presence of a nucleophile and leads to the formation of 1-deoxy-1-amino-2-ketose also known as the Amadori Rearrangement Product (ARP). The influence of water content, carbohydrate, and lubricant types on the reaction rate was tested. The reaction progress was monitored by HPLC and UV–Vis spectrophotometry. The structures of intermediates were confirmed by the LC/MS2 analysis. N-formylbisoprolol – the final reaction product – was synthesised and characterised by LC/MS2, H1 and C13 NMR.

18F‐Fluorodeoxyglycosylamines: Maillard reaction of 18F‐fluorodeoxyglucose with biological amines†

The Maillard reaction of sugars and amines resulting in the formation of glycosylamines and Amadori products is of biological significance, for drug delivery, role in central nervous system, and other potential applications. We have examined the interaction of (18) F-fluorodeoxyglucose ((18) F-FDG) with biological amines to study the formation of (18) F-fluorodeoxyglycosylamines ((18) F-FDGly). Respective amines N-allyl-2-aminomethylpyrrolidine (NAP) and 2-(4'-aminophenyl)-6-hydroxybenzothiazole (PIB precursor) were mixed with FDG to provide glycosylamines, FDGNAP and FDGBTA. Radiosynthesis using (18) F-FDG (2-5 mCi) was carried out to provide (18) F-FDGNAP and (18) F-FDGBTA. Binding of FDGBTA and (18) F-FDGBTA was evaluated in human brain sections of Alzheimer's disease (AD) patients and control subjects using autoradiography. Both FDGNAP and FDGBTA were isolated as stable products. Kinetics of (18) F-FDGNAP reaction indicated a significant product at 4 h (63% radiochemical yield). (18) F-FDGBTA was prepared in 57% yield. Preliminary studies of FDGBTA showed displacement of (3) H-PIB (reduced by 80%), and (18) F-FDGBTA indicated selective binding to Aβ-amyloid plaques present in postmortem AD human brain, with a gray matter ratio of 3 between the AD patients and control subjects. We have demonstrated that (18) F-FDG couples with amines under mild conditions to form (18) F-FDGly in a manner similar to click chemistry. Although these amine derivatives are stable in vitro, stability in vivo and selective binding is under investigation.

ChemInform Abstract: Synthesis of 1,2- and 1,3-N-Linked Disaccharides of 5-Thio-α-D-mannopyranose as Potential Inhibitors of the Processing Mannosidase Class I and Mannosidase II Enzymes.

The syntheses of S/N acetal heteroatom analogues of 1,2- and 1,3-linked mannopyranose disaccharides are described. The compounds are analogues of the Man-α-(1→2)-Man and Man-α-(1→3)-Man disaccharide components of oligosaccharides found in N-glycoproteins that are cleaved by trimming mannosidases during glycoprotein processing. Glycosylamine formation, without the necessity of hydroxyl group protection, proceeded through acid-catalyzed condensation reactions of 5-thio-d-mannose with either methyl 2-amino-2-deoxy- or 3-amino-3-deoxy-α-d-mannopyranoside to give methyl 2-amino-2-deoxy-2-N-(5-thio-α/β-d-mannopyranosyl)-α-d-mannopyranoside (2) or methyl 3-amino-3-deoxy-3-N-(5-thio-α/β-d-mannopyranosyl)-α-d-mannopyranoside (3), respectively. The superiority of mercuric chloride over acetic acid as a catalyst for this reaction is reported. Acetylation of the anomeric mixtures gave the heptaacetates from which the major β‘-isomers could be separated by chromatography and/or crystallization.

Glycosylation of aromatic amines II: Kinetics and mechanisms of the hydrolytic reaction between kynurenine and glucose

The kinetics of the weakly basic aromatic amine, kynurenine, with glucose were studied as model reactants aimed at mechanistic understanding of pharmaceutically relevant amine-aldehyde reactions. The reaction kinetics of the forward and reverse processes (glycosylamine formation and hydrolysis) were studied under first-order conditions in aqueous solutions at 40°C in the pH range 1–6.5 in the presence of various buffers. The α-and β-glycosylamines were reversibly formed via an acyclic imine that was not present in detectable quantities. Rate-limiting formation of the imine was complex and involved the addition of the amine and aldehyde to form the carbinolamine followed by the acid-catalyzed dehydration to the imine. The pH-rate profile was characterized by three kinetically distinguishable processes. At lower pH values, the profile was consistent with specific acid-catalyzed rate-determining addition of amine and aldehyde. In the pH range of 4–6 a downward bend was attributable to the change in rate determining step from addition to dehydration. In the pH region of 2–3 the rate law was described by specific acid catalysis and solvolysis of the zwitterionic form of kynurenine. Nonlinear buffer effects and Brönsted plots were shown to be consistent with this interpretation of the pH-rate profile. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4616–4628, 2009

Glycosylation of Aromatic Amines I: Characterization of Reaction Products and Kinetic Scheme

The reactions of aliphatic and aromatic amines with reducing sugars are important in both drug stability and synthesis. The formation of glycosylamines in solution, the first step in the Maillard reaction, does not typically cause browning but results in decreased potency and is hence significant from the aspect of drug instability. The purpose of this research was to present (1) unreported ionic equilibria of model reactant (kynurenine), (2) the analytical methods used to characterize and measure reaction products, (3) the kinetic scheme used to measure reaction rates and (4) relevant properties of various reducing sugars that impact the reaction rate in solution. The methods used to identify the reversible formation of two products from the reaction of kynurenine and monosaccharides included LC mass spectrometry, UV spectroscopy, and 1-D and 2-D (1)H-(1)H COSY NMR spectroscopy. Kinetics was studied using a stability-indicating HPLC method. The results indicated the formation of alpha and beta glycosylamines by a pseudo first-order reversible reaction scheme in the pH range of 1-6. The forward reaction was a function of initial glucose concentration but not the reverse reaction. It was concluded that the reaction kinetics and equilibrium concentrations of the glycosylamines were pH-dependent and also a function of the acyclic content of the reacting glucose isomer.

Synthesis of 1,2- and 1,3-N-Linked Disaccharides of 5-Thio-α-d-mannopyranose as Potential Inhibitors of the Processing Mannosidase Class I and Mannosidase II Enzymes

The syntheses of S/N acetal heteroatom analogues of 1,2- and 1,3-linked mannopyranose disaccharides are described. The compounds are analogues of the Man-alpha-(1-->2)-Man and Man-alpha-(1-->3)-Man disaccharide components of oligosaccharides found in N-glycoproteins that are cleaved by trimming mannosidases during glycoprotein processing. Glycosylamine formation, without the necessity of hydroxyl group protection, proceeded through acid-catalyzed condensation reactions of 5-thio-D-mannose with either methyl 2-amino-2-deoxy- or 3-amino-3-deoxy-alpha-D-mannopyranoside to give methyl 2-amino-2-deoxy-2-N-(5-thio-alpha/beta-D-mannopyranosyl)-alpha-D-mannopyranoside (2) or methyl 3-amino-3-deoxy-3-N-(5-thio-alpha/beta-D-mannopyranosyl)-alpha-D-mannopyranoside (3), respectively. The superiority of mercuric chloride over acetic acid as a catalyst for this reaction is reported. Acetylation of the anomeric mixtures gave the heptaacetates from which the major beta'-isomers could be separated by chromatography and/or crystallization.

Effect of storage temperature and time on fiber content of fresh and ensiled alfalfa.

To determine effects of storage temperature and time on fiber content, fresh alfalfa (late vegetative) was hand-clipped from three locations (blocks) and stored fresh or ensiled at -20 degrees C or +60 degrees C for 0, 24, 48, 96, 168, or 336 h. The experimental design was a randomized complete block with a 2 x 2 factorial arrangement of treatments. Main effects were method of forage conservation (fresh or ensiled) and temperature of storage (-20 degrees C or + 60 degrees C). Length of storage (0, 24, 48, 96, 168, or 336 h) was in the subplot. Conservation method x storage temperature x time interactions were detected (P < .05) for NDF, IVDMD, ADIN, and N content of NDF. Large increases in alfalfa NDF were observed up to 168 h of storage at 60 degrees C, whereas when fresh alfalfa was stored at -20 degrees C, small increases were observed up to 48 h but a large increase was observed after 168 h. Ensiled alfalfa stored at -20 degrees C had no change in NDF content. Contributors to increased NDF seemed due to changes in protein solubility and glycosylamine formation when fresh forage was stored at 60 degrees C, and changes in protein solubility, glycosylamine formation, and Maillard product formation when ensiled forage was stored at 60 degrees C. At -20 degrees C, formation of adducts with arabinose and glucose from nonstructural carbohydrates as well as polymerization and fragmentation of cell wall monosaccharides probably contributed to altered NDF content of alfalfa.

Linkage Analysis of Chromophore-Labeled Disaccharides and Linear Oligosaccharides by Negative Ion Fast Atom Bombardment Ionization and Collisional-Induced Dissociation with B/E Scanning

Negative ion fast atom bombardment ionization in combination with collisional-induced dissociation mass spectrometry differentiates the linkage position(s) of chromophore-labeled di-. and oligosaccharides. The formation of glycosylamines rather than the more popular reductive amination has been used to label the reducing end of sugars with ultraviolet or fluorescence tags. Two types of fragment ions were detected, one with the charge carried on the chromophore end and the other with the charge carried on the nonreducing terminus. These ions are essential to the determination of interglycosidic linkage and they are produced from the ring cleavage of the reducing end monosaccharide. Additionally, the anomeric configuration of the 1-4-linked residue could be assigned according to the relative abundance of the fragment ions.

Formation of the N,N'-dialkylpyrazine cation radical from glyoxal dialkylimine produced on reaction of a sugar with an amine or amino acid.

The formation of an intermediate product, which could easily give a radical product, in an early stage of the Maillard reaction was confirmed commonly occur in various sugar-amino compound systems, by detection of the N, N'-dialkylpyrazine cation radical generated on the addition of ascorbic acid (AsA) to the reaction mixtures. This intermediate was produced immediately after to glycosylamine formation, prior to Amadori rearrangement, and completely parallel to the formation of glyoxal dialkylimine, which was identified by TLC as a main component of the extract. Authentic glyoxal dialkylimine was shown to produce an identical radical on treatment with both reducing agents and acids instead of AsA. It was thus demonstrated that the intermediate is glyoxaldialkylimine and that acid hydrolysis followed by reduction is required for production of the free radical.

Formation of novel nucleosides from free base and sugar phosphate: Aqueous reaction of 2-aminopyrimidine and ribose-5-phosphate

The facile formation of glycosylamines suggests that a base liberated by depurination might react at the free C 1 position of the sugar phosphate from which it had been hydrolyzed, effectively repurinating the site. Model experiments testing this hypothesis demonstrate that such a reaction does take place. The primary product of a reaction between 2-aminopyrimidine (a model for guanine) and ribose-5-phosphate is characterized by enzymatic and chemical degradation, and UV spectra. It is shown to be a novel nucleoside with the base attached via its exocyclic amino group to the C 1 of the ribose-5-phosphate.

Thermolysis of derivatives of amino sugars

Amino groups influence the course of the pyrolysis of carbohydrate derivatives and result in substantial dehydration and charring. N-Phenylglycosylamines and their acetates rearrange with almost quantitative loss of water or acetic acid and retention of the arylamino groups. The sharp dehydration reaction contrasts with the behaviour of the corresponding phenyl glycosides on thermolysis, which results in cleavage of the phenolic groups and subsequent breakdown of the glycosyl moiety. Thermal analysis and chemical data indicate that charring of other derivatives of amino sugars is due to intermediate formation of glycosylamines from thermal reaction of the amino group with glycosidic centres.

The Maillard Reaction

This chapter discusses the Maillard reaction. Results of the many investigations into the mechanism of the Maillard reaction support one of two main theories. The first assumes the formation of glycosylamines that undergo the Amadori (or, for ketoses, the Heyns) rearrangement. The 1-amino-l-deoxyketose derivative (or 2-amino-2-de- oxyaldose derivative) formed may be dehydrated and cyclized to form furan derivatives, or it may enolize. In either case, intermediates that are readily transformed into brown compounds are formed. A third possibility is for the deoxy sugar derivative to react with more amino acid to form colored products. The many workers who have supported this mechanism found also that optimum conditions for occurrence of the Maillard reaction are (1) fairly low water content, (2) a p H of 7 to 10, and (3) a high temperature. Nevertheless, some reaction occurs under conditions far removed from these, but in the absence of moisture there is no reaction. The formation of an acyclic Schiff base as an initial step is not very likely, since replacement of the aldose by salicylaldehyde caused only a very small loss of amino groups. The second theory of the mechanism of the browning reaction is of recent origin and maintains that the browning reaction and the Maillard reaction are separate and distinct. Browning, according to this school of thought, is due to the effect of p H on the sugar and can occur over a wide range of p H, whereas the Maillard reaction proceeds only in alkaline media.