Modification and inhibition of vancomycin group antibiotics by formaldehyde and acetaldehyde

It is shown that several vancomycin group antibiotics (vancomycin, eremomycin, and avoparcin) undergo spontaneous chemical modifications when kept at room temperature at neutral pH in aqueous solutions containing traces of formaldehyde or acetaldehyde. This chemical modification predominantly results in a mass increase of 12 Da in the reaction with formaldehyde and 26 Da in the case of acetaldehyde. By using tandem mass spectrometry the modification can unambiguously be identified as originating from the formation of a ring-closed 4-imidazolidinone moiety at the N-terminus of the glycopeptide antibiotics, that is, near the receptor binding pocket of the glycopeptide antibiotics. Bioaffinity mass spectrometry shows that this ring-closure results in a dramatically decreased affinity for the peptidoglycan-mimicking D-alanyl-D-alanine receptor. Additionally, in vitro inhibition measurements on two different strains of bacteria have revealed that the modified antibiotics display reduced antibacterial activity. The ring-closure is also shown to have a dissociative effect on the dimerization of the vancomycin-analogue eremomycin. The spontaneous reaction of vancomycin with formaldehyde or acetaldehyde may have implications not only for the clinical use of this class of antibiotics, but also for the effectiveness of these antibiotics when they are used in chiral separation chromatography or capillary electrophoresis.     

Glycopeptide antibiotics and development of inhibitors to overcome vancomycin resistance

This review outlines the history of vancomycin and other key glycopeptide antibiotics and the molecular basis of vancomycin resistance, and focuses on the development of synthetic inhibitors of vancomycin-resistant enzymes VanR, S, A, H and X to overcome resistance. The literature up to July 2001 is reviewed and 119 references are cited.     

Characterisation of a hydroxymandelate oxidase involved in the biosynthesis of two unusual amino acids occurring in the vancomycin group of antibiotics

ORF22 from the chloroeremomycin gene cluster has been cloned, expressed and characterised as a hydroxymandelate oxidase (HmO) that is involved in the formation of both (S)-4-hydroxyphenylglycine and (S)-3,5-dihydroxyphenylglycine.     

Indirect spectrophotometric determination of gentamicin and vancomycin antibiotics based on their oxidation by potassium permanganate

Four simple, accurate, sensitive and economical procedures (A–D) for the estimation of gentamicin sulphate and vancomycin hydrochloride, both in pure form and in pharmaceutical formulations have been developed. The methods are based on the oxidation of the studied drugs by a known excess of potassium permanganate in sulphuric acid medium and subsequent determination of unreacted oxidant by reacting it with amaranth dye (method A), acid orange II (method B), indigocarmine (method C) and methylene blue (method D), in the same acid medium at a suitable λ_max=521, 485, 610 and 664 nm, respectively. The reacted oxidant corresponds to the drug content. Regression analysis of Beer-Lambert plots showed good correlations in the concentration ranges 4–8, 3–8, 4–9 and 5–9 μg ml⁻¹ with gentamicin and 4–8, 1.5–4, 1.5–4 and 3.5–5.5 μg ml⁻¹ with vancomycin for methods A, B, C, and D, respectively. The molar absorptivity, sandell sensitivity, detection and quantification limits were calculated. The stoichiometric ratios for the cited drugs were studied. The optimum reaction conditions and other analytical parameters were evaluated. The influence of the substance commonly employed as excipients with these drugs were studied. The proposed methods were applied to the determination of these drugs in pharmaceutical formulations. The results have demonstrated that the methods are equally accurate and reproducible as the official methods.     

Selective formation of covalent protein heterodimers with an unnatural amino acid

We report a strategy for the generation of heterodimeric protein conjugates using an unnatural amino acid with orthogonal reactivity. This paper addresses the challenges of site-specificity and homogeneity with respect to the synthesis of bivalent proteins and antibody-drug conjugates. There are numerous antibody-drug conjugates in preclinical and clinical development, yet these are based either on nonspecific lysine coupling chemistry or on disulfide modification made difficult by the large number of cysteines in antibodies. Here, we describe a recombinant approach that can be used to rapidly generate a variety of constructs with defined conjugation sites. Moreover, this methodology results in homogeneous antibody conjugates whose biological, physical, and pharmacological properties can be quantitatively assessed and subsequently optimized. As proof of concept, we have generated anti-Her2 Fab-Saporin conjugates that demonstrate excellent potency in vitro.     

Biosynthesis of the vancomycin group of antibiotics: characterisation of a type III polyketide synthase in the pathway to (S)-3,5-dihydroxyphenylglycine

3,5-dihydroxyphenylacetate, a precursor for the non-proteinogenic amino acid 3,5-dihydroxyphenylglycine occurring in glycopeptide antibiotics, is determined to be catalysed by a type III polyketide synthase using malonyl-CoA as a starter unit.     

Spontaneous formation of wurzite-CdS/zinc blende-CdTe heterodimers through a partial anion exchange reaction

Ion exchange of ionic semiconductor nanoparticles (NPs) is a facile method for the synthesis of type-II semiconductor heterostructured NPs with staggered alignment of band edges for photoelectric applications. Through consideration of the crystallographic orientation and strain at the heterointerface, well-designed heterostructures can be constructed through ion exchange reactions. Here we report the selective synthesis of anisotropically phase-segregated cadmium sulfide (CdS)/ cadmium telluride (CdTe) heterodimers via a novel anion exchange reaction of CdS NPs with an organic telluride precursor. The wurtzite-CdS/zinc blende-CdTe heterodimers in this study resulted from spontaneous phase segregation induced by the differences in the crystal structures of the two phases, accompanying a centrosymmetry breaking of the spherical CdS NPs. The CdS/CdTe heterodimers exhibited photoinduced spatial charge separation because of their staggered band-edge alignment.     

Experimental and theoretical studies of the structures and interactions of vancomycin antibiotics with cell wall analogues

Surface-induced dissociation (SID) of the singly protonated complex of vancomycin antibiotic with cell wall peptide analogue (N(alpha),N(epsilon)-diacetyl-L-Lys-D-Ala-D-Ala) was studied using a 6 T Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FT-ICR MS) specially configured for SID experiments. The binding energy between the vancomycin and the peptide was obtained from the RRKM modeling of the time- and energy-resolved fragmentation efficiency curves (TFECs) of the precursor ion and its fragments. Molecular dynamics simulations of the vancomycin, peptide, and vancomycin-peptide complex were carried out to explore the low energy conformations. Density functional theory (DFT) calculations of the geometries, proton affinities, and binding energies were performed for several model systems including vancomycin (V), vancomycin aglycon (VA), N(alpha),N(epsilon)-diacetyl-L-Lys-D-Ala-D-Ala, and noncovalent complexes of VA with N-acetyl-D-Ala-D-Ala and V with N(alpha),N(epsilon)-diacetyl-L-Lys-D-Ala-D-Ala. Comparison between the experimental and computational results suggests that the most probable structure of the complex observed in our experiments corresponds to the neutral peptide bound to the vancomycin protonated at the primary amine of the disaccharide group. The experimental binding energy of 30.9 +/- 1.8 kcal/mol is in good agreement with the binding energy of 36.3-42.0 kcal/mol calculated for the model system representing the preferred structure of the complex.     

Complex formation of chitosan with amikacin and gentamicin antibiotics

Complex formation of chitosan with Amikacin and Gentamicin antibiotics was studied. The optimal conditions for preparing the complexes were found.     

Carbamino group formation with peptides and proteins studied by mass spectrometry

At high pH and in the presence of dissolved CO₂, the N-terminus and ε-amino groups of amino acids, peptides, and proteins can form carbamino adducts with CO₂, R-NH₂ + CO₂ ↔ R-NHCOO⁻ + H⁺. We report the first study of carbamino group formation by electrospray ionization (ESI) mass spectrometry (MS). Angiotensin II, bradykinin, substance P, and insulin have been studied. A careful optimization of the instrumental parameters was necessary to allow the transfer of the fragile adducts into vacuum for mass analysis. Particularly, dissociation of the adducts in the ion sampling process and pH changes in ESI must be minimized. With these precautions, levels of carbamino group formation of angiotensin II and bradykinin determined from mass spectra agree with those expected to be in solution, calculated from literature equilibrium constants. Thus, ESI MS can quantitatively measure ratios of carbamino adduct to total peptide concentration in solution. Values of equilibrium constants for carbamino group formation with substance P (pK_c = 4.77 ± 0.18) and insulin (pK_c = 4.99 ± 0.05) are reported for the first time.     

The synthesis of tethered ligand dimers for PPARgamma-RXR protein heterodimers

Heterodimeric compounds based on tethering the PPARgamma agonist Rosiglitazone to the RXR ligand Targretin have been prepared.     

Complex formation of macrotetrolide carrier antibiotics with cations studied by microcalorimetry and vapour pressure osmometry

Complex formation parameters of macrotetrolide antibiotics with alkali and alkaline earth metal cations are given. The stability constants for the complexes in methanol and ethanol at 30°, as determined by vapour pressure osmometry, and ΔH⁰, ΔG⁰, and ΔS⁰ for some interactions in methanol and ethanol at 25°, measured by microcalorimetry, are compared and discussed.     

The effect of phosphate group replacement by sulfate groups on the phase formation in the synthesis of hydroxyapatite

Sulfate-substituted hydroxyapatite materials with a degree of substitution of up to 20 mol % (Ca₁₀(PO₄)_{(6 – 0.06x)}(SO₄)_0.09x (OH)₂, x = 0, 0.1, 0.5, 1, 5, 10, and 20) were synthesized. For substitutions of 0, 0.1, 0.5, and 1 mol %, a single-phase material with the apatite structure is formed. On further increase in the concentration of SO₄ ^2? groups up to 20 mol %, a second phase, CaSO₄, is formed; the amount of this phase increases for higher degrees of substitution. The unit cell parameters of hydroxyapatite-based materials change slightly upon the replacement of phosphate groups by sulfate groups: the parameter a tends to increase, while c tends to decrease. The introduction of sulfate groups results in decreasing particle size.     

Solid-phase synthesis of disulfide heterodimers of peptides

[structure: see text] The synthesis of the orthogonal disulfide template 1 and its use to synthesize unsymmetrical intermolecular disulfide bond peptides on a solid support are described. Application of template 1 to synthesize bioconjugates of cell permeable moieties based on the disulfide bond is demonstrated.     

Generic nonextractive spectrophotometric method for determination of 4-quinolone antibiotics by formation of ion-pair complexes with beta-naphthol

This paper describes the development of a generic spontaneous nonextractive spectrophotometric method for determination of 13 pharmaceutically important 4-quinolone antibiotics. The method was based on the formation of yellow-colored water-soluble ion-pair complexes between 2% (w/v) beta-naphthol reagent and each of the studied drugs in sulfuric acid medium at room temperature. The formed ion-pair chromogens have maximum absorption peaks in the range of 365-391 nm. The concentrations of the reagents and the experimental conditions affecting the reaction were optimized. Under the optimum conditions, linear relationships with good linear coefficients (0.9987-0.9995) were found between the absorbance and concentration of the investigated drugs in the range of 10-350 microg/mL. The assay limits of detection and quantitation were 1-9.9 and 3.4-32.9 microg/mL, respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 2%. The proposed method was successfully applied to the analysis of the investigated drugs in pure and pharmaceutical dosage forms with good accuracy and precision; the percentages of label claim ranged from 97.8-102.8 +/- 0.35-1.60%. The results obtained by the proposed spectrophotometric method were comparable with those obtained by the official or reported methods. The proposed method is superior to all the previously reported ion-pair formation-based methods in terms of simplicity because it did not involve extraction procedures for the ion-pair complex. Therefore, this method might be recommended for routine use in quality control laboratories for analysis of the investigated 4-quinolone antibiotics in their pure forms, as well as in pharmaceutical dosage forms.     

Nitro group participation in the formation of heterocycles

Benzoyl bromide (2-nitrophenyl) hydrazone (2) was treated with sodium ethoxide and ethyl cyanoacetate and two unexpected products were obtained. These products were ultimately shown to be 6-bromo-3-phenyl-1,2,4-benzotriazine (32) and 5-bromo-2-phenylbenzoxazole (38), by comparison with authentic samples which were synthesized. A mechanism is presented for the formation of these two heterocyclic systems (32 and 38) from 2.     

Hydroxyl group formation in the dehydration of scolecite

Conclusions The formation of OH groups occurs at pumping temperature 503 K and is complete at 623 K.Translated from Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, No. 3, pp. 728–729, March, 1989.