Synthesis, characterization, SAR, antioxidant, anti-acetylcholinesterase and anti-butyrylcholinesterase activities of cephradine Schiff bases.

The objective of this study was to deal with the evaluation of 7-(2-(benzylideneamino)-2-(cyclohexa-1,4-dienyl)acetamido)-3-methyl-8-oxo-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid based schiff bases as a new class of enzyme inhibitors. In this connection, a series of Schiff bases of cephradine with substituted aromatic aldehydes was synthesized and characterized using FTIR, 1HNMR and 13CNMR. The in-vitro biological activities including free radical scavenging potential using DPPH assay, acetyl cholinesterase and butyryl cholinesterase inhibition potential were evaluated. Two compounds of the series 1g and 1h were found to be active against AChE whereas no derivative was active against BChE while the whole series showed excellent 1, 1-diphenyl-2-picrylhydrazyl scavenging activity. All the synthesized compounds were found to be non-toxic and present passive gastrointestinal absorption. Furthermore, the study suggests that the synthesized cephradine derivatives exhibit inhibitory potential against different biologically relevant enzyme targets.

Detection of the iron complexes with hydrolysis products of cephalexin and cefradine upon high-performance liquid chromatography/electrospray ionization mass spectrometry analysis.

RATIONALE: Cephalosporins (e.g. cephalexin, cefradine) are a major group of widely used ß-lactam antibiotics. Hydrolysis of the ß-lactam ring is an important reaction (often undesired) which leads to deactivation of ß-lactams. To the best of our knowledge there is no electrospray ionization mass spectrometry (ESI-MS) data reported concerning the products of hydrolysis of cephalosporins. METHODS: The hydrolysis of cephalexin and cefradine was performed in aqueous NaOH solutions. After the process the solutions were analyzed by high-performance liquid chromatography (HPLC)/ESI-MS. The elemental compositions of the ions discussed were confirmed by the accurate mass measurements on a quadrupole time-of-flight (QTOF) mass spectrometer. RESULTS: Unexpectedly, complexes between the hydrolysis products of cephalexin and cefradine (CFLh and CFRh ) and iron cation were detected upon HPLC/ESI-MS analysis, namely the ions [(CFLh -H)2 +Fe]+ and [(CFRh -H)2 +Fe]+ , although iron was not added to the analyzed solutions or to the mobile phase. These ions were found to be very stable in the gas phase. CONCLUSIONS: The detection of the complexes between the hydrolysis products of cephalosporins and iron may have a positive impact on the sensitivity and specificity of HPLC/ESI-MS analyses of the hydrolysis products of some cephalosporins.

Study on the interaction mechanism between cefradine and Chlamydomonas reinhardtii in water solutions under dark condition.

Our research investigated the hormesis effect of cefradine on the specific growth rates (µ) of single-celled algae (Chlamydomonas reinhardtii) from aqueous solutions. We found the specific growth rate of C. reinhardtii slightly increased with cefradine concentrations within the range 0.5-10 mg/L. Effects of algae density, initial solution pH, and temperature on the adsorption batch assays were investigated. The optimum conditions for cefradine adsorption occurred at a density of 5 × 106 algae cells/mL, a solution pH of 7.0, and a temperature of 25.0 °C. A Box-Behnken design was employed to evaluate correlations between influential factors and cefradine adsorption. The results showed a significant interaction between algae density and temperature. The maximum removal rate could reach 50.13% under the optimal conditions. Additionally, the adsorption mechanisms were explored through Langmuir and Freundlich isotherm equations, adsorption kinetics, and thermodynamics. The results suggested that the adsorption process was monolayer, spontaneous, and endothermic with an increase in randomness at the algae-solution interface, which followed a pseudo-second-order model. All the data indicated that the alga performed a better removal capacity in the antibiotic-containing wastewater treatment process. This study lays the groundwork for a better understanding of the interaction mechanism between cefradine and Chlamydomonas reinhardtii in water solutions under dark condition.

Prediction of hydrolysis pathways and kinetics for antibiotics under environmental pH conditions: a quantum chemical study on cephradine.

Understanding hydrolysis pathways and kinetics of many antibiotics that have multiple hydrolyzable functional groups is important for their fate assessment. However, experimental determination of hydrolysis encounters difficulties due to time and cost restraint. We employed the density functional theory and transition state theory to predict the hydrolysis pathways and kinetics of cephradine, a model of cephalosporin with two hydrolyzable groups, two ionization states, two isomers and two nucleophilic attack directions. Results showed that the hydrolysis of cephradine at pH = 8.0 proceeds via opening of the ß-lactam ring followed by intramolecular amidation. The predicted rate constants at different pH conditions are of the same order of magnitude as the experimental values, and the predicted products are confirmed by experiment. This study identified a catalytic role of the carboxyl group in the hydrolysis, and implies that the carboxyl group also plays a catalytic role in the hydrolysis of other cephalosporin and penicillin antibiotics. This is a first attempt to quantum chemically predict hydrolysis of an antibiotic with complex pathways, and indicates that to predict hydrolysis products under the environmental pH conditions, the variation of the rate constants for different pathways with pH should be evaluated.

Adsorption mechanism of cefradine on three microplastics: A combined molecular dynamics simulation and density functional theory calculation study.

Microplastics and antibiotics are receiving increasing attention as two emerging pollutants in the aquatic ecosystem. The absorption of antibiotics by microplastics can potentially intensify their impact on marine organisms and human health. However, the detailed mechanisms underlying this interaction remain to be elucidated. Through molecular dynamics (MD) simulations and density functional theory (DFT) calculations, this study investigated the adsorption of cefradine (CED) onto three typical microplastics (MPs)-polyethylene (PE), polypropylene (PP), and polyamide (PA). The results of the molecular dynamics simulations showed that the interaction energy between CED and microplastics followed the order of PA-CED > PP-CED > PE-CED, indicating that PA microplastics had the highest adsorption capacity for CED antibiotics. The total energy contribution of the microplastics-cefradine (MPs-CED) systems suggested that the van der Waals and electrostatic interactions were the two primary mechanisms for the adsorption of CED by these three microplastics. In DFT calculations, the adsorption of CED on PA was found to be significantly influenced by both electrostatic and van der Waals effects, while the main driving force in the adsorption of PE and PP is van der Waals effect. In addition, IGMH analysis and AIM topological analysis confirmed that the adsorption of CED on PA relied heavily on the synergistic effect of hydrogen bonding and the van der Waals effect. The findings of this study validate the results obtained from molecular dynamics simulations, laying a foundation for a comprehensive exploration of the interaction mechanisms between microplastics and organic pollutants by integrating MD simulations and DFT calculations.

Structures of cefradine dihydrate and cefaclor dihydrate from DFT-D calculations.

The crystal structure of cefradine dihydrate, C16H19N3O4S·2H2O, is considered in the pharmaceutical sciences to be the epitome of an isolated-site hydrate. The structure from single-crystal X-ray data was described in 1976, but atomic coordinates were not published. The atomic coordinates are determined here by combining the information available from the published single-crystal data with a dispersion-corrected density functional theory (DFT-D) method that has been validated to reproduce molecular crystal structures very accurately. Additional proof for the correctness of the structure comes from comparison with cefaclor dihydrate, C15H14ClN3O4S·2H2O, which is isomorphous and for which more complete single-crystal data are available. H-atom positions have not previously been published for either compound. The DFT-D calculations confirm that both cefradine and cefaclor are present in the zwitterionic form in the two dihydrate structures. A potential ambiguity concerning the orientation of the cyclohexadienyl ring in cefradine dihydrate is also clarified, and on the basis of the calculated energies it is shown that disorder should not be expected at room temperature. The DFT-D methods can be applied to recover full structural data in cases where only partial information is available, and where it may not be possible or desirable to obtain new experimental data.

[Feasibility of the extended application of near infrared universal quantitative models].

Construction of a successful near infrared analysis model is a complex task. It spends a lot of manpower and material resources, and is restricted by sample collection and model optimization. So it is important to study on the extended application of the existing near infrared (NIR) models. In this paper, cephradine capsules universal quantitative model was used as an example to study on the feasibility of its extended application. Slope/bias correction and piecewise direct standardization correction methods were used to make the universal model to fit to predict the intermediates in manufacturing processes of cephradine capsules, such as the content of powder blend or granules. The results showed that the corrected NIR universal quantitative model can be used for process control although the results of the model correction by slope/bias or piecewise direct standardization were not as good as that of model updating. And it also indicated that the model corrected by slope/bias is better than that by piecewise direct standardization. Model correction provided a new application for NIR universal models in process control.

Study on the binding behavior of lysozyme with cephalosporin analogues by fluorescence spectroscopy.

It was first found that the intrinsic fluorescence of lysozyme at 340 nm can be quenched by cephalosporin analogues through the static quenching and non-radiative energy transferring procedure. In the acetate buffer solution with pH 7.0 and 298 K, the quenching fluorescence intensity was in a good linearity over the concentration of drugs in the range of 1-100 micromol L(-1), 0.1-100 micromol L(-1), 0.5-100 micromol L(-1) and 0.05-100 micromol L(-1) for cefradine, cefuroxime, cefotaxime and ceftriaxone, respectively. The quenching ability or the binding ability of the studied drugs followed the pattern: ceftriaxone > cefotaxime > cefuroxime > cefradine, which was close to the order of their antibacterial ability. The binding parameters including the association constant and the number of binding potential point were calculated at different temperatures (288, 298 and 308 K), and thermodynamic parameters DeltaH degrees, DeltaS degrees and DeltaG degrees were given. The binding mode of lysozyme with cephalosporins showed that the hydrophobic effect might play a major role. The binding distance between cephalosporin and tryptophan residue in lysozyme was obtained. The results provided the quantitative information for the binding of cephalosporin to lysozyme, and it was suggested that the drugs probably bound to the active site near Trp62 in lysozyme.

Controlled release of cephradine by biopolymers based target specific crosslinked hydrogels.

The novel silane crosslinked (TEOS) hydrogels based on eco-friendly biodegradable chitosan/guargum were prepared by blending with PEG to develop pH sensitive hydrogels (CGP) and achieved its hydrophilicity and target specificity for controlled release of drug. The crosslinker amount was varied to analyze its effect on the hydrogel properties and were characterized using FTIR, SEM, TGA, swelling studies (water, buffer and ionic solution) and in-vitro release of cephradine (CED). FTIR confirmed the presence of characteristic peaks and crosslinking between the components while SEM images showed the formation of clear micro- and macro-pores. The swelling behavior in water showed that compared to the controlled hydrogel, the crosslinked hydrogels revealed more swelling but a decrease in swelling with further increase in the amount of crosslinker was observed. The hydrogels showed low swelling at basic and neutral pH while maximum swelling was observed at acidic pH. This pH response made these hydrogels an ideal candidate for injectable controlled release. The CED was loaded on hydrogels and its release mechanism was studied in PBS, SGF and SIF which revealed that out of all hydrogels (CGP100, CGP150, CGP200 and CGP250), CGP100 has shown CED release of 85% in 130 min in PBS and 82.4% in SIF.

Cephradine antacids interaction studies.

The present work comprises of interaction studies of cephradine with antacids. Cephradine is included among the first generation cephalosporin, which is active against a wide range of Gram positive and Gram-negative bacteria including penicillinase-producing staphylococci. Since the presence of complexing ligand may affect the bioavailability of a drug in blood or tissues, therefore, in order to study the probable interaction of cephradine with antacids all the reaction conditions were simulated to natural environments. Antacids are commonly used in patients complaining of GI irritations. The behavior of cephradine in presence of seven antacids i.e., simethicone, magaldrate, magnesium carbonate, magnesium hydroxide, magnesium trisilicate, sodium bicarbonate and aluminium hydroxide was studied by using standard dissolution apparatus. Cephradine was monitored both by UV and by high performance liquid chromatography. The results revealed that antacids containing polyvalent cations retarded the in vitro availability of cephradine. Moreover, these studies indicated that cephradine was strongly adsorbed on antacids; magnesium trisilicate and simeco tablets (powdered) exhibited relatively higher adsorption capacities.

Computational design of cephradine synthase in a new scaffold identified from structural databases.

Computational enzyme design exhibits excellent performance for identifying potential scaffolds from structural databases and creating new enzymatic catalysts from naught. Using the active site-matching algorithm ProdaMatch, we identified a new scaffold cocaine esterase from Rhodococcus sp. that showed modest activity (kcat/Km = 0.018 M-1 s-1) towards the hydrolysis of ß-lactam antibiotic cephradine. The identified cocaine esterase scaffold afforded low sequence identity (<30%) with the known ß-lactam synthases, such as penicillin G acylase or α-amino acid ester hydrolase, and was able to catalyze the condensation reaction between d-dihydrophenylglycine methyl ester and 7-aminodesacetoxycephalosporanic acid to produce cephradine via a kinetically controlled synthesis. By virtue of the computational enzyme design protocol, hundreds of sequences were predicted in the cocaine esterase scaffold to promote the catalytic activity towards the hydrolytic reaction of cephradine. Moreover, a single mutant (F261T) was experimentally confirmed to have improved the catalytic efficiency by ten times (kcat/Km = 0.193 M-1 s-1), indicating that the novel scaffold cocaine esterase may be potentially redesigned to become an industrially useful cephradine synthase.

Controlled drug delivery system based on magnetic hollow spheres/polyelectrolyte multilayer core-shell structure.

A novel controlled drug delivery system was fabricated by coating chitosan/PAA multilayer onto magnetic hollow spheres via a "Layer-by-Layer" (LBL) assembly approach. Cefradine was used as a model drug to evaluate the drug release characteristics of this core-shell hollow structure and the results show that it exhibits a sustained release of the drug and the release rate can be regulated by the pH environment of release medium. It is believed that this core-shell hollow structure, which combines the advantage of controlled delivery as well as magnetic targeting, has commendable potential in drug delivery therapeutics.

Crystal form of cephradine.

Four crystal forms of cephradine were isolated by recrystallization and characterized by powder X-ray diffractometry, differential scanning calorimetry, and thermogravimetric analysis. The dissolution patterns of four crystal forms of cephradine were studied in water at 37 +/- 0.5 degrees C, 90 rpm for 120 min. The amount dissolved at 120 min was highest for Form I (100%), followed by Form 3 (98.9%), Form 4 (77.83%), and Form 2 (75.55%). After storage for two months at 0% RH (silica gel, 20 degrees C), 52% RH (saturated solution of Na2Cr2O7 x 2H2O/ 20 degrees C), and 95% RH (saturated solution of Na2HPO4/20 degrees C), none of the crystal forms showed transformation.

Preparation and characterization of uniform nanosized cephradine by combination of reactive precipitation and liquid anti-solvent precipitation under high gravity environment.

In this work, a novel direct method, which was combined with reactive precipitation and liquid anti-solvent precipitation under high gravity environment, had been developed to prepare nanosized cephradine with narrow particle size distribution. Compared with commercial crude cephradine, the prepared cephradine showed a significant decrease in particle size, a significant increase in the specific surface area and shorter dissolving time when used for injection. The characteristic particle size was between 200-400 nm. The specific surface area increased from 2.95 to 10.87 m2/g after micronization. When the amount of L-arginin decreased from 0.25 to 0.18 g, the mixture of nanosized cephradine and L-arginine could still dissolve in 1 min. The X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) analysis indicated that the physical characteristics and molecular states remained unchanged after the recrystallization process. This method had potential application in industrial fields because of its low cost, efficient processing and the ease of scaling-up.

A model assessment of the importance of direct photolysis in the photo-fate of cephalosporins in surface waters: Possible formation of toxic intermediates.

The direct and indirect photodegradation of six cephalosporins was predicted using a photochemical model, on the basis of literature values of photochemical reactivity. Environmental photodegradation would be important in surface water bodies with depth ⩽ 2-3m, and/or in deeper waters with low values of the dissolved organic carbon (DOC ⩽ 1 mg C L(-1)). The half-life times would range from a few days to a couple of weeks in summertime. In deeper and higher-DOC waters and/or in different seasons, hydrolysis could prevail over photodegradation. The direct photolysis of cephalosporins is environmentally concerning because it is known to produce toxic intermediates. It would be a major pathway for cefazolin, an important one for amoxicillin and cefotaxime and, at pH<6.5, for cefapirin as well. In contrast, direct photolysis would be negligible for cefradine and cefalexin. The DOC values would influence the fraction of photodegradation accounted for by direct photolysis in shallow water, to a different extent depending on the role of sensitisation by the triplet states of chromophoric dissolved organic matter.

Study of the nucleation and growth of antibiotic labeled Au NPs and blue luminescent Au8 quantum clusters for Hg(2+) ion sensing, cellular imaging and antibacterial applications.

Herein, we report a detailed experimental study supported by DFT calculations to understand the mechanism behind the synthesis of cefradine (CFD--an antibiotic) labeled gold nanoparticles (Au NPs) by employing CFD as both a mild reducing and capping agent. The analysis of the effect of growth conditions reveals that a higher concentration of HAuCl4 results in the formation of an increasing fraction of anisotropic structures, higher temperature leads to the formation of quasi-spherical particles instead of anisotropic ones, and larger pH leads to the formation of much smaller particles. The cyclic voltammetry (CV) results show that when the pH of the reaction medium increases from 4 to 6, the reduction potential of CFD increases which leads to the synthesis of nanoparticles (in a pH 4 reaction) to quantum clusters (in a pH 6 reaction). The MALDI-TOF mass spectrometry results of supernatant of the pH 6 reaction indicate the formation of [Au8(CFD)2S6] QCs which show fluorescence at ca. 432 nm with a Stokes shift of ca. 95 nm. The blue luminescence from Au8 QCs was applied for sensing of Hg(2+) ions on the basis of an aggregation-induced fluorescence quenching mechanism and offers good selectivity and a high sensitivity with a limit of detection ca. 2 nM which is lower than the detection requirement of 10 nM by the U.S. EPA and 30 nM by WHO for drinking water. We have also applied the sensing probe to detect Hg(2+) ions in bacterial samples. Further, we have investigated the antibacterial property of as-synthesized Au NPs using MIC, growth curve and cell survival assay. The results show that Au NPs could reduce the cell survival very efficiently rather than the cell growth in comparison to the antibiotic itself. The scanning electron microscopy study shows the degradation and blebbing of the bacterial cell wall upon exposure with Au NPs which was further supported by fluorescence microscopy results. These Au NPs did not show reactive oxygen species generation. We believe that the bacterial cytotoxicity is due to the direct contact of the Au NPs with bacterial cells.

Preparation and characterization of porous hollow silica nanoparticles for drug delivery application.

Porous hollow silica nanoparticles (PHSNP) with a diameter of 60-70 nm and wall thickness of approximately 10nm were synthesized by using CaCO(3) nano-particles as the inorganic template. The characterization of PHSNP by TEM and BET indicated that PHSNP were uniform spherical particles with good dispersion, and had a specific surface area of 867 m(2)/g. The as-synthesized PHSNP were subsequently employed as drug carrier to investigate in vitro release behavior of cefradine in simulated body fluid. UV-spectrometry and TG analyses were performed to determine the amount of cefradine entrapped in the carrier. The BJH pore size distribution of PHSNP before and after entrapping cefradine was examined. Cefradine release profile from PHSNP followed a three-stage pattern and exhibited a delayed release effect.

Capillary electrophoresis study on the micellization and critical micelle concentration of sodium dodecyl sulfate. Influence of solubilized solutes.

The influence of solubilized solutes on the micellization and critical micelle concentration (CMC) of sodium dodecyl sulfate (SDS) were investigated by means of capillary electrophoresis (CE). Three different structural types of test solutes, including chloropyridines. chlorophenols and cephalosporins with different binding strength to SDS micelles, were selected in this study. The variations of the effective electrophoretic mobility of these solutes as a function of SDS concentration in the premicellar and micellar regions were analyzed. Interestingly, the results indicate that, in the presence of these solubilized solutes, the micellization of SDS may occur over a range of SDS concentration, with the aggregate size increasing over this range. Depending on the nature of solubilized solutes and the extent of the interactions between solubilized solutes and SDS micelles, the CMC value of SDS may vary significantly. The incorporation of solubilized solutes into SDS micelles to form mixed micelles is proposed to interpret the migration behavior of solubilized solutes in CE.

Treatment of anti-osmotic drug based pharmaceutical effluent in an upflow anaerobic fluidized bed system.

During the production of Cephradine (a main constituent of anti-osmotic drug) a large quantity of concentrated effluent was produced. The main polluting compounds in this effluent are osmotic drug, acetic acid and ammonia. The feasibility of using a fluidized bed reactor under anaerobic condition with bioaugmentation to treat anti-osmotic drug based pharmaceutical effluent was evaluated. The main objective of the study was to show that bioaugmentation could be used to promote biological treatment to applications where conventional operation might be difficult or unfavourable. The effluent with chemical oxygen demand (COD) of 14000-18000 mg/l was treated in a fluidized bed reactor with a hydraulic retention time of 3-12 h. The reactor was unable to maintain consistent removal in conventional mode of operation due to an inability to retain and grow biomass. The COD reduction (%) after inoculation from a sequencing batch reactor was related to influent concentration, mass of inoculum and hydraulic retention time characterized by calculating the initial food to microorganism ratio. The role of volatile fatty acid (VFA) as cosubstrate was assessed with respect to COD reduction (%). Continuous COD reduction (%) attained a maximum value of 88.5% using bioaugmentation through periodic addition of acclimated cells every 2 days with 30-73.2 g of cells from an off-line enricher reactor.

Novel controlled drug delivery system for multiple drugs based on electrospun nanofibers containing nanomicelles.

This research described a novel composite electrospun nanofibers, which were consisted of MPEG-b-PLA micelles, chitosan, and PEO, realizing controlled release of both hydrophobic and hydrophilic drugs. 5-FU and Cefradine used as model drugs were successfully loaded in the nanofibers. The in vitro studies showed there was a low initial burst release of 5-FU from micelles-loaded nanofibers, and the final release proportion was about 91.4% after continually releasing for 109 h. In vitro cytotoxicity studies revealed that 5-FU-loaded nanofibers restrained HepG-2 cells efficiently, and the cell viability was 45.9% after three days of cultivation in solutions containing micelles-loaded nanofibers with 21.6 µg 5-FU. All results suggested that micelles-loaded nanofibers with two kinds of drugs can be used as an effective controlled drug delivery vehicle and may have a bright future in cancer chemotherapy or clinical treatments.