Targeting quorum sensing by designing azoline derivatives to inhibit the N-hexanoyl homoserine lactone-receptor CviR: Synthesis as well as biological and theoretical evaluations.

To counteract bacterial resistance, we investigated the interruption of quorum sensing mediated by non-classical bioisosteres of the N-hexanoyl homoserine lactone with an azoline core. For this purpose, a set of selected 2-substituted azolines was synthesized, establishing the basis for a new protocol to synthesize 2-amino imidazolines. The synthesized compounds were evaluated as inhibitors of violacein production in Chromobacterium violaceum. Theoretical studies on bioisostere-protein interactions were performed using CviR. The results show that some azolines decreased violacein production, suggesting an antiquorum sensing profile against Gram-negative bacteria. Docking and molecular dynamic simulations together with binding free energy calculations revealed the exact binding and inhibitory profiles. These theoretical results show relationship with the in vitro activity of the azoline series.

Synthesis, primary photophysical and antibacterial properties of naphthyl ester cinoxacin and nalidixic acid derivatives.

We have synthesized two naphthyl ester quinolone derivates and determined their ability to generate reactive oxygen species (ROS) such as (1)O(2), ()OH, H(2)O(2) upon photolysis with UV-A light. The ability of cinoxacin (1) and nalidixic acid (2), and their naphthyl ester derivatives (3 and 4) to generate a dose-dependent amount of singlet oxygen and ROS (()(-)O(2), ()OH) in cell-free systems was detected by histidine assay and by luminol-enhanced chemiluminescence (LCL), respectively. Their electronic absorption and emission spectra were quantified and their photostability was determined. Their tendency to generate peroxidic derivative species showed the following order: 3>4; in contrast, their ability to generate singlet oxygen was 4>3 and these were better sensitizers than their parent quinolones 1 and 2. The antibacterial activity in darkness and under irradiation of compounds 3 and 4 was tested on Escherichia coli and compared with that of their parent compounds. An enhanced antibacterial activity by irradiation of the naphthyl esters of cinoxacin and nalidixic acid on E. coli was observed.

Endosymbiotic alga from green hydra under the influence of cinoxacin.

Cinoxacin (Cxn) showed a strong effect on the endosymbiotic alga Chlorella; it was significantly damaged. Changes in algal color, position, structure and ultrastructure were found. In some algal cells ultrastructures were completely destroyed. The antichloroplastal and antimitochondrial effect was especially expressed. Damage to the thylakoid system of chloroplasts was more pronounced with increasing Cxn concentration. Some of the mitochondria were swollen and some of them were completely destroyed. From the evolutionary point of view, the correlation between antibacterial, and antichloroplastal and antimitochondrial effect of Cxn points to the evolutionary connection of chloroplasts and mitochondria with eubacteria.

Selective action of fluoroquinolones against intracellular amastigotes of Leishmania (Viannia) panamensis in vitro.

We have demonstrated that fluoroquinolones, a class of antibacterial agents that act through inhibition of type II DNA topoisomerases, exert selective action against intracellular amastigotes of Leishmania (Viannia) panamensis at concentrations that are achievable in vivo. Drug cytotoxicity assays employing the luciferase reporter gene revealed that intracellular amastigotes were 6.6- to 25.9-fold more sensitive than human macrophages (P < 0.05) to second-generation fluoroquinolones in vitro. The most selective agents (enoxacin and ciprofloxacin) exhibited 2 orders of magnitude greater potency against parasites (50% effective dose [ED50] = 54.9-83.4 microM) than host cells (ED50 = 1,425-1,740 microM). Linear regression analysis of ED50 data confirmed a complete lack of correlation (r = 0.001) between the relative drug sensitivities of parasites and host cells. A potential relationship between the structures of fluoroquinolones and their relative leishmanicidal activities was observed. The key substituents of the basic pyridone beta-carboxylic acid nucleus accounting for enhanced antiparasite potency and selectivity appear to be a nitrogen at position 8 of the bicyclic nucleus (enoxacin), a cyclopropyl substituent at the R1 site (ciprofloxacin), and linkage of the R1 and X8 groups by a CH3CHO bridge to form a tricyclic compound (ofloxacin). These findings support the potential of fluoroquinolones and derivatives as novel antileishmanials and encourage their clinical evaluation.

Corneal and scleral permeability of quinolones--a pharmacokinetics study.

PURPOSE: To investigate the corneal and scleral permeability of nalidixic acid and synthesized fluoroquinolones and their in vivo pharmacokinetics in rabbits. METHODS: The corneal and scleral permeability coefficients of ciprofloxacin, norfloxacin, cinoxacin, enoxacin, and ofloxacin were determined in rabbits using high performance liquid chromatography (HPLC). The aqueous humor levels of norfloxacin and ciprofloxacin were measured separately by topical instillation of 0.3% solutions of the two drugs onto rabbit eyes. RESULTS: Nalidixic acid had a higher corneal permeability coefficient (17.3 +/- 3.56 x 10(-6) cm/second) than all other drugs tested (p < 0.01). Corneal permeability coefficients in rabbits among ciprofloxacin, norfloxacin, cinoxacin, enoxacin, and ofloxacin were not significantly different (p > 0.1). Comparing the corneal and scleral permeability coefficients, only values for nalidixic acid were not significantly different (17.35 +/- 3.56 x l0(-6) cm/second versus 22.69 +/- 5.19 x 10(-6) cm/second, p > 0.05), while all other drugs had scleral permeability coefficients 8 to 10 times greater than corneal permeability coefficients. The mean aqueous humor concentration of norfloxacin and ciprofloxacin at 60 minutes to 180 minutes after instillation was around 0.3 microg/mL, a value higher than MIC90 of most bacteria.

Interactions of metal ions with two quinolone antimicrobial agents (cinoxacin and ciprofloxacin). Spectroscopic and X-ray structural characterization. Antibacterial studies.

Several novel metal-quinolone compounds have been synthesized and characterized by analytical, spectroscopic and X-ray diffraction methods. The crystal structure of the four compounds, Na(2)[(Cd(Cx)3)(Cd(Cx)3(H2O))].12H2O, [Co(Cp)2(H2O)2].9H2O, [Zn(Cp)2(H2O)2].8H2O and [Cd(HCp)2(Cl)2].4H2O, is presented and discussed: HCx=1-ethyl-1,4-dihydro-4-oxo(1,3)-dioxolo(4,5-g)cinnoline-3-carboxylic acid and HCp=1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3-quinoline carboxylic acid. In all these compounds the quinolone acts as a bidentate chelate ligand that binds through one carboxylate oxygen atom and the exocyclic carbonyl oxygen atom. Complexes of ciprofloxacin were screened for their activity against several bacteria, showing activity similar to that of the ligand. In addition, the number of bacteria killed after 3 h of incubation with the ligand, [Co(Cp)2(H2O)2].9H2O, Ni(Cp)2.10H2O and Cu(Cp)2.6H2O, was determined against S. aureus ATCC25923. There is a direct relationship between the growth rate and the lethal rate. Against growing bacteria, the ligand is the most bactericidal and Cu(Cp)2.6H2O is the less bactericidal. On the contrary, against non-dividing bacteria, the complexes were more bactericidal than the ligand, with Cu(Cp)(2).6H(2)O the most bactericidal compound.

Cinoxacin complexes with divalent metal ions. Spectroscopic characterization. Crystal structure of a new dinuclear Cd(II) complex having two chelate-bridging carboxylate groups. Antibacterial studies.

Several cinoxacin (HCx) complexes with divalent metal ions have been prepared and characterized by spectroscopic techniques. The crystal structure of [Cd2(Cx)4(H2O)2].10H2O has been determined by X-ray diffraction. The complex is triclinic, space group P1 with unit-cell dimensions: a = 10.412(2), b = 11.119(2), c = 13.143(6)A, chi== 76.78(4) degrees, beta = 74.59(3) degrees, gamma = 77.12(3) degrees, V = 1406.0(8) A3. In this complex each cadmium atom is heptacoordinated: the metal environment is formed by two Oketo and two Ocarbox atoms from two different cinoxacinate monoanions, two carboxylate oxygen atoms from a third cinoxacinate ligand and by one water oxygen atom on the seventh position. Two of the cinoxacinate ions act as tridentate chelate and bridging ligands and the other one as a bidentate chelate ligand. In the bridging monoanions the carboxylate group is behaving as a chelate ligand. All the complexes were screened for their activity against several bacteria, showing activity similar to that of cinoxacin. Additionally, the number of bacteria killed after 3 h of incubation with cinoxacin, [Cu(Cx)2].2H2O and [Co(Cx)3]Na.10H2O complexes was determined against E. coli ATCC 25922; the copper compound presents paradoxical effect which has been described and related to the mechanism of action of quinolones.

Synthesis, characterization, and crystal structure of [Cu(cinoxacinate)2].2H2O complex: a square-planar CuO4 chromophore. Antibacterial studies.

The structural and spectroscopic properties of a new copper (II) complex of cinoxacin (HCx) have been investigated. The complex [Cu(Cx)2].2H2O crystallizes in the monoclinic system, space group P2(1)/c. The cell dimensions are: a = 7.998(2), b = 7.622(1), c = 18.955(6) A, beta = 94.38(2) degree, V = 1154.6(6) A3, Z = 2. The structure was refined to R = 0.051. The crystal is composed of [Cu(Cx)2] units and uncoordinated water molecules. The Cu(II) ion, at a center of symmetry, is coordinated to two cinoxacinate (Cx) ligands related by the inversion center. Each cinoxacinate acts as bidentate ligand bonded to the cation through its carboxylate oxygen atom and through its exocyclic carbonyl oxygen atom, resulting in a CuO4 chromophore in a crystallographically planar configuration. The complex was screened for its activity against several bacteria, showing the same antimicrobial activity as the corresponding ligand.

Effect of nalidixic acid, pipemidic acid and cinoxacin on chondrocyte metabolism in explants of articular cartilage.

Explants of immature bovine articular cartilage were exposed to nalidixic acid, pipemidic acid and cinoxacin at one and ten times the human therapeutic plasma level for 7 days. Only nalidixic acid had significant effects on the chondrocyte metabolism. 20 micrograms/ml nalidixic acid caused an increase of 35S-sulfate incorporation into glycosaminoglycans at day 7. Two hundred micrograms/ml nalidixic acid inhibited the incorporation of 3H-thymidine into DNA. The incorporation of 35S-sulfate into glycosaminoglycans was decreased at day 0, while at day 7 the incorporation had returned to the control value. Pipemidic acid and cinoxacin had no significant effects on either the 3H-thymidine or the 35S-sulfate incorporation.

[Comparative evaluation of norfloxacin and cinoxacin in the therapy of complicated infections of the urinary tract]. / Valutazione comparativa della norfloxacina e della cinoxacina nella terapia delle infezioni complicate delle vie urinarie.

A randomized clinical study was conducted in order to compare the effectiveness of norfloxacin and cinoxacin in complicated urinary tract infections. Norfloxacin proved effective in 80% of the 40 patients treated, cinoxacin in only 60%.

Absence of cinoxacin-induced DNA fragmentation and mutations in the rat granuloma pouch.

The mutagenic and DNA-damaging activities of cinoxacin were evaluated in the rat granuloma pouch assay (GPA) in order to assess its genotoxic potential in vivo. High doses of this antimicrobial quinolone, either directly injected into the pouch or administered by gavage, did not induce mutation at the hgprt locus or DNA fragmentation in granuloma cells. Moreover, DNA damage was absent in the liver and kidney of rats given cinoxacin by the oral route.

Fluorocinnoline derivatives. II. Synthesis and antibacterial activity of fluorinated 1-alkyl-1,4-dihydro-4-oxocinnoline-3-carboxylic acids.

Chemical modification of cinoxacin was studied with the aim of improving its antibacterial activity and spectrum. Alkylation of ethyl 6,7,8-trifluoro- and 6,7-difluoro-4-hydroxycinnoline-3-carboxylates (1 and 7) with alkyl iodide or dialkyl sulfate gave ethyl 1-alkyl-6,7,8-trifluoro- and 6,7-difluoro-1,4-dihydro-4-oxocinnoline-3-carboxylates (2 and 8), together with the isomeric anhydro-bases 3 and 9 of 2-alkyl-3-ethoxycarbonyl-6,7,8-trifluoro- and 6,7-difluoro-4-hydroxycinnolinium hydroxides, respectively. Acid-catalyzed hydrolysis of the 1-alkyl derivatives 2 and 8 gave the corresponding carboxylic acids 4 and 10. The same treatment of 3 and 9, accompanied with decarboxylation of the inner salts 5 and 11, afforded the anhydro-bases 6 and 12 of 2-alkyl-4-hydroxycinnolinium hydroxides, respectively. Displacement reactions of 4 and 10 with nucleophiles such as amine, alkoxide and thiolate gave 7-substituted 1-alkyl-6,8-difluoro- and 6-fluoro-1,4-dihydro-4-oxocinnoline-3-carboxylic acids (13 and 17-35). Antibacterial activities of these compounds were evaluated and compared with those of cinoxacin and norfloxacin. Some compounds showed a broader spectrum and more potent activity than cinoxacin, but were considerably inferior in activity to norfloxacin.

Susceptibility of Bordetella pertussis to five quinolone antimicrobic drugs.

Five quinolone antimicrobic agents were tested to determine the mean inhibitory concentration (MIC) of each of 17 clinical strains of Bordetella pertussis by the agar dilution method. Ciprofloxacin demonstrated the best in vitro activity with an MIC90 of 0.06 microgram/ml. Norfloxacin, ofloxacin and enoxacin were also highly active with MIC90s of 0.25, 0.25, and 0.5, respectively. Cinoxacin was only moderately active (MIC90 4.0 microgram/ml).

Estudo clínico e microbiológico de cinoxacin na terapia de infecçöes do trato urinário / Clinical and microbiologic study of cinoxacin in the therapy of urinary tract infections

Trinta e dois pacientes com infecçäo urinária aguda inicial ou recorrente foram tratados com Cinoxacin na dose de 250 mg cada 12 horas por via oral. Vinte e sete pacientes (85%) apresentaram uma resposta clínica e bacteriológica satisfatória. Os pacientes näo apresentaram efeitos colaterais importantes

[Clinical study of cinoxacin in acute simple cystitis].

Cinoxacin (CINX) was administered twice a day for 7 consecutive days (400 mg X 2/day) to 34 female patients suffering from acute simple cystitis. The overall clinical efficacy was excellent in 15 cases (94%) and moderate in one case (6%) according to the criteria for clinical evaluation by the UTI committee. The efficacy was not determined in 18 cases. Bacteriological examination revealed 11 cases of single infection by E. coli, 2 cases of single infection by P. cepacia and S. epidermidis and one case of single infection by S. sunguis. MIC of E. coli ranged from 3.13 to 6.25 micrograms/ml. MIC of P. cepacia was 3.13 micrograms/ml and MIC of S. epidermidis more than 100 micrograms/ml. All the strains were eradicated with the efficacy of 100%. There was no relapse of acute simple cystitis in 16 cases after 7 days treatment of CINX. No serious side effects were recognized except for slight general fatigue and heart burn in 2 cases. It was thus concluded that CINX is clinically effective and safe for acute simple cystitis caused by E. coli and P. cepacia.

[In vitro comparative activity of norfloxacin in the comparison of pathogenic bacteria of the urinary tract]. / Attivita' comparativa in vitro di norfloxacina nei confronti di batteri patogeni del tratto urinario.

In this study 411 bacterial isolates from clinical urinary tract infections against some quinolones, azthreonam and trimethoprim + sulfamethoxazole combination were tested. Bactericidal kinetics of norfloxacin , cinoxacin and azthreonam against E. coli ATCC 25922 was determined. Both MICs values and bactericidal activities have shown that fluoroquinolones are more active against all Gram-positive and Gram-negative bacteria tested than the other drugs.

Frequency and expression of mutational resistance to the 4-quinolone antibacterials.

The frequency with which Escherichia coli mutated to resist a series of ten 4-quinolone antibacterials was studied. It was found that the mutation frequency could not be predicted from the potency of the drugs against sensitive bacteria. The mutation rates were lowest with ofloxacin, norfloxacin and ciprofloxacin. No mutants were observed with ofloxacin while less mutants were observed with norfloxacin than with ciprofloxacin, despite the latter possessing greatest activity against susceptible bacteria. The mutation frequencies observed with nalidixic acid, cinoxacin, pipemidic acid, flumequine, rosoxacin, oxolinic acid, and enoxacin were much higher than those observed with the three 4-quinolones mentioned earlier. When the sensitivities of the mutants to 4-quinolones were investigated, it was again found that the potency of each 4-quinolone against sensitive bacteria did not correlate with the levels of resistance of mutants to that drug. However, when the relative activities of the 4-quinolones against the mutants were investigated, it was found that no mutant isolated resisted a concentration exceeding the peak serum levels of ciprofloxacin, norfloxacin or ofloxacin. Although some mutants exhibited more resistance than the maximum attainable serum levels of the other 4-quinolones, the sensitivity of all the mutants fell well within the urine levels of pipemidic acid, flumequine, rosoxacin, oxolinic acid, enoxacin, norfloxacin, ofloxacin, and ciprofloxacin.