Effect of amoxicillin exposure on brain, gill, liver, and kidney of common carp (Cyprinus carpio): The role of amoxicilloic acid.

Amoxicillin (AMX) is one of the most commonly prescribed antibiotics around the world due to its broad-spectrum activity against different bacterial strains as well as its use as a growth promoter in animal husbandry. Although residues of this antibacterial agent have been found in water bodies in diverse countries, there is not enough information on its potential toxicity to aquatic organisms such as the common carp Cyprinus carpio. This study aimed to evaluate AMX-induced oxidative stress in brain, gill, liver and kidney of C. carpio. Carp were exposed to three different concentrations of AMX (10 ng/L, 10 µg/L, 10 mg/L) for 12, 24, 48, 72, and 96 h, and the following biomarkers were evaluated: lipid peroxidation (LPX), hydroperoxide content (HPC), protein carbonyl content (PCC) and activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Amoxicillin and its main degradation product amoxicilloic acid (AMA) were determined by high performance liquid chromatography coupled with electrochemical detection and UV detection (HPLC-EC-UV). Significant increases in LPX, HPC, and PCC (P < 0.05) were found in all study organs, particularly kidney, as well as significant changes in antioxidant enzymes activity. Amoxicilloic acid in water is concluded to induce oxidative stress in C. carpio, this damage being highest in kidney. The biomarkers used are effective for the assessment of the environmental impact of this agent on aquatic species. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1102-1120, 2017.

The influence of the carrier molecule on amoxicillin recognition by specific IgE in patients with immediate hypersensitivity reactions to betalactams.

The optimal recognition of penicillin determinants, including amoxicillin (AX), by specific IgE antibodies is widely believed to require covalent binding to a carrier molecule. The nature of the carrier and its contribution to the antigenic determinant is not well known. Here we aimed to evaluate the specific-IgE recognition of different AX-derived structures. We studied patients with immediate hypersensitivity reactions to AX, classified as selective or cross-reactors to penicillins. Competitive immunoassays were performed using AX itself, amoxicilloic acid, AX bound to butylamine (AXO-BA) or to human serum albumin (AXO-HSA) in the fluid phase, as inhibitors, and amoxicilloyl-poli-L-lysine (AXO-PLL) in the solid-phase. Two distinct patterns of AX recognition by IgE were found: Group A showed a higher recognition of AX itself and AX-modified components of low molecular weights, whilst Group B showed similar recognition of both unconjugated and conjugated AX. Amoxicilloic acid was poorly recognized in both groups, which reinforces the need for AX conjugation to a carrier for optimal recognition. Remarkably, IgE recognition in Group A (selective responders to AX) is influenced by the mode of binding and/or the nature of the carrier; whereas IgE in Group B (cross-responders to penicillins) recognizes AX independently of the nature of the carrier.

UNUSUALLY STABLE ADDUCT BETWEEN METHANOLYZED AMOXICILLIN OR AMPICILLIN AND THEIR DIKETOPIPERAZINE DERIVATIVES.

Amoxicillin and ampicillin were subjected to methanolysis. As expected, the methanolysis products were observed by HPLC-ESI-MS. Besides these products, diketopiperazine derivatives were also detected. Additionally, unusually stable adduct formed between the products of methanolysis and diketopiperazine derivatives was also identified. Analogical adducts were detected when ethanolysis was performed instead of methanolysis. HPLC-ESI-MS analysis of the separated adducts confirmed that the adducts were composed of methanolysis products and diketopiperazine derivatives.

Application of capillary electrophoresis to the simultaneous determination and stability study of four extensively used penicillin derivatives

The applicability of capillary electrophoresis for the analysis of four extensively used penicillin derivatives (benzylpenicillin, ampicillin, amoxicillin, oxacilllin) has been studied. Because of structural similarities, the electrophoretic behavior of these derivatives is very similar; consequently an efficient separation using the conventional capillary zone electrophoresis is hard to be achieved. Their simultaneous separation was solved by using micellar electrokinetic capillary chromatography, the separation being based on the differential partition of the analytes between the micellar and aqueous phase. Using a buffer solution containing 25 mM sodium tetraborate and 100 mM sodium dodecyl sulfate as surfactant, at a pH of 9.3, applying a voltage of + 25 kV at a temperature of 25 °C, we achieved the simultaneous separation of the studied penicillin derivatives in less then 5 minutes. The separation conditions were optimized and the analytical performance of the method was evaluated in terms of precision, linearity, limit of detection, and quantification. Also, a simple capillary zone electrophoresis method was applied to study the stability of the studied penicillin derivatives in water at different temperatures, using ciprofloxacin hydrochloride as internal standard. It was observed that the extent of the hydrolysis of penicillins in water is highly dependent on the time and also temperature.

Estudou-se a aplicabilidade de electroforese capilar para a análise de quatro derivados de penicilina (benzilpenicilina, ampicilina, amoxicilina, oxacilina) amplamente utilizados. Em razão das semelhanças estruturais, o comportamento electroforético destes derivados é muito semelhante e, por conseguinte, a separação eficaz utilizando a electroforese capilar de zona convencional é difícil de ser efetuada. A separação simultânea foi realizada por cromatografia capilar electrocinética micelar, que se baseia na partição diferencial entre os analitos na fase micelar e aquosa. Utilizando-se solução tampão contendo 25 mM de tetraborato de sódio e 100 mM de dodecil sulfato de sódio, como agente tensioativo, com pH de 9,3, voltagem de +25 kV, à temperatura de 25 °C, obteve-se a separação simultânea das penicilinas estudadas em menos de 5 minutos. As condições de separação foram otimizadas e o desempenho do método analítico foi avaliado em termos de precisão, linearidade, limite de detecção e de quantificação. Além disso, aplicou-se método de electroforese capilar de zona simples para estudar a estabilidade de penicilinas em água a diferentes temperaturas, utilizando cloridrato de ciprofloxacino como padrão interno. Estabeleceu-se que o grau de hidrólise de penicilinas em água é altamente dependente do tempo e também da temperatura.

Synthesis of 2-oxoquinoline-3-carboxamide of ampicillin and amoxicillin as inhibitors of penicillin binding protein 1A of pseudomonas aeruginosa.

A series of ampicillin and amoxicillin derivatives were prepared by N-acylation with N1-substituted 1, 2-dihydro-2-oxoquinoline carboxylic acids by Schotten-Baumann procedure and evaluated as inhibitors of Pseudomonas aeruginosa using molecular docking study. Most of the derivatives showed remarkableactivity against Gram positive and Gram negative bacteria in vitro.

Rapid and simultaneous determination of amoxicillin, penicillin G, and their major metabolites in bovine milk by ultra-high-performance liquid chromatography-tandem mass spectrometry.

A rapid, sensitive, and specific method for the determination of amoxicillin (AMO), amoxicilloic acid (AMA), amoxicillin diketopiperazine-2',5'-dione (DIKETO), penicillin G (PEN G), benzylpenicilloic acid (BPA-1), benzylpenilloic acid (BPA-2), and benzylpenillic acid (BPA-3) in bovine milk using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated. The method used penicillin V (PEN V) as the internal standard and ethanol for the deproteinisation of bovine milk. Chromatographic separation of the components was performed on a Waters Acquity UPLC® HSS T3 column (100 mm x 2.1 mm, 1.8 µm) using a mixture of 0.15% formic acid in water with 5mM ammonium acetate and acetonitrile as the mobile phase. Gradient elution was performed at a flow rate of 0.25 mL min⁻¹. The mass spectrometer was operated in the positive electrospray ionisation MS/MS mode. The method was fully validated according to EU requirements, including linearity, precision, trueness, limit of quantification, limit of detection, and specificity. The results were within the ranges specified. The established method was successfully applied in the determination of AMO, PEN G, and their major metabolites in 40 commercial bovine milk samples. The results showed that 8 samples were contaminated with BPA-1 or BPA-2. The mean levels (occurrence) of BPA-1 and BPA-2 in positive samples were 287 (50%) and 320 (100%) ng mL⁻¹, respectively. No sample was found to be contaminated with AMO, AMA, DIKETO, PEN G, and BPA-3. These findings could play an important role in food safety, because BPA-1 and BPA-2 metabolites pose possible health risks, although they are not included in the maximum residue limit legislation.

Tissue depletion of amoxicillin and its major metabolites in pigs: influence of the administration route and the simultaneous dosage of clavulanic acid.

A residue depletion study of amoxicillin (AMO) and its major metabolites, amoxicilloic acid (AMA) and amoxicillin diketopiperazine-2',5'-dione, was performed after a single oral (p.o.) and intravenous (i.v.) administration of amoxicillin (20 mg kg (-1)) and amoxicillin/clavulanic acid (20 and 5 mg kg (-1)) to pigs. Animals were slaughtered 12, 36, 48, 60, 72, and 84 h after dosing. Tissue samples were analyzed using liquid chromatography-tandem mass spectrometry. Kidney samples contained high concentrations of amoxicilloic acid metabolite, which depleted much slower from tissues than amoxicillin, both after p.o. (t1/2AMO = 4.5 h vs t1/2AMA = 8 h) and i.v. (t1/2AMO = 4 h vs t1/2AMA = 8 h) administration. Moreover, after oral administration, significantly higher amoxicilloic acid concentrations were measured in liver and kidney than after i.v. administration. The coadministration of amoxicillin with clavulanic acid provoked no significant differences in amoxicilloic acid tissue concentrations as compared to an amoxicillin dosing. The prolonged presence of residues of amoxicilloic acid in edible tissues can play an important role in food safety, because the compound could give rise to a possible health risk, although it is not included in the maximum residue limit legislation.

Design, synthesis, and biological evaluation of a series of beta-lactam-based prodrugs.

By use of pro-dual-drug concept the synthesis of 6-beta-[(R)-2-(clavaminio-9-N-yl)-2-(4-hydroxyphenylacetamido)]penicillanic acid (10), 6-beta-[(R)-2-(amino)-2-(4-(clavulano-9-O-yl)phenylacetamido)]penicillanic acid (13), (Z)-4-[2-(amoxycillin-4-O-yl)ethylidene]-2-(clavulano-9-O-yl)-3-methoxy-Delta(alpha,beta)-butenolide (19), and 3-[(amoxicillin-4-O-yl)methyl]-7-(phenoxyacetamido)-(1-oxo)-3-cephem-4-carboxylic acid (23) was accomplished. Unlike penicillin G, ampicillin, or amoxicillin, these four heretofore undescribed compounds 10, 13, 19, and 23 showed notable activity against beta-lactamase (betaL) producing microorganisms, Staphylococcus aureus A9606, S. aureus A15091, S. aureus A20309, S. aureus 95, Escherichia coli A9675, E. coli A21223, E. coli 27C7, Pseudomonas aeruginosa 18S-H, and Klebsiella pneumoniae A20634 TEM. In comparison with amoxicillin (9), alpha-amino-substituted compound 10 and butenolide derivative 19 showed a broadened spectrum of antibacterial activity; yet they were found to be less active than 13 and 23. Like clavulanic acid (7) or cephalosporin-1-oxide (21), the newly synthesized compounds 10, 13, 15, 16, 19, or 23 functioned as potent inhibitors of various bacterial betaLs.

Organometallic complexes with biological molecules. XVII. Triorganotin(IV) complexes with amoxicillin and ampicillin.

Novel triorganotin(IV) complexes of two beta-lactamic antibiotics, 6-[D-(-)-beta-amino-p-hydroxyphenyl-acetamido]penicillin (=amoxicillin) and 6-[D-(-)-alpha-aminobenzyl]penicillin (=ampicillin), have been synthesized and investigated both in solid and solution states. The complexes corresponded to the general formula R(3)Sn(IV)antib*H(2)O (R=Me, n-Bu, Ph; antib=amox=amoxicillinate or amp=ampicillinate). Structural investigations about configuration in the solid state have been carried out by interpreting experimental IR and 119Sn Mössbauer data. In particular, IR results suggested polymeric structures both for R(3)Sn(IV)amox.H(2)O and R(3)Sn(IV)amp*H(2)O. Moreover, both antibiotics appear to behave as monoanionic bidentate ligands coordinating the tin(IV) atom through ester-type carboxylate, as well as through the beta-lactamic carbonyl. Evidence that in none of these compounds water molecules were involved in coordination, was provided by thermogravimetric investigations. On the basis of 119Sn Mössbauer spectroscopy it can be inferred that tin(IV) was pentacoordinate in all of the complexes in the solid state, showing an equatorial R(3)Sn(IV) trigonal bipyramidal (tbp) configuration. The nature of the complexes in solution state was investigated by using 1H and 13C nuclear magnetic resonance (NMR) spectroscopy, while an 119Sn spectrum was obtained for n-Bu(3)Sn(IV)amp*H(2)O. Although 1H- and 13C-NMR measurements suggested that in dimethyl sulfoxide (DMSO)-d(6) solution the polymeric structure collapsed, due to a solvolysis process of the beta-lactamic carbonyl bonding to the organometallic moiety, the complexes have been shown to maintain the same trigonal bipyramidal configuration at tin(IV) atom by the coordination of a DMSO molecule. Cytotoxic activity of these novel semisynthetic antibiotic derivatives has been tested towards spermatocyte chromosomes of the mussel Brachidontes pharaonis (Mollusca: Bivalvia) using two different chromosome-staining techniques such as Giemsa and CMA(3). The occurrence of typical colchicinized-like (c-like) mitoses on slides obtained from animals exposed to organotin compounds, directly confirmed the high mitotic spindle-inhibiting potency of these chemicals. In addition, by comparative analysis of spermatocyte chromosomes from untreated specimens (negative controls) and specimens treated with the triorganotin(IV) complexes, structural damages such as 'achromatic lesions' and 'chromosome breakages' have been identified.

Antimicrobial susceptibility of Pasteurella multocida isolated from cattle and pigs.

Minimum inhibitory concentrations (MICs) of 10 antimicrobial agents were determined for Pasteurella multocida from cattle and pigs (72 and 68 isolates, respectively). Higher MICs were observed with oxytetracycline, doxycycline, tilmicosin and thiamphenicol for porcine isolates than for bovine isolates. Enrofloxacin was the most active, with an MIC for 90% of the isolates (MIC90) of 0.05 microg/ml for both bovine and porcine isolates. Aspoxicillin exhibited the same excellent activity against penicillin-susceptible isolates as ceftiofur, with MICs ranging from < or = 0.025 to 0.1 microg/ml. Aminoglycosides were less active, with an MIC90 of > 100 microg/ml for both bovine and porcine isolates.

Synthesis and antimicrobial activity of new pyrrolecarboxylic acid derivatives of ampicillin and amoxicillin.

Twelve new penicillin derivatives were prepared for microbiological evaluation by N-acylation of ampicillin (CAS 69-53-4) and amoxicillin (CAS 26787-78-0) with activated pyrrolecarbocylic acids via mixed anhydrides following the Schotten-Baumann procedure. An alternative synthetic approach via chloroanhydrides was checked and rejected because of the instability established for these intermediates. NMR and IR spectral data together with TLC confirmed the structure and the purity of the new products. Antimicrobial tests in vitro indicated a reduction of the antibacterial activity compared with that of ampicillin and amoxicillin as reference antibiotics, but their minimal inhibitory concentrations (MIC) were still in the range of 0.62-16 micrograms/ml against standard and clinical Gram positive strains. Preliminary toxicological evaluations showed low toxicity.

Beta-lactam antibiotics functionalized with gem-bisphosphonates.

Acylation of amoxycillin and cephalexin with acids III, V and VII, and with isocyanate VIII furnished the corresponding beta-lactam antibiotics (X and XIII-XV, respectively). The antibacterial activity of these new antibiotic analogues against Helicobacter pylori was found to be identical with those of amoxycillin, Augmentin, erythromycin and ciprofloxacin.

[Positive seroconversion syphilis in a patient with acute lymphocytic leukemia after allogeneic bone marrow transplantation].

An acute lymphocytic leukemia patient underwent allogeneic bone marrow transplantation (BMT) from a sibling who was serologically positive for syphilis. After the donor was administered antibiotic therapy, the titration of treponema pallidum hemagglutination (TPHA) decreased from x1260 to x320. Thereafter, the graft consisting of mononuclear cells was transplanted. TPHA of the recipient turned positive on day +63, but became negative 1.5 years after BMT. Although the cause of the seroconversion of TPHA seemed to be the contamination of treponema to the graft, the adoptive transfer could not be ruled out as an another possible cause.

[A clinical study on chemotherapies for chorioamnionitis].

Effects of imipenem/cilastatin (IPM/CS) therapy, flomoxef (FMOX) therapy and combined ceftazidime + aspoxicillin (CAZ/ASPC) therapy as initial therapies for chorioamnionitis were assessed clinically. 1. The subjects were 49 women with threatened abortion and 29 with premature rupture of membranes (PROM), complicated in all cases by chorioamnionitis. The inflammation was treated with IPM/CS in 19 patients, FMOX in 39, CAZ in 11, and CAZ/ASPC in 9. 2. The response rate to therapy for chorioamnionitis was 95.9% (47/49) in the threatened abortion group. Of the 49 patients in this group, 16 (32.7%) underwent premature labor. Of the therapies administered, IPM/CS tended to prevent premature labor more frequently than did any other therapy. The latent period (from rupture of membranes to delivery) was equal to or longer than 7 days in the PROM group. The percent prolongation of the latent period in these patients (55.5%) was significantly greater than that previously obtained with penicillin therapy. 3. The bacterial elimination rate was 50.9% (29/57). Of the 36 bacterial isolates, 66.7% were Gram-positive bacteria. The bacteriological efficacy rate was 89.7% (26/29). These results suggest that antibacterial agents effective against Gram-positive bacteria should be selected for treatment of chorioamnionitis, and that IPM/CS therapy is particularly useful considering the drug's good transfer into amniotic fluid and its antibacterial spectrum.

Aspoxicillin versus piperacillin in severe abdominal infections--a comparative phase III study.

We compared aspoxicillin, a new broad-spectrum penicillin derivative, with piperacillin in severe abdominal infection. Aspoxicillin 4 g administered tds (n = 52) or piperacillin 4 g qds (n = 53) usually as monotherapy were randomly given to patients suffering from perforated appendicitis, acute cholecystitis, ulcer or colon perforation, or intra-abdominal abscess. Blood, tissue and exudate cultures were obtained when applicable for pathogen identification and susceptibility testing. The efficacy rates were similar in the two study groups. Of the 50 evaluable aspoxicillin patients 45 (90%) were considered as treatment responders compared with 48 patients out of 53 (91%) in the piperacillin group (NS). The 95% confidence interval for the efficacy difference was -12% to +11% thus showing no difference between the two drugs. Both drugs were generally well tolerated and no serious drug-related adverse events were noted. However, five patients died because of their illness and one patient had a fatal myocardial infarction. In conclusion, aspoxicillin 4 g tds was shown to be equal to piperacillin 4 g qds in severe abdominal infections.

[A clinical study on the efficacy of ceftazidime and aspoxicillin in chorioamnionitis. Abdominal Infections Research Group].

Chorioamnionitis as a complication of threatened abortion and preterm labor and preterm PROM were treated with ceftazidime (CAZ) and aspoxicillin (ASPC) as a primary therapy. The following results were obtained. 1. Cases of threatened abortion and preterm labor (n = 25) and preterm PROM (n = 5) were treated with 2-4 g CAZ/day alone (n = 14) or in combination with 4 g ASPC/day (n = 16) along with a uterine contraction inhibitor (ritodrine hydrochloride etc. n = 28) and clinical evaluation was made. 2. In the cases of threatened abortion and preterm labor the efficacy ratio was 24/25 (96%). In the cases of preterm PROM, the latent period-delaying effect was observed in five out of the five patients. Upon analysis of the tocolysis index in the group of threatened abortion and preterm labor, the index values > or = 5 were observed in 12 out of 25 (60%), and the delivery incidence before the 35th week of gestation was 4/25 (16%). In all patients, the incidence of delivery after the 36th week of gestation was 24/30 (80%). 3. Bacteriological examinations showed a high detection rate for Gram-positive bacteria, and the combination effect between ASPC and CAZ was demonstrated against all 9 isolates examined. The above results indicated a high efficacy as well as safety of the combination of CAZ and ASPC as a primary therapeutic means against chorioamnionitis.

[Studies on aspoxicillin for its transfer to the skin].

To evaluate the usefulness of aspoxicillin (ASPC) in the field of plastic and reconstructive surgery, we examined its transfer to the skin. 1. After intravenous drip infusion of ASPC for 1 hour at a dose of 2 g in 13 adults and at 1 g in 2 children, the mean serum ASPC concentration 1 hour after termination of the infusion was 70.46 +/- 28.05 micrograms/ml. The mean concentration in the skin tissue 1 hour after infusion in 15 patients was 32.45 +/- 18.47 micrograms/g. The rate of transfer to the skin 1 hour after infusion in the 15 patients was 52.9 +/- 29.7%. 2. The ASPC concentrations in skin tissues and the rates of its transfer to the skin did not differ significantly between 5 patients with facial surgery and 10 with surgery in the trunk or limbs. 3. To prevent postoperative infections, ASPC was intravenously drip infused twice daily for 2 approximately 3 days after operation at a dose of 2 g in adults and 1 g in children. No postoperative infection occurred in any patient, suggesting the effectiveness of this drug. In addition, no side effects or abnormalities in clinical examination values were observed.

[A clinical study on the efficacy of ceftazidime alone or in combination with aspoxicillin against infections in obstetric and gynecological patients].

We studied clinical effects of ceftazidime (CAZ) alone or in combination with aspoxicillin (ASPC) against various infections in obstetric and gynecological patients. 1. Obstetric and gynecological patients (n = 91) with various infectious diseases were treated with CAZ alone (1-2 g x 2/day, n = 54) or in combination with ASPC (1-2 g x 2/day, n = 37) administered via drip infusion. 2. CAZ alone or in combination with ASPC was efficacious in 50 out of 54 (92.6%) or 33 out of 36 (91.7%) patients, respectively. Overall, the efficacy ratios were 46/49 (93.9%) against gynecological infections, 21/25 (84.0%) against perinatal infections and 16/16 (100%) against other infections. The bacteriological efficacy ratio was 21/21 (100%) while clinical effectiveness in cases in which causative agents were known was observed in 20 out of 21 (95.2%) patients. In patients who had not respond to other treatments, CAZ alone, and in combination with ASPC were effective in 15 out of 16 (93.8%) and 6 out of 8 (75.0%) patients, respectively, hence the overall efficacy ratio was 21/24 (87.5%). 3. Abnormal values in clinical laboratory tests were obtained in 3 out of 91 (3.3%) patients. No other adverse side effects were observed in any of the patients.