RESUMO
FT-IR/ATR analytical technique is one of the most applicable techniques worldwide. It is closely associated with easy-to-use equipment, rapid analysis, and reliable results. This study reports the simultaneous qualitative and quantitative analysis of two active pharmaceutical ingredients (APIs), of a piperacillin and tazobactam formulation using a film formation method. This method requires film formation on the ATR crystal, resulting from solvent evaporation of a small amount of liquid sample. Good contact between the film and the crystal led to the identification of both APIs, although tazobactam was of low content in the formulation mixture. The quantification of the APIs in the commercial mixture was also achieved, using a single calibration line with a correlation coefficient equal to 0.999, not only after film formation but also in the initial dry formulation before reconstitution. The present spectroscopic technique combined with the proposed relatively simple sample treatment outweighs chromatographic protocols already applied, which require specialized staff and are costly, time-consuming, and not environmentally friendly. Taking all the above into consideration, it turns out that such an approach has the potential to be used for off-line quality control procedures in manufacture or, in terms of portable equipment and automated software, anywhere for on-site analysis, even in a hospital workflow.
Assuntos
Piperacilina/química , Tazobactam/química , Calibragem , Química Farmacêutica/métodos , Estudos de Avaliação como Assunto , Preparações Farmacêuticas/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodosRESUMO
In this study, drug nanocarriers were designed using linear copolymers with different contents of cholinium-based ionic liquid units, i.e., [2-(methacryloyloxy)ethyl]trimethylammonium chloride (TMAMA/Cl: 25, 50, and 75 mol%). The amphiphilicity of the copolymers was evaluated on the basis of their critical micelle concentration (CMC = 0.055-0.079 mg/mL), and their hydrophilicities were determined by water contact angles (WCA = 17°-46°). The chloride anions in the polymer chain were involved in ionic exchange reactions to introduce pharmaceutical anions, i.e., p-aminosalicylate (PAS-), clavulanate (CLV-), piperacillin (PIP-), and fusidate (FUS-), which are established antibacterial agents for treating lung and respiratory diseases. The exchange reaction efficiency decreased in the following order: CLV- > PAS- > PIP- >> FUS-. The hydrophilicity of the ionic drug conjugates was slightly reduced, as indicated by the increased WCA values. The major fraction of particles with sizes ~20 nm was detected in systems with at least 50% TMAMA carrying PAS or PIP. The influence of the drug character and carrier structure was also observed in the kinetic profiles of the release processes driven by the exchange with phosphate anions (0.5-6.4 µg/mL). The obtained polymer-drug ionic conjugates (especially that with PAS) are promising carriers with potential medical applications.
Assuntos
Antituberculosos/administração & dosagem , Colina/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Líquidos Iônicos/química , Polímeros/química , Ácido Aminossalicílico/química , Ânions/química , Antituberculosos/química , Antituberculosos/farmacocinética , Ácido Clavulânico/química , Liberação Controlada de Fármacos , Ácido Fusídico/química , Interações Hidrofóbicas e Hidrofílicas , Micelas , Piperacilina/química , Espectroscopia de Prótons por Ressonância Magnética/métodos , Espectrofotometria/métodosRESUMO
BACKGROUND: The mortality rate of patients with a drug-resistant bacterial infection is high, as are the associated treatment costs. To overcome these issues, optimization of the available therapeutic options is required. Beta-lactams are time-dependent antibiotics and their efficacy is determined by the amount of time the free concentration remains above the minimum inhibitory concentration. Therefore, the aim of this study was to assess the extent and variability of protein binding for meropenem, cefepime, and piperacillin. METHODS: Plasma samples for the analysis of meropenem, cefepime, and piperacillin were collected from patients admitted to a tertiary care hospital as part of the standard care. The bound and unbound drug fractions in the samples were separated by ultrafiltration. Validated liquid chromatography-tandem mass spectrometry assays were used to quantify the total and free plasma concentrations, and the protein binding was determined. RESULTS: Samples from 95 patients were analyzed. The median (range) age of patients was 56 years (17-87) and the median (range) body mass index was 25.7 kg/m (14.7-74.2). Approximately 59% of the patients were men. The median (range) unbound fraction (fu) was 62.5% (41.6-99.1) for meropenem, 61.4% (51.6-99.2) for cefepime, and 48.3% (39.4-71.3) for piperacillin. In the bivariate analysis, as the total meropenem concentration increased, the fu increased (r = 0.37, P = 0.045). A decrease in piperacillin fu was observed as the albumin concentration increased (r = -0.56, P = 0.005). CONCLUSIONS: The average fu values were lower than those reported in the literature. There was also a large variability in fu; hence, it should be considered when managing patients administered with these drugs through direct measurements of free drug concentrations.
Assuntos
Antibacterianos/metabolismo , Cefepima/metabolismo , Meropeném/metabolismo , Piperacilina/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/sangue , Antibacterianos/química , Infecções Bacterianas/tratamento farmacológico , Cefepima/sangue , Cefepima/química , Monitoramento de Medicamentos , Feminino , Humanos , Masculino , Meropeném/sangue , Meropeném/química , Pessoa de Meia-Idade , Piperacilina/sangue , Piperacilina/química , Ligação Proteica , Adulto JovemRESUMO
Piperacillin is a broad spectrum beta-lactam antibiotic used in combination with tazobactam for hospital-related bacterial infections. The reconstituted solutions must respect the sub-visible and visible particles specifications. It was claimed that the reformulation containing EDTA/sodium citrate was able to control the formation of an insoluble impurity responsible for the formation of particulate matter observed using Ringer Lactate as diluent. The nature of the impurities formed during the degradative process of piperacillin/tazobactam combination has been herein investigated, by exploring the effect of added excipients and pH variations. The exact structure of the isolated dimeric impurity, the penicilloic acid-piperacillin dimer, was determined through complete characterization, allowing to propose a novel degradative general pathway for beta-lactam antibiotics. The presence of EDTA resulted unnecessary to contain the formation of the insoluble impurity, since the use of sodium citrate alone allowed to avoid this drawback. Finally, the proposed mechanism was successfully applied to the design of a novel, easy and high purity procedure for the synthesis of the acetylated penicilloic acid, known related substance of piperacillin.
Assuntos
Antibacterianos/química , Penicilinas/química , Combinação Piperacilina e Tazobactam/química , Piperacilina/química , Tazobactam/química , Infecções Bacterianas/tratamento farmacológico , Quimioterapia Combinada/métodos , Humanos , Testes de Sensibilidade Microbiana/métodos , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/químicaRESUMO
In intensive care, beta-lactams can be reconstituted in 50 mL polypropylene syringes with NaCl 0.9 % and administered for 8 to 12 h at various concentrations with motor-operated syringe pumps. The feasibility and/or the stability of these antibiotic therapies are often poorly known by clinicians. The purpose of this study was to determine the stability of seven antipyocyanic beta-lactam antibiotics and cilastatin under real-life conditions. Stability indicating HPLC methods allowing quantification in pharmaceutical preparations and subsequent stability studies were performed. The stability studies showed that continuous infusion of piperacillin/tazobactam 80/10 mg/mL, of cefepime 20 and 40 mg/mL and of aztreonam 40 and 120 mg/mL can be used over 12 h. Moreover, continuous infusion of cefepime 120 mg/mL can be used over 10 h, whereas meropenem 10 and 20 mg/mL and ceftazidime 40 mg/mL remained stable only over 8 h, and meropenem 40 mg/mL was significantly degraded after 6 h. Finally, imipenem/cilastatin 5/5 mg/mL and piperacillin/tazobactam 320/40 mg/mL should not be used as continuous infusion. These data allow the establishment of protocols of administration of antipyocyanic beta-lactams by continuous infusion. Some of them are not appropriate to this mode of administration (imipenem/cilastatin, piperacillin/ tazobactam 320/40 mg/mL) or must be avoided if possible (ceftazidime 40 mg/mL).
Assuntos
Antibacterianos/química , beta-Lactamas/antagonistas & inibidores , Aztreonam/química , Cefepima/química , Ceftazidima/química , Cilastatina/química , Combinação Imipenem e Cilastatina/química , Imipenem/química , Meropeném/química , Piperacilina/química , Combinação Piperacilina e Tazobactam/química , Tazobactam/químicaRESUMO
ßLactam antibiotics are among the most widely used antibiotics in human medicine and their effects on the aquatic environment - concerning bacterial resistance - are controversially discussed. This study focused on the photolysis of the four ßlactam antibiotics - amoxicillin, ampicillin, penicillin V and piperacillin - under simulated environmental conditions. It was observed that all investigated ßlactam antibiotics are photolytically degradable by simulated sunlight (1â¯kW/m2) with half-lives between 3.2 and 7.0â¯h. Structure elucidation of transformation products performed with liquid chromatography coupled to high resolution mass spectrometry showed that the hydrolysis of the ßlactam ring is the primary transformation reaction, followed by the elimination of carboxylic and dimethyl thiazolidine carboxylic acid. Growth inhibition tests on Bacillus subtilis showed the loss of bactericide activity of irradiated solutions of amoxicillin, ampicillin and piperacillin, suggesting the transformation of the ßlactam ring is responsible for the antibiotic effect. In contrast, the solutions of penicillin V did not show any decline of the antibacterial activity after photolytic degradation, probably due to the formation of still active epimers.
Assuntos
Antibacterianos/química , Fotólise , Eliminação de Resíduos Líquidos , Poluentes Químicos da Água/química , beta-Lactamas/química , Amoxicilina/química , Amoxicilina/farmacologia , Ampicilina/química , Ampicilina/farmacologia , Antibacterianos/farmacologia , Biodegradação Ambiental , Penicilina V/química , Penicilina V/farmacologia , Piperacilina/química , Piperacilina/farmacologia , Luz Solar , beta-Lactamas/farmacologiaRESUMO
Carbapenems are "last resort" ß-lactam antibiotics used to treat serious and life-threatening health care-associated infections caused by multidrug-resistant Gram-negative bacteria. Unfortunately, the worldwide spread of genes coding for carbapenemases among these bacteria is threatening these life-saving drugs. Metallo-ß-lactamases (MßLs) are the largest family of carbapenemases. These are Zn(II)-dependent hydrolases that are active against almost all ß-lactam antibiotics. Their catalytic mechanism and the features driving substrate specificity have been matter of intense debate. The active sites of MßLs are flanked by two loops, one of which, loop L3, was shown to adopt different conformations upon substrate or inhibitor binding, and thus are expected to play a role in substrate recognition. However, the sequence heterogeneity observed in this loop in different MßLs has limited the generalizations about its role. Here, we report the engineering of different loops within the scaffold of the clinically relevant carbapenemase NDM-1. We found that the loop sequence dictates its conformation in the unbound form of the enzyme, eliciting different degrees of active-site exposure. However, these structural changes have a minor impact on the substrate profile. Instead, we report that the loop conformation determines the protonation rate of key reaction intermediates accumulated during the hydrolysis of different ß-lactams in all MßLs. This study demonstrates the existence of a direct link between the conformation of this loop and the mechanistic features of the enzyme, bringing to light an unexplored function of active-site loops on MßLs.
Assuntos
Antibacterianos/química , Ceftazidima/química , Imipenem/química , Meropeném/química , Zinco/química , beta-Lactamases/química , Sequência de Aminoácidos , Antibacterianos/metabolismo , Domínio Catalítico , Cefepima/química , Cefepima/metabolismo , Cefotaxima/química , Cefotaxima/metabolismo , Ceftazidima/metabolismo , Clonagem Molecular , Cristalografia por Raios X , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Imipenem/metabolismo , Cinética , Meropeném/metabolismo , Modelos Moleculares , Piperacilina/química , Piperacilina/metabolismo , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Engenharia de Proteínas , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Zinco/metabolismo , Resistência beta-Lactâmica , beta-Lactamases/genética , beta-Lactamases/metabolismoRESUMO
INTRODUCTION: Liquid chromatography coupled to atmospheric pressure ionization tandem mass spectrometry (LC-ESI-MS/MS) is currently considered the reference method for quantitative determination of urinary free cortisol (UFC). One of the major drawbacks of this measurement is a particular form of matrix effect, conventionally known as ion suppression. MATERIALS AND METHODS: We describe here the case of a 66-year-old-patient referred to the daily service of general medicine for intravenous antibiotic administration due to a generalized Staphylococcus aureus infection and for routine 24 hours UFC monitoring in the setting of glucocorticoid replacement therapy. RESULTS: The observation of 10-fold decrease of internal standard of cortisol signal led us to hypothesize the presence of an ion suppression effect due to a co-eluting endogenous compound. Screening analysis of tandem mass spectrometry (MS/MS) spectra of the interfering molecule, along with in vitro confirmation analyses, were suggestive of the presence of high concentration of piperacillin. The problem was then easily solved with minor modifications of the chromatographic technique. CONCLUSIONS: According to our findings, antibiotic therapy with piperacillin/tazobactam should be regarded as an important interference in UFC assessment, which may potentially affect detection capability, precision and accuracy of this measurement. This case report emphasizes that accurate anamnesis and standardization of all phases of urine collection are essential aspects for preventing potential interference in laboratory testing.
Assuntos
Antibacterianos/uso terapêutico , Hidrocortisona/urina , Piperacilina/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Espectrometria de Massas em Tandem , Idoso , Antibacterianos/química , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Íons/química , Piperacilina/químicaRESUMO
Background: Elastomeric pumps can be used for the continuous administration of antimicrobials in the outpatient setting. A potentially limiting factor in their use is the stability of antimicrobials. Objectives: To investigate under real-life conditions the temperature variations of antibiotic solutions contained in elastomeric pumps, and to examine under such conditions the stability of five antibiotics. Methods: Healthy volunteers carried the elastomeric pumps in carry pouches during their daily activities. A thermologger measured the temperatures every 15â min over 24â h. Antibiotic concentrations were measured by HPLC coupled to tandem MS. Results: During daytime, the temperature of solutions in the pumps increased steadily, warming to >30°C. During the night, when the pumps were kept attached to the waist, the temperatures reached up to 33°C. The use of white carry pouches avoided excessive temperature increases. Over seven experiments, cefazolin, cefepime, piperacillin and tazobactam were found to be stable over 24â h. Flucloxacillin showed a mean decrease in concentration of 11% ( P = 0.001). Conclusions: Real-life situations can cause significant temperature rises in elastomeric pumps, thereby potentially increasing the risk of antibiotic degradation. Patients should be instructed to avoid situations causing excessive temperature increases. Despite these temperature variations, cefazolin, cefepime, piperacillin and tazobactam were found to be stable over 24â h. A moderate degradation was noticed for flucloxacillin, albeit most probably not to an extent that might impair anti-infective efficacy.
Assuntos
Antibacterianos/química , Estabilidade de Medicamentos , Cefazolina/química , Cefazolina/metabolismo , Cefepima , Cefalosporinas/química , Elastômeros , Feminino , Floxacilina/química , Voluntários Saudáveis , Humanos , Bombas de Infusão , Masculino , Piperacilina/química , Polímeros , TemperaturaRESUMO
Objective. The purpose of this study is to collect data on epidemiology, microbiology, and outcome of VAP in our ICUs for reevaluation of the therapeutic strategies. Methods. This retrospective study involved all adult patients, 15 years of age or older, diagnosed with VAP in multidisciplinary ICUs at Hamad General Hospital between January 2010 and December 2012. Results. A total of 106 patients were enrolled. The mean incidence of VAP was 5.0 per 1000 ventilator-days. It was predominant among younger age group (<60 years), male patients (80.2%), and trauma ICU admissions (49.0%). The most common comorbidity was hypertension (34%) and polytrauma (36.8%) was the most frequent admission diagnosis. 30-day mortality was 23.6% and it was significantly higher in ≥60 years age group, female gender, patients with diabetes mellitus, hypertension, chronic respiratory disease, ≥1 comorbidity, and poor functional status, smokers, medical and surgical ICU admissions, and patients with previous stay in medical/surgical wards, inappropriate empirical therapy, and admission diagnosis of respiratory failure. Gram-negative bacilli were the most frequent respiratory specimen isolates, Pseudomonas spp. being the most common. Majority of our Acinetobacter isolates were multidrug resistant. Conclusion. The incidence of VAP in our ICUs was low. Higher mortality rates were observed in certain subgroup of patients. Resistance to commonly used antimicrobials is likely to require reevaluation of the therapeutic strategies at our institution.
Assuntos
Cuidados Críticos/métodos , Unidades de Terapia Intensiva , Pneumonia Associada à Ventilação Mecânica/epidemiologia , Pneumonia Associada à Ventilação Mecânica/terapia , Acinetobacter , Adolescente , Adulto , Idoso , Anti-Infecciosos/química , Comorbidade , Coleta de Dados , Farmacorresistência Bacteriana Múltipla , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/química , Piperacilina/química , Pseudomonas , Catar/epidemiologia , Estudos Retrospectivos , Fatores de Risco , Tazobactam , Resultado do Tratamento , Adulto JovemRESUMO
PURPOSE: The compatibility of vancomycin and piperacillin-tazobactam in concentrations typically used in extended-infusion dosing schemes was evaluated. METHODS: Piperacillin-tazobactam was reconstituted and diluted to concentrations of 33.75, 45, 50, 60, 67.5, 80, and 90 mg/mL. Vancomycin was diluted to concentrations of 4-8, 10, and 12 mg/mL. The resultant admixtures were visually observed after preparation against black and white backgrounds each hour between hours 1 through 4 and after 24 hours. Frozen products of each medication and brand-name Zosyn powder for reconstitution also were studied. Each combination of products and concentrations was tested for precipitation using simulated Y-site administration. Absorbance and microscopic analyses were performed to discern less perceptible incompatibilities in combinations that did not result in visual precipitation. Changes in absorbance were evaluated using two-way repeated-measures analysis of variance with post hoc Bonferroni corrections. RESULTS: No tested concentrations of piperacillin-tazobactam showed precipitations with vancomycin up to concentrations of 7 mg/mL. Piperacillin-tazobactam 80-90 mg/mL formed reversible precipitation with vancomycin 8 mg/mL. All tested concentrations of piperacillin-tazobactam formed a reversible precipitate with vancomycin 10 mg/mL. Irreversible precipitation was noted with all combinations of piperacillin-tazobactam and vancomycin 12 mg/mL. No significant changes in absorbance analyses were identified for all tested piperacillin-tazobactam concentrations and vancomycin 4-10 mg/mL compared with 0.9% sodium chloride injection (p > 0.05). Similar results were observed using frozen preparations and brand-name Zosyn. CONCLUSION: Visual, microscopic, and absorbance analyses showed no evidence of incompatibility when piperacillin-tazobactam 33.75-90 mg/mL was combined with vancomycin ≤7 mg/mL. Reversible and irreversible precipitates formed when piperacillin-tazobactam was combined with vancomycin ≥8 mg/mL.
Assuntos
Antibacterianos/química , Ácido Penicilânico/análogos & derivados , Vancomicina/química , Antibacterianos/administração & dosagem , Precipitação Química , Composição de Medicamentos/métodos , Incompatibilidade de Medicamentos , Humanos , Ácido Penicilânico/administração & dosagem , Ácido Penicilânico/química , Piperacilina/administração & dosagem , Piperacilina/química , Combinação Piperacilina e Tazobactam , Cloreto de Sódio/química , Fatores de Tempo , Vancomicina/administração & dosagemRESUMO
A green, novel, rapid, accurate and reliable capillary zone electrophoresis method was developed and validated for the simultaneous determination of piperacillin, tazobactam and cefepime in pharmaceutical preparations. Separation was carried out using fused silica capillary (50 µm i.d. × 48.6 cm and 40.2 cm detection length) and applied potential of 20 kV (positive polarity) and a running buffer containing 15 m m sodium borate buffer adjusted to pH 9.3 with UV detection at 215 nm. Amoxicillin was used as an internal standard. The method was suitably validated according to International Conference on Harmonization guidelines. The method showed good linearity in the ranges of 10-100, 20-400 and 10-400 µg/mL with limits of quantitation of 1.87, 3.17 and 6.97 µg/mL and limits of detection of 0.56, 0.95 and 2.09 µg/mL for tazobactam, piperacillin and cefepime, respectively. The proposed method was successfully applied for the analysis of these drugs in their synthetic mixtures and co-formulated injection vials. The method was extended to the in vitro determination of the two drugs in spiked human plasma. It is considered a 'green' method as it consumes no organic solvents.
Assuntos
Cefalosporinas/análise , Eletroforese Capilar/métodos , Ácido Penicilânico/análogos & derivados , Piperacilina/análise , Cefepima , Cefalosporinas/sangue , Cefalosporinas/química , Formas de Dosagem , Química Verde , Humanos , Modelos Lineares , Ácido Penicilânico/análise , Ácido Penicilânico/sangue , Ácido Penicilânico/química , Piperacilina/sangue , Piperacilina/química , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , TazobactamRESUMO
Pharmaceutical differences between the reference listed drug (RLD) and generic formulations of piperacillin-tazobactam may impact the reconstitution process for intravenous administration. This study evaluated the RLD against three generic formulations and measured their reconstitution times using a standardized process. The mean (standard deviation [SD]) reconstitution time for one generic formulation was 5.57 (1.49) min, which was 35% to 42% longer (P < 0.002) than that for the RLD and two other formulations. Observable microscopic differences in powder particle morphology may explain these findings.
Assuntos
Antibacterianos/química , Medicamentos Genéricos/química , Ácido Penicilânico/análogos & derivados , Piperacilina/química , Administração Intravenosa/métodos , Química Farmacêutica/métodos , Ácido Penicilânico/química , Combinação Piperacilina e Tazobactam , TazobactamRESUMO
BACKGROUND: Recent studies have demonstrated highly variable blood concentrations of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in critically ill patients with a high incidence of sub-therapeutic levels. Consequently, therapeutic drug monitoring (TDM) of these antibiotics has to be considered, requiring robust and reliable routine analytical methods. The aim of the present work was to develop and validate a multi-analyte ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of the above mentioned antibiotics. METHODS: Sample preparation included a manual protein precipitation step followed by two-dimensional ultra high performance liquid chromatography (2D-UHPLC). Corresponding stable isotope-labeled substances were used as internal standards for all of the analytes, with the exception of tazobactam. The injected sample volume was 7 µL. The run time was 5.0 min. RESULTS: Inaccuracy was ≤8% and imprecision coefficient of variation (CV) was <9% for all analytes. Only minor matrix effects and negligible carry-over was observed. The method was found to be robust during the validation period. CONCLUSIONS: We were able to develop a reliable 2D-UHPLC-MS/MS method addressing analytes with highly heterogeneous physico-chemical properties. The novel assay may be an efficient tool for an optimized process workflow in clinical laboratories for important antibiotics in regards to TDM.
Assuntos
Métodos Analíticos de Preparação de Amostras/métodos , Antibacterianos/sangue , Antibacterianos/isolamento & purificação , Análise Química do Sangue/métodos , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Antibacterianos/química , Automação , Cefepima , Cefalosporinas/sangue , Cefalosporinas/química , Cefalosporinas/isolamento & purificação , Ciprofloxacina/sangue , Ciprofloxacina/química , Ciprofloxacina/isolamento & purificação , Humanos , Isótopos/química , Linezolida/sangue , Linezolida/química , Linezolida/isolamento & purificação , Meropeném , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/sangue , Ácido Penicilânico/química , Ácido Penicilânico/isolamento & purificação , Piperacilina/sangue , Piperacilina/química , Piperacilina/isolamento & purificação , Tazobactam , Tienamicinas/sangue , Tienamicinas/química , Tienamicinas/isolamento & purificação , Fatores de TempoRESUMO
Adverse side effects of drugs are often caused by the interaction of drug molecules to targets other than the intended ones. In this study, we investigated the off-target interactions of some commercially available drugs with human α-thrombin. The drugs used in the study were selected from Super Drug Database based on the structural similarity to a known thrombin inhibitor argatroban. Interactions of these drugs with thrombin were initially checked by in silico docking studies and then confirmed by thrombin inhibition assay using a fluorescence microplate-based method. Results show that the three commonly used drugs piperacillin (anti-bacterial), azlocillin (anti-bacterial), and metolazone (anti-hypertensive and diuretic) have thrombin inhibitory activity almost similar to that of argatroban. The Ki values of piperacillin, azlocillin, and metolazone with thrombin are .55, .95, and .62 nM, respectively. The IC50 values of piperacillin, azlocillin, and metolazone with thrombin are 1.7, 2.9, and 1.92 nM, respectively. This thrombin inhibitory activity might be a reason for the observed side effects of these drugs related to blood coagulation and other thrombin activities. Furthermore, these compounds (drugs) may be used as anti-coagulants as such or with structural modifications.
Assuntos
Antitrombinas/química , Simulação de Acoplamento Molecular , Ácidos Pipecólicos/química , Trombina/química , Antitrombinas/metabolismo , Arginina/análogos & derivados , Azlocilina/química , Azlocilina/metabolismo , Humanos , Cinética , Metolazona/química , Metolazona/metabolismo , Estrutura Molecular , Ácidos Pipecólicos/metabolismo , Piperacilina/química , Piperacilina/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Sulfonamidas , Trombina/metabolismoRESUMO
Piperacillin-tazobactam (PTZ) is known to cause false-positive results in the Platelia Aspergillus enzyme-linked immunoassay (EIA), due to contamination with galactomannan (GM). We tested 32 lots of PTZ and 27 serum specimens from patients receiving PTZ. GM was not detected in the lots of PTZ; one serum specimen (3.7%) was positive. PTZ formulations commonly used in the United States today appear to be a rare cause for false-positive GM results.
Assuntos
Antibacterianos/uso terapêutico , Aspergilose/diagnóstico , Aspergillus/isolamento & purificação , Testes Diagnósticos de Rotina/métodos , Reações Falso-Positivas , Mananas/sangue , Soro/química , Idoso de 80 Anos ou mais , Antibacterianos/química , Contaminação de Medicamentos , Feminino , Galactose/análogos & derivados , Humanos , Técnicas Imunoenzimáticas/métodos , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/química , Ácido Penicilânico/uso terapêutico , Piperacilina/química , Piperacilina/uso terapêutico , Combinação Piperacilina e Tazobactam , Estados UnidosRESUMO
The aim of this work was to determine the concentrations and properties of free radicals in piperacillin, ampicillin, and crystalline penicillin after gamma irradiation. The radicals were studied by electron paramagnetic resonance (EPR) spectroscopy using an X-band spectrometer (9.3 GHz). Gamma irradiation was performed at a dose of 25 kGy. One- and two-exponential functions were fitted to the experimental data, in order to assess the influence of the antibiotics' storage time on the measured EPR lines. After gamma irradiation, complex EPR lines were recorded confirming the presence of a large number of free radicals formed during the irradiation. For all tested antibiotics, concentrations of free radicals and parameters of EPR spectra changed with storage time. The results obtained demonstrate that concentration of free radicals and other spectroscopic parameters can be used to select the optimal parameters of radiation sterilization of ß-lactam antibiotics. The most important parameters are the constants τ (τ (1(A),(I)) and τ (2(A),(I))) and K (K (0(A),(I)), K (1(A),(I)), K (2(A),(I))) of the exponential functions that describe free radicals decay during samples storage.
Assuntos
Antibacterianos/química , Radicais Livres/análise , Raios gama , Ampicilina/química , Microscopia Eletrônica de Transmissão por Filtração de Energia , Penicilinas/química , Piperacilina/química , EsterilizaçãoRESUMO
Diabetic foot ulcer (DFU) is a common and devastating complication in diabetes. Antimicrobial resistance mediated by extended-spectrum ß-lactamases (ESBLs) production by bacteria is considered to be a major threat for foot amputation. The present study deals with the detection of Escherichia coli and the prevalence of bla(TEM), bla(SHV) and bla(OXA) genes directly from biopsy and swab of foot ulcers of diabetic patients. In total, 116 DFU patients were screened, of which 42 suffering with severe DFUs were selected for this study. Altogether 16 E. coli strains were successfully isolated from biopsy and/or swab samples of 15 (35.71%) patients. ESBL production was noted in 12 (75%) strains. Amplification of ß-lactamase genes by multiplex PCR showed the presence of bla(CTX-M) like genes in 10 strains, bla(TEM) and bla(OXA) in 9 strains each, and bla(SHV) in 8 of the total 16 strains of E. coli. Out of the ten antibiotics tested, E. coli strains were found to be resistant to ampicillin (75%), cefoxitin (56.25%), cefazolin (50%), meropenem (37.5%), cefoperazone (25%), cefepime (31.25%), ceftazidime (56.25%), and cefotaxime (68.75%) but all showed sensitivity (100%) to clindamycin and piperacillin-tazobactam. 3D models of the most prevalent variants of ß-lactamases namely TEM-1, SHV-1, OXA-1, and ESBL namely CTX-M-15 were predicted and docking was performed with clindamycin and piperacillin-tazobactam to reveal the molecular basis of drug sensitivity. Docking showed the best docking score with significant interactions, forming hydrogen bond, Van der Waals and polar level interaction with active site residues. Findings of the present study may provide useful insights for the development of new antibiotic drugs and may also prevent ESBLs-mediated resistance problem in DFU. The novel multiplex PCR assay designed in this study may be routinely used in clinical diagnostics of E. coli and associated bla(TEM), bla(SHV), and bla(OXA) like genes.
Assuntos
Antibacterianos/química , Clindamicina/química , Escherichia coli/efeitos dos fármacos , Ácido Penicilânico/análogos & derivados , beta-Lactamases/química , beta-Lactamas/química , Antibacterianos/farmacologia , Clindamicina/farmacologia , Pé Diabético/complicações , Pé Diabético/tratamento farmacológico , Pé Diabético/microbiologia , Escherichia coli/enzimologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/complicações , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/microbiologia , Expressão Gênica , Humanos , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Reação em Cadeia da Polimerase Multiplex , Ácido Penicilânico/química , Ácido Penicilânico/farmacologia , Piperacilina/química , Piperacilina/farmacologia , Combinação Piperacilina e Tazobactam , Homologia Estrutural de Proteína , Relação Estrutura-Atividade , Resistência beta-Lactâmica/efeitos dos fármacos , Inibidores de beta-Lactamases , beta-Lactamases/genética , beta-Lactamas/farmacologiaRESUMO
BACKGROUND: Several aerosol antibiotics are on the market and several others are currently being evaluated. Aim of the study was to evaluate the aerosol droplet size of five different antibiotics for future evaluation as an aerosol administration. MATERIALS AND METHODS: The nebulizers Sunmist(®), Maxineb(®) and Invacare(®) were used in combination with four different "small <6 ml" residual cups and two "large <10 ml" with different loadings 2-4-6-8 ml (8 ml only for large residual cups) with five different antibiotic drugs (ampicilln-sulbactam, meropenem, ceftazidime, cefepime and piperacillin-tazobactam). The Mastersizer 2000 (Malvern) was used to evaluate the produced droplet size from each combination RESULTS: Significant effect on the droplet size produced the different antibiotic (F=96.657, p<0.001) and the residual cup design (F=68.535, p<0.001) but not the different loading amount (p=0.127) and the nebulizer (p=0.715). Interactions effects were found significant only between antibiotic and residual cup (F=16.736, p<0.001). No second order interactions were found statistically significant. CONCLUSION: Our results firstly indicate us indirectly that the chemical formulation of the drug is the main factor affecting the produced droplet size and secondly but closely the residual cup design.
Assuntos
Antibacterianos/química , Nebulizadores e Vaporizadores , Aerossóis , Ampicilina/química , Cefepima , Ceftazidima/química , Cefalosporinas/química , Química Farmacêutica , Desenho de Equipamento , Meropeném , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/química , Piperacilina/química , Combinação Piperacilina e Tazobactam , Sulbactam/química , Tienamicinas/químicaRESUMO
PURPOSE: The physical compatibility of vancomycin and piperacillin sodium-tazobactam at dosing concentrations commonly administered during prolonged infusions was studied. METHODS: Concentrations of vancomycin and piperacillin sodium-tazobactam typically used in prolonged infusions were evaluated. Vancomycin hydrochloride and piperacillin sodium-tazobactam were reconstituted with 0.9% sodium chloride injection and diluted to the following concentrations: vancomycin, 4 mg/mL; piperacillin sodium 30 mg/mL plus tazobactam 3.75 mg/mL; and piperacillin sodium 40 mg/mL plus tazobactam 5 mg/mL. Combinations of vancomycin and piperacillin sodium-tazobactam were tested using simulated Y-site administration; phenytoin served as a positive control for precipitation with vancomycin. Each combination was prepared in triplicate, alternating the order of drug addition, and stored without light protection at room temperature. The resultant admixtures were microscopically observed and subjected to particle-size and turbidity analyses for five days. Statistical analyses were performed using two-way repeated measures analysis of variance with conservative Bonferroni corrections for multiple comparisons. RESULTS: Vancomycin was compatible with piperacillin sodium-tazobactam in the concentrations tested. No particulate matter was observed by the unaided eye. Similarly, the antibiotic admixtures displayed no differences microscopically, by particle-size analysis, or by turbidity analysis when compared with negative controls at five days. CONCLUSION: Vancomycin 4 mg/mL and piperacillin sodium 30 mg/mL plus tazobactam 3.75 mg/mL or piperacillin sodium 40 mg/mL plus tazobactam 5 mg/mL were physically compatible during simulated Y-site injection at room temperature without light protection for five days.