Elucidation of chiral recognition processes of macrocyclic antibiotic vancomycin

A theoretical investigation was carried out on the retention and separation of enantiomeric molecules including nonsteroidal anti-inflammatory drugs, anti-neoplastic compounds and N-derivatized amino acids by capillary electrophoresis using macrocyclic antibiotics, a new class of chiral selectors, as stationary phase. Firstly docking methods were used to study the enantiorecognition in chiral electrophoresis. The molecular dynamics simulations of the two diastereoisomer complexes were then performed in order to understand how these antibiotics recognize the enantiomers. Another approach was applied in this study to establish a quantitative structure-enantioselectivity relationship (QSER) model, able to describe the resolution of a series of chiral compounds in capillary electrophoresis using vancomycin as the resolving agent.     

Enantiomer separation by capillary electrophoresis using DEAE-dextran and aminoglycosidic antibiotics

Summary Diethylaminoethyl (DEAE) dextran hydrochloride and three kinds of aminoglycosidic antibiotics; fradiomycin sulfate, kanamycin sulfate and streptomycin sulfate, were employed as chiral selectors in capillary electrophoresis, enantiomer separation. These selectors are cationic or basic because of amino functionality and therefore used for enantiomer separation of acidic compounds. To avoid adsorption of the basic or cationic selectors on the capillary inner surface, a coated capillary was employed. Among those tested, enantiomers of binaphthyl compounds and synthetic intermediates of diltiazem analogues were separated. Methanol addition was effective for the improvement of peak shape and resolution.     

Evaluation of the enantioseparation capability of the novel chiral selector clindamycin phosphate towards basic drugs by micellar electrokinetic chromatography

To date, a series of chiral selectors have been utilized successfully in capillary electrophoresis (CE). Among these various chiral selectors, macrocyclic antibiotics have been demonstrated to represent powerful enantioselectivity towards many chiral compounds. Differing from macrocyclic antibiotics, the use of lincosamide antibiotics as chiral selectors has not been reported previously. In our recent work, clindamycin phosphate belonging to the group of lincosamides has been first used as a chiral selector in capillary zone electrophoresis (CZE). In this paper, a micellar electrokinetic chromatography (MEKC) method has been developed for the evaluation of enantioseparation capability of this novel chiral selector towards several racemic basic drugs. As observed during the course of this work, clindamycin phosphate allowed excellent separation of the enantiomers of nefopam, citalopram, tryptophan, chlorphenamine, propranolol and metoprolol, as well as partial enantioresolution of tryptophan methyl ester and cetirizine. In this MEKC chiral separation system, different types of anionic surfactants, organic additives and background electrolytes were tested, and satisfactory enantioseparations of basic drugs above-mentioned were achieved using sodium dodecyl sulfate (SDS) as the surfactant, isopropanol as the organic additive, and phosphate as the background electrolyte. Furthermore, both migration times and enantioseparation of the analytes were influenced by several experimental parameters such as pH of the BGE, clindamycin phosphate and SDS concentrations, phosphate and isopropanol concentrations, and applied voltage. Consequently, the effects of these factors on enantioseparations of the studied basic drugs were systematically investigated in order to evaluate the stereoselectivity of clindamycin phosphate in MEKC.     

Evaluation of balhimycin as a chiral selector for enantioresolution by capillary electrophoresis

The glycopeptide antibiotic balhimycin and its haloanalogue bromobalhimycin were evaluated as chiral selectors for enantioresolution by capillary electrophoresis. In order (i) to eliminate the adsorption of the glycopeptide antibiotics on the capillary wall, (ii) to shorten the separation time and (iii) to improve the detection sensitivity, a combined approach of the dynamic surface coating technique, the co-electroosmotic flow electrophoresis technique and the partial filling technique was employed for the enantioresolution of 16 acidic racemates. The effect of experimental parameters (plug length of the partial filling solution containing the chiral selector, selector concentration and buffer pH) on enantiorecognition was investigated. Furthermore, the enantiorecognition ability imparted by balhimycin, bromobalhimycin and vancomycin were compared. For most tested compounds, the highest enantiorecognition was obtained with balhimycin as chiral selector. Only in the case of the enantioresolution of tiaprofenic acid, vancomycin showed a superior enantiorecognition.     

Separation of binaphthyl enantiomers by capillary zone electrophoresis and electrokinetic chromatography

The capillary electrophoretic (CE) separation of the enantiomers of three binaphthyl compounds is investigated. Several CE modes such as cyclodextrin (CD) modified capillary zone electrophoresis (CZE) (CD-CZE), micellar electrokinetic chromatography (MEKC), cyclodextrin electrokinetic chromatography (CD-EKC), etc. are employed for the simultaneous enantiomer separation of the three solutes. The successful separation was achieved by combining two modes, in other words by using more than two chiral selectors. A development of the CE enantiomer separation is demonstrated for the binaphthyl compounds. The enantioselectivity of binaphthyl compounds is alo briefly discussed.     

A further study on the chiral separation power of a soluble neutral β-cyclodextrin polymer

Enantiomeric separation of some basic compounds, namely selegiline, amphetamine, and clenbuterol, was studied by capillary electrophoresis using an uncharged β-cyclodextrin polymer added to the background electrolyte at pH 2.5. Both complexation and resolution were influenced by the concentration of the chiral polymer confirming our previous results obtained in our earlier work for a wide number of pharmaceutical compounds. In this further study, we examined the influence of different organic additives to the background electrolyte on the enantioselectivity of the chiral selector, also using an extended number of analytes. In most cases, the use of an organic additive resulted in a decrease of resolution. However opposite to that, in some cases, e.g. ephedrine, the organic solvent proved to be essential to achieve enantiomeric resolution. Furthermore the influence of the capillary temperature on the resolution of the analytes was evaluated. Increase of temperature had a deleterious effect on the resolution of the enantiomers. For ephedrine, however, relatively high temperature (50 °C) proved to be advantageous, for the resolution of the optical isomers.     

Enantioseparation of dihydrofurocoumarin derivatives by various separation modes of capillary electrophoresis

Chiral dihydrofurocoumarin compounds are currently the focus of industrial and pharmacological research. These derivatives have been shown to possess many physiological properties that could be medically beneficial. This work proposes four different chiral separation methods using capillary electrophoresis and micellar capillary electrophoresis (MCE). Several different cyclodextrin chiral selectors were examined to evaluate their effectiveness in the enantioseparation of dihydrofurocoumarins. In addition, the effects of the chiral selector concentration, the presence of an organic modifier, run buffer pH, and in two cases, the ratio between the chiral selector and an additional charged pseudophase were investigated. Overall, the best separations for this class of chiral compounds were achieved using sulfated beta-cyclodextrins at low pH in the reversed polarity mode.     

Azithromycin as a new chiral selector in capillary electrophoresis

In capillary electrophoresis (CE), separation of enantiomers of a chiral compound can be achieved through the chiral interactions and/or complex formation between the chiral selector and the enantiomeric analytes on leaving their diastereomeric forms with different stability constants and hence different mobilities. A great number of chiral selectors have been employed in CE and among them macrocyclic antibiotics exhibited excellent enantioselective properties towards a wide number of racemic compounds. The use of azithromycin (AZM) as a chiral selector has not been reported previously. This work reports the use of AZM as a chiral selector for the enantiomeric separations of five chiral drugs and one amino acid (tryptophan) in CE. The enantioseparation is carried out using polar organic mixtures of acetonitrile (ACN), methanol (MeOH), acetic acid and triethylamine as run buffer. The influences of the chiral selector concentration, ACN/MeOH ratio, applied voltage and capillary temperature on enantioseparation are investigated. The results show that AZM is a viable chiral selector in CE for the enantioseparation of the type of chiral drugs investigated.     

Chiral analysis of UV nonabsorbing compounds by capillary electrophoresis using macrocyclic antibiotics: 1. Separation of aspartic and glutamic acid enantiomers

Glycopeptide antibiotics, namely vancomycin or teicoplanin, were evaluated in capillary electrophoresis for the analysis of UV nonabsorbing compounds such as aspartic and glutamic acid enantiomers. Electrophoretic runs were performed in laboratory-made polyacrylamide-coated capillaries using the partial filling-counter current method in order to avoid the presence on the detector path of the absorbing chiral selector. The background electrolyte consisted of an aqueous or aqueous-organic buffer in the pH range of 4.5-6.5 of sorbic acid/histidine and the appropriate concentration of chiral selector. Several experimental parameters such as antibiotic concentration and type, buffer pH, organic modifier, type and concentration of absorbing co-ion (for the indirect UV detection) were studied in order to find the optimum conditions for the chiral resolution of the two underivatized amino acids in their enantiomers. Among the two investigated chiral selectors, vancomycin resulted to be the most useful chiral selector allowing relatively high chiral resolution of the studied compounds even at low concentration. The optimized method (10 mM sorbic acid/histidine, pH 5, and 10 mM of vancomycin) was used for the analysis of real samples such as teeth dentine and beer.     

Enantioselective separations in capillary electrophoresis with dextran sulfate as the chiral selector

Dextran sulfate, a polyanionic polysaccharide, was evaluated as a chiral additive in capillary electrophoresis. Structurally related compounds having a variety of functional groups were utilized to probe the selectivity of the chiral selector. The effects of pH, chiral selector concentration, and chiral selector composition on resolution were also studied. At low pH, the reversed polarity mode was employed to achieve separation of the probe compounds. The electrophoretic results provided insight into the chiral recognition of dextran sulfate in capillary electrophoresis. Several factors, including hydrophobic, steric, and electrostatic interactions, appeared to play a role in the observed enantioseparations.     

亲和毛细管电泳研究生物分子的手性识别*

结合作者研究组近年来对毛细管电泳的系统研究,评述了亲和毛细管电泳方法研究生物分子的手性识别进展。简要介绍了该领域研究的意义、原理,列举了含有蛋白、抗生素、糖及核酸等4种重要生物分子的作用体系及一些应用,并展望了该领域的发展潜力及前景。     

Chiral selectivity of guanosine media in capillary electrophoresis

Gels formed by self-association of monomeric guanosine compounds join numerous other agents such as cyclodextrins, crown ethers, chiral surfactants, antibiotics, proteins, and polysaccharides for chiral separations. Guanosine gels (G-gels) are self-assembled networks of hydrogen-bonded tetrads formed by guanosine nucleotides and their derivatives. The tetrads stack upon themselves to form columnar, helical aggregates that are stabilized by π-π interactions and centrally located cations. Previous work showed the effectiveness of G-gels formed by guanosine-5'-monophophate for separation of the enantiomers of the cationic drug propranolol using capillary electrophoresis. Subsequently, it was found that not all chiral compounds could be resolved into their enantiomers, leading us to investigate in this work the structural features that appear to be correlated to enantiomerically selective interactions of chiral compounds with G-gels. For those compounds (anionic 1,1'-binaphthyl-2,2'-diyl hydrogen phosphate and zwitterionic tryptophan) for which enantiomeric resolution was achieved, the effects of experimental conditions and G-gel composition were examined. For other compounds with no net charge (hydrobenzoin and zwitterionic amino acids and derivatives), the migration times were used as an indicator of the extent of interaction with the G-gel run buffer. It was found that the extent of interaction alone does not determine the chiral selectivity of the G-gel, indicating that the mechanism of chiral separation involves particular structural characteristics of the chiral compounds.     

Chiral separation by capillary electrophoresis with oligosaccharides.

Maltodextrins, i.e., mixtures of linear alpha-(1-4)-linked D-glucose polymers, were found to be effective as chiral electrolyte modifiers to perform direct, rapid separations by capillary electrophoresis of racemic mixtures of 2-arylpropionic acid non-steroidal anti-inflammatory compounds and coumarinic anticoagulant drugs, and also diastereomeric cephalosporin antibiotics. Enantioselectivity seemed to be dependent on an as yet unidentified combination of variables.     

Sensitive Method for Enantioseparation of Rivastigmine with Highly Sulfated Cyclodextrin as Chiral Selector by Capillary Electrophoresis

A sensitive method for enantioseparation of a basic drug rivastigmine and determination of its optical impurityby capillary electrophoresis with highly sulfated β-cyclodextrin(HS-β-CD)as the chiral selector is described.Ingeneral,enantioseparation of basic chiral compounds is carried out in acidic condition(pH 2.5)to prevent analytesfrom adsorption on the capillary wall.However,in the case of rivastigmine,the detection sensitivity was too limitedto determine the optical impurity of S-rivastigmine lower than 1% when buffer pH was 2.5.It was found that thedetection sensitivity was improved 1.6 times just by raising the buffer pH value from 2.5 to 5.8.The poor columnefficiency due to the adsorption of the analytes on the capillary wall was resolved by using dynamical coating of thecapillary wall with the linear polyacrylamide solution.The experirnental parameters such as the concentration ofHS-β-CD,buffer pH and buffer ionic strength were optimized,respectively.The method was validated in terms ofrepeatability,linearity,limit of detection(LOD)and limit of quantitation(LOQ).Using the present method,the op-tical purity of nonracemic rivastigmine with the enantiomeric excess(ee)value of 99.14% was determined.     

Influence of the Buffer Organic Cation on the Chiral Separation of Some Basic Drugs by Capillary Zone Electrophoresis

This paper addresses the influence of small organic cations in the background electrolyte (BGE) on chiral separation by capillary zone electrophoresis (CZE). Seven basic drugs, viz. alprenolol, amphetamine, anisodamine, isoprenaline, pinacidil, synephrine, and verapamil were used as test compounds with hydroxylpropyl-β-cyclodextrin (HP-β-CD) as the chiral selector. Resolution of the chiral test compounds was studied at pH 2.5 in the presence of Tris, ethanolamine, diethanolamine, triethanolamine, diethylamine, and triethylamine, which exist in their cationic forms at this pH value. Among the six cationic additives studied, triethylamine is most effective in enhancing resolution. A tentative mechanism of the effect is proposed.     

A study on the chiral recognition mechanism of enantioseparation of adrenaline and its analogues using capillary electrophoresis

The purpose of this paper was to study the chiral recognition mechanism of enantioseparation of adrenaline and its analogues using capillary electrophoresis. The enantiomeric separation of adrenaline and its analogues has been developed using beta-cyclodextrins as the chiral selectors. All the tested compounds were separated under the same experimental conditions to study the chiral recognition mechanisms, using a low-pH buffer (50 mM Tris buffer at pH 2.5). By means of molecular docking the inclusion course between beta-cyclodextrins and enantiomers was investigated and thus the interaction energy was obtained by molecular mechanics calculations. The results suggest that the difference in interaction energy for the side chain part is most likely responsible for enantiomeric separation.     

Recent developments in chiral capillary electrophoresis and applications of this technique to pharmaceutical and biomedical analysis

This paper provides an overview of the current status of chiral capillary electrophoresis (CE). The emphasis is placed on the application of CE in chiral separation of various racemic compounds. During the last two years about 280 papers, several review articles, and two entire issues, edited by S. Fanali (Electrophoresis 1999, 20, 2577-2798, and H. Nishi and S. Terabe (J. Chromatogr. A 2000, 879, 1-471.) have been devoted to chiral CE. Enantiomeric separations of various compounds, e.g., pharmaceuticals, drug candidates, drugs and related metabolites in biological fluids, amino acids, di- and tri peptides, pesticides and fungicides, have been performed using different chiral selectors. Native and derivatized cyclodextrins continue to be the most widely used chiral selectors. Other chiral selectors such as natural and synthetic chiral micelles, crown ethers, chiral ligands, proteins, oligo- and polysaccharides, and macrocyclic antibiotics have also been applied to chiral CE separations.     

卡那霉素作为手性选择剂的毛细管电泳手性药物分离研究

建立了一种以天然易得的卡那霉素为手性添加剂，用毛细管区带电泳法快速分离市售对乙肝有良好治疗效果的药物联苯双脂衍生物的方法，拓宽了毛细管电泳中手性选择剂的范围，通过实验研究了卡那霉素、甲醇 含量PH值，磷酸盐缓冲体系和硼硝缓冲体系对手性分离的影响，以及三种有机溶剂（甲醇、乙晴、异丙醇）添加剂对手性分离的影响，结果表明，在含有3％卡那霉素，30mol/L，硼砂缓冲体系（PH＝8．0）添加30％异丙醇是最佳的分离条件。     

Separation of enantiomers of <Emphasis Type="Italic">N</Emphasis>-derivatives of amino acids by capillary electrophoresis using macrocyclic antibiotics

The potential for the application of macrocyclic antibiotic eremomycin as a chiral selector for the separation of enantiomers of N-derivatives of amino acids in capillary electrophoresis was estimated. The influence of several factors (the composition and pH of the running electrolyte, the concentration of the chiral selector, capillary geometry, the value of the applied voltage) on the migration of the derivatives of amino acids and enantioselectivity in the presence of eremomycin was studied in order to choose the separation conditions. Using the example of the dansyl derivatives of amino acids the enantioselectivities of vancomycin and eremomycin in capillary electrophoresis were compared.     

Facile routes for the preparation of 3,4‐disubstituted 1,3‐oxazolidines and 1,2,5‐trisubstituted imidazolidin‐4‐ones

Facile, alternative synthetic routes to 6 , (R)‐6 , and (S)‐6 ‐3‐benzyl‐N‐(2,6‐dimethylphenyl)‐1,3‐oxazolidine‐4‐carboxamides ( 6 ), a chiral oxazolidine derivative of tocainide, are reported. The synthetic routes described herein also afforded 11 ‐, (R)‐11 ‐, and 12 , which present the imidazolidin‐4‐one core and belong to a class of compounds interesting for their biological activities. All the final compounds and intermediates were fully characterized. Enantiomeric excesses of homochiral 6 and 11 were determined by capillary electrophoresis analysis using 2‐hydroxypropyl‐β‐cyclodextrin or highly sulfated γ‐cyclodextrin as chiral selectors. J. Heterocyclic Chem., (2010)