Anti-MRSA drug discovery by ligand-based virtual screening and biological evaluation.

S. aureus resistant to methicillin (MRSA) is one of the most-concerned multidrug resistant bacteria, due to its role in life-threatening infections. There is an urgent need to develop new antibiotics against MRSA. In this study, we firstly compiled a data set of 2,3-diaminoquinoxalines by chemical synthesis and antibacterial screening against S. aureus, and then performed cheminformatics modeling and virtual screening. The compound with the Specs ID of AG-205/33156020 was discovered as a new antibacterial agent, and was further identified as a Gyrase B (GyrB) inhibitor. In light of the common features, we hypothesized that the 6c as the representative of 2,3-diaminoquinoxalines also inhibited GyrB and eventually proved it. Via molecular docking and molecular dynamics simulations, we identified binding modes of AG-205/33156020 and 6c to the ATPase domain of GyrB. Importantly, these GyrB inhibitors inhibited the MRSA strains and showed selectivity to HepG2 and HUVEC. Taken together, this research work provides an effective ligand-based computational workflow for scaffold hopping in anti-MRSA drug discovery, and discovers two new GyrB inhibitors that are worthy of further development.

Determination of the Environmental Pollution Potential of Some Herbicides by the Assessment of Cytotoxic and Genotoxic Effects on Allium cepa.

The present study aims to evaluate the potential for the pollution of the environment by two herbicides (quizalofop-p-ethyl and cycloxydim), using the Allium test. The species in question is Allium cepa (onion, 2n = 16), one of the most common plant indicators of environmental pollution. The working method consisted of obtaining the meristematic roots of Allium cepa and their treatment with herbicides at three different concentrations (0.5%, 1%, and 1.5%) for each herbicide for 24 h, for comparison with an untreated control. The results obtained from the cytological study indicated a strong cytotoxic and genotoxic effect for both herbicides, but especially for quizalofop-p-ethyl, where the mitotic index decreased from 30.2% (control) to 9.6% for the variant treated with 1.5% herbicide. In this case, a strong mitodepressive effect was shown by a highly significant percentage (35.4%) of chromosomal aberrations and nuclear alterations: stickiness, fragments, C-mitosis, lobulated nucleus, micronuclei, and nuclear erosion. The mitodepressive effect as well as the percentage of chromosomal aberrations increased with a higher herbicide concentration. The obtained results suggest the strong potential for pollution of the two herbicides, particularly at concentrations higher than 0.5%; therefore, we recommend caution in their use to avoid undesirable effects on the environment.

Hepatoprotective effects of a Chinese herbal formulation, Yingchen decoction, on olaquindox-induced hepatopancreas injury in Jian carp (Cyprinus carpio var. Jian).

In order to identify effective hepatoprotective herbs for clinical application in fish farming, 200 mg/kg olaquindox (OLA) was added to a basal diet (group 1, control) to form OLA diet (group 2), then 1.35, 2.7 and 5.4 % (w/w) of a Chinese herbal formulation, Yingchen decoction (YCD), were added to the OLA diet to form three additional diets for groups 3, 4 and 5, respectively. A total of 375 juvenile Jian carp (Cyprinus carpio var. Jian) (52.12 ± 2.95 g/tail) were divided into five groups (triplicates per group) and fed the five diets mentioned above, respectively, for 6 weeks. At the termination of feeding experiment, serum biochemical indexes, viability of hepatocytes and the hepatopancreas microstructure for each group were detected and observed. The results showed that serum ALT and AST in group 2 were significantly higher than the control (P < 0.05). Plasma membranes hepatocyte nuclei in group 2 were found to be mostly indistinct, compared to group 1, and gradually recovered with the increasing supplementation of YCD in group 3, 4 and 5. The viability of isolated hepatocytes in group 2 was the lowest and gradually recovered with the increasing supplementation of YCD in group 3, 4 and 5. The results suggest that YCD protected the Jian carp hepatopancreas against injury from OLA, and that 5.4 % YCD would be the optimum dosage in a Jian carp diet.

Formation and mitigation of heterocyclic aromatic amines in fried pork.

Heterocyclic aromatic amines (HAAs) are potent mutagens and carcinogens generated during the heat processing of meat. HAAs, which are abundant in processed meat products, include 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), and 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP). The content of these three HAAs in fried pork was determined by LC-MS/MS. The effects of frying time and temperature, sample shape, and addition of antioxidants on the generation of HAAs were investigated. The results show that HAAs were produced during frying, and their levels increased with increasing frying time and temperature. Pork patties had the highest concentration of HAAs compared with pork meatballs and pork strips. The addition of antioxidant of bamboo leaves (AOB), liquorice extract, tea polyphenol, phytic acid and sodium iso-ascorbate to pork before frying had an inhibitory effect on HAA generation, with AOB being the most effective antioxidant. Inhibition levels of nearly 69.73% for MeIQx, 53.59% for 4,8-DiMeIQx and 77.07% for PhIP in fried pork were achieved when the concentrations of AOB added were 0.02, 0.01 and 0.10 g kg⁻¹, respectively.

Evaluation of extended release brimonidine intravitreal device in normotensive rabbit eyes.

PURPOSE: To evaluate the safety profile of a brimonidine extended release intravitreal implant, in normotensive rabbit eyes. METHODS: Devices were made from hollow poly-l-lactic acid (PLA) tubes and contained hundred micrograms of brimonidine pamoate. Device was injected intravitreally in one eye of 12 New Zealand pigmented rabbits, whereas other eye was injected with a sham implant in masked fashion. Ocular examination was conducted at baseline and months 1, 3 and 6 including dilated fundus examination and electro-retinogram (ERG). Four rabbits were sacrificed at each time-point for retinal histology. ERG data were compared between groups and time-points using anova. RESULTS: No complications were reported from either eye of any rabbits over a 6-month period. Photopic A wave was reduced in the control eye at 1 month compared with baseline (p < 0.01). There was no significant difference in other ERG parameters between the groups at different time-points. Gross retinal histology was normal at all time-points. CONCLUSION: Extended release intravitreal brimonidine device was found to be safe and in normotensive rabbit eyes.

Identification of thioredoxin glutathione reductase inhibitors that kill cestode and trematode parasites.

Parasitic flatworms are responsible for serious infectious diseases that affect humans as well as livestock animals in vast regions of the world. Yet, the drug armamentarium available for treatment of these infections is limited: praziquantel is the single drug currently available for 200 million people infected with Schistosoma spp. and there is justified concern about emergence of drug resistance. Thioredoxin glutathione reductase (TGR) is an essential core enzyme for redox homeostasis in flatworm parasites. In this work, we searched for flatworm TGR inhibitors testing compounds belonging to various families known to inhibit thioredoxin reductase or TGR and also additional electrophilic compounds. Several furoxans and one thiadiazole potently inhibited TGRs from both classes of parasitic flatworms: cestoda (tapeworms) and trematoda (flukes), while several benzofuroxans and a quinoxaline moderately inhibited TGRs. Remarkably, five active compounds from diverse families possessed a phenylsulfonyl group, strongly suggesting that this moiety is a new pharmacophore. The most active inhibitors were further characterized and displayed slow and nearly irreversible binding to TGR. These compounds efficiently killed Echinococcus granulosus larval worms and Fasciola hepatica newly excysted juveniles in vitro at a 20 µM concentration. Our results support the concept that the redox metabolism of flatworm parasites is precarious and particularly susceptible to destabilization, show that furoxans can be used to target both flukes and tapeworms, and identified phenylsulfonyl as a new drug-hit moiety for both classes of flatworm parasites.

Soil application of dinitroaniline and arylphenoxy propionic herbicides influences the activities of phosphate-solubilizing microorganisms in soil.

An experiment was conducted under laboratory conditions to investigate the effect of two systemic herbicides, viz. pendimethalin (a dinitroaniline) and quizalofop (an arylphenoxy propionic acid) at their recommended field application rates (1.0 kg and 50 g active ingredient per hectare, respectively), either separately or in a combination, on growth and activities of phosphate-solubilizing microorganisms in relation to their effects on biochemical transformations and availability of organic carbon, total and available phosphorus in a Typic Haplustept soil of West Bengal, India. Application of herbicides, in general, significantly stimulated the growth and activities of phosphate-solubilizing microorganisms which increased microbial biomass resulting in higher accumulation of oxidizable organic carbon, total and available phosphorus in soil as compared to untreated control. The combined application of both the herbicides highly stimulated the proliferations of phosphate-solubilizing microorganisms, while pendimethalin alone significantly accentuated phosphate-solubilizing capacities 36.4% as compared to untreated control and retained highest amount of total phosphorus due to greater microbial activities in soil. The separate application of quizalofop also manifested an induced effect on the proliferations of phosphate-solubilizing microorganisms and accounted significant amounts of organic carbon and available phosphorus in the soil system. The results of the present study thus indicated that the cited herbicides at their field application rates can be safely used to eradicate weeds in the crop fields.

Pu-erh black tea extract supplementation attenuates the oxidative DNA damage and oxidative stress in Sprague-Dawley rats with renal dysfunction induced by subchronic 3-methyl-2-quinoxalin benzenevinylketo-1,4-dioxide exposure.

3-Methyl-2-quinoxalin benzenevinylketo-1,4-dioxide (Quinocetone, QCT), has been used to treat dysentery and promote growth in animal feeding. However, available data show that QCT has potential nephrotoxicity. The present study was designed to investigate the protective effects of Pu-erh black tea extract (PBTE) which is a traditional remedy in China with antioxidant properties against oxidative DNA damage and oxidative stress in a rat model of QCT-induced renal dysfunction. Increased serum creatinine, blood urea nitrogen, pathological lesions, urinary 8-hydroxy 2-deoxyguanosine (8-OHdG) and renal DNA damage were observed in the QCT-fed rats. These were accompanied by intracellular reactive oxygen species accumulation, enhanced lipid peroxidation, and inhibited antioxidant system, i.e., glutathione glutathione S-transferase, glutathione peroxidase and glutathione reductase. Oral administration of PBTE effectively suppressed QCT-induced renal dysfunction, as evidenced by reduced serum creatinine, urinary 8-OHdG and DNA damage in isolated renal cells, amelioration of oxidative stress and modulation of antioxidative system. In conclusion, PBTE administration ameliorated QCT-induced nephrotoxicity by maintaining DNA's double-helix architecture and mitigating oxidative stress.

Pu-erh black tea supplementation decreases quinocetone-induced ROS generation and oxidative DNA damage in Balb/c mice.

Quinocetone (3-methyl-2-quinoxalinbenzenevinylketo-1,4-dioxide, QCT), a new feed antibacterial agent of quinoxaline-1,4-dioxides family, has been used as an animal growth promoter. However, few data about its potential toxicity in vivo were available. In this study, genotoxicity of QCT and the relationship with oxidative stress were investigated. Balb/c mice with both sexes were administrated with QCT (12000, 6000 and 3000 mg/kg/bw, respectively) by gavage acutely. DNA damage, generation of reactive oxygen species (ROS) and activity of antioxidative system (total antioxidative capacity, glutathione, glutathione peroxidase, superoxide dismutase and catalase) in liver and kidney were determined. Moreover, Pu-erh black tea extract (BTE) was co-administrated with QCT to evaluate its protective effect against QCT-induced genotoxicity. The DNA damage was observed in all the groups treated with single QCT except the liver with dose of 3000 mg/kg/bw. ROS was accumulated and antioxidative system was suppressed both in liver and kidney. However, the DNA damage, as well as the ROS, was decreased, while the activity of antioxidative system was increased in mice after co-administration of QCT and BTE. These data demonstrate that oxidative stress mediated the genotoxicity induced by QCT in vivo. Furthermore, this oxidative DNA damage can be attenuated by pre-supplementation of BTE.

[Genoprotective effect of sodium humate in conditions of induced oxidative stress].

It was shown that sodium humate in a range of concentration of 50-1000 mg/l reduces a level of clastogenic effects induced with dioxidin (20 mg/l), a prooxidant mutagene. The greatest effect was observed at concentration of 300 mg/l. Direct doze-effect dependence under the influence of sodium humate was not revealed. Possible antioxydative and desmutagenic mechanisms of sodium humate protective action are discussed.

[Mutagenesis induced with dioxidin in the Allium-test].

The influence of dioxidin in different concentrations (10-100 mg/l) on the cytogenetic parameters of Allium cepa L. has been studied. The mutagenic effect of dioxidin was shown within all the range of the studied concentrations. The curve "dose-effect" has been determined for the concentrations of 10, 20, 30, 40, 50, and 100 mg/l. The peak of the mutagenic effect and significant reduction of the mitotic index were revealed at the concentration of 100 mg/l. It was shown that the mutagenic efficiency of the dioxidin statistically correlated with reduction of the mitotic activities.

Phytoalexin resveratrol attenuates the mutagenicity of the heterocyclic amines 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline.

Resveratrol is a phytoalexin, that belongs to a family of naturally occurring stilbenes. It has been reported that resveratrol can inhibit chemical carcinogenesis in experimental animals and although the mechanisms involved are unknown, an anti-mutagen mechanism has been proposed. We have explored this hypothesis using mutagenicity assays based on bacterial (Salmonella typhimurium) and eukaryotic cells (Chinese hamster V79 cells). We found resveratrol to be potent in both systems, blocking the mutagenicity of the food-derived heterocyclic amines (HA) 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) at micromolar concentrations. Furthermore, in cells capable of activating 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine to cytotoxic derivatives, resveratrol was able to attenuate cytotoxicity. Paradoxically, in cells lacking the ability to activate PhIP, resveratrol itself was toxic and co-incubation with PhIP reduced this toxicity. Our data confirm the potent anti-mutagenic activity of resveratrol and support its potential as a chemopreventative.

Potent antimutagenic activity of white tea in comparison with green tea in the Salmonella assay.

There is growing interest in the potential health benefits of tea, including the antimutagenic properties. Four varieties of white tea, which represent the least processed form of tea, were shown to have marked antimutagenic activity in the Salmonella assay, particularly in the presence of S9. The most active of these teas, Exotica China white tea, was significantly more effective than Premium green tea (Dragonwell special grade) against 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and four other heterocyclic amine mutagens, namely 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). Mechanism studies were performed using rat liver S9 in assays for methoxyresorufin O-demethylase (MROD), a marker for the enzyme cytochrome P4501A2 that activates heterocyclic amines, as well as Salmonella assays with the direct-acting mutagen 2-hydroxyamino-3-methylimidazo[4,5-f]quinoline (N-hydroxy-IQ). White tea at low concentrations in the assay inhibited MROD activity, and attenuated the mutagenic activity of N-hydroxy-IQ in the absence of S9. Nine of the major constituents found in green tea also were detected in white tea, including high levels of epigallocatechin-3-gallate (EGCG) and several other polyphenols. When these major constituents were mixed to produce "artificial" teas, according to their relative levels in white and green teas, the complete tea exhibited higher antimutagenic potency compared with the corresponding artificial tea. The results suggest that the greater inhibitory potency of white versus green tea in the Salmonella assay might be related to the relative levels of the nine major constituents, perhaps acting synergistically with other (minor) constituents, to inhibit mutagen activation as well as "scavenging" the reactive intermediate(s).

Metabolic, idiosyncratic toxicity of drugs: overview of the hepatic toxicity induced by the anxiolytic, panadiplon.

Preclinical drug safety evaluation studies, typically conducted in two or more animal species, reveal and define dose-dependent toxicities and undesirable effects related to pharmacological mechanism of action. Idiosyncratic toxic responses are often not detected during this phase in development due to their relative rarity in incidence and differences in species sensitivity. This paper reviews and discusses the metabolic idiosyncratic toxicity and species differences observed for the experimental non-benzodiazepine anxiolytic, panadiplon. This compound produced evidence of hepatic toxicity in Phase 1 clinical trial volunteers that was not predicted by rat, dog or monkey preclinical studies. However, subsequent studies in Dutch-belted rabbits revealed a hepatic toxic syndrome consistent with a Reye's Syndrome-like idiosyncratic response. Investigations into the mechanism of toxicity using rabbits and cultured hepatocytes from several species, including human, provided a sketch of the complex pathway required to produce hepatic injury. This pathway includes drug metabolism to a carboxylic acid metabolite (cyclopropane carboxylic acid), inhibition of mitochondrial fatty acid beta-oxidation, and effects on intermediary metabolism including depletion of glycogen and disruption of glucose homeostasis. We also provide evidence suggesting that the carboxylic acid metabolite decreases the availability of liver CoA and carnitine secondary to the formation of unusual acyl derivatives. Hepatic toxicity could be ameliorated by administration of carnitine, and to a lesser extent by pantothenate. These hepatocellular pathway defects, though not directly resulting in cell death, rendered hepatocytes sensitive to secondary stress, which subsequently produced apoptosis and hepatocellular necrosis. Not all rabbits showed evidence of hepatic toxicity, suggesting that individual or species differences in any step along this pathway may account for idiosyncratic responses. These differences may be roughly applied to other metabolic idiosyncratic hepatotoxic responses and include variations in drug metabolism, effects on mitochondrial function, nutritional status, and health or underlying disease.

[Effect of plant melanin pigment on the clastogenic effects of chemical mutagens in mice]. / Vliianie rastitel'nogo melaninovogo pigmenta na klastogennye éffekty khimicheskikh mutagenov u myshei.

The chromosome aberration assay in the bone marrow cells of C57BL/6 mice showed that melanin pigment (MP) in a dose range from 0.01 to 10 mg/kg does not influence the clastogenic effect of dioxidine (200 mg/kg, i.p.), while reducing the clastogenic effect of cyclophosphamide (20 mg/kg, i.p.) by a factor of 1.5-4 in various treatment regimes depending on the mutagen injection time.

[Modification by lacidipine of the clastogenic effect of dioxidine in vivo]. / Modifikatsiia latsidipinom klastogennogo éffekta dioksidina in vivo.

The influence of lacidipine (0.1-10 mg/kg, intragastric) on the clastogenic effect of dioxidine (100 and 200 mg/kg, i.p.) under conditions of their single and repeated (five-fold, 24-h interval) administration was studied by the chromosome aberration assay in the metaphase bone marrow cells of BALB/c and C57BL/6 male mice. It was found that single (5 or 10 mg/kg) and repeated (10 mg/kg) introduction of lacidipine enhances the clastogenic effect of dioxidine in both genotypes. At the same time, a single treatment of C57BL/6 mice with 0.1 and 1 mg/kg of lacidipine sometimes significantly reduced the clastogenic effect of dioxidine (200 mg/kg). Thus, lacidipine exhibits a comutagen effect in vivo when administered at large doses (5 and 10 mg/kg) but not at small doses, where the drug sometimes acted as antimutagen in C57BL/6 mice.

Enhancing effects of Thai edible plants on 2-amino-3, 8-dimethylimidazo(4,5-f)quinoxaline-hepatocarcinogenesis in a rat medium-term bioassay.

Boesenbergia pandurata (Zingiberaceae), Languas galanga (Zingiberaceae) and Citrus hystrix (Rutaceae) are edible plants that are commonly used as flavors or condiments in various Thai food dishes. They are known to exert strong anti-promoting activity in a test of tumor promoter-induced Epstein-Barr virus (EBV) activation. In the present study their effects on hepatocarcinogenesis were investigated in a medium-term bioassay using F344 male rats. C. hystrix significantly enhanced 2-amino-3,8-dimethylimidazo(4, 5-f)quinoxaline-associated preneoplastic liver cell focus development while B. pandurata and L. galanga had borderline effects. The results suggest that C. hystrix as well as B. pandurata and L. galanga may contain agents augmenting the hepatocarcinogenicity of 2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline.

Effects of chronic administration of low doses of 2-amino-3,8-dimethylimidazo-[4,5-f]quinoxaline on glutathione S-transferase placental form-positive foci development in the livers of rats fed a choline-deficient diet.

Effects of chronic administration of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) at the very low doses of 0.4 and 4 ppm, respectively 1000- and 100-fold less than the dose shown to be carcinogenic (400 ppm), on the liver of rats fed a choline-deficient (CD) diet were examined in terms of glutathione S-transferase placental form (GST-P)-positive foci. Male F344 rats were given CD diet containing 0, 0.4 or 4 ppm MeIQx for 20 or 40 weeks. As controls, rats received choline-supplemented (CS) diet in the same manner. MeIQx at 4 ppm in the CD diet significantly increased both the number and area of GST-P-positive foci, the values being 2.3- and 2.1-fold at 20 weeks and 2.0- and 3.3-fold at 40 weeks, respectively, compared with those observed for CD diet alone. MeIQx at 0.4 ppm in CD diet did not affect the development of GST-P-positive foci. No influence of the heterocyclic amine was found in the CS groups, where only very small numbers of minute lesions were observed. The level of MeIQx-DNA adducts in rats given the CD diet containing 4 ppm MeIQx was 2- to 3-fold lower than that in rats given the CS diet containing 4 ppm MeIQx at 20 and 40 weeks. This result indicates that DNA adduct formation and cell proliferation are both required for the increase of GST-P-positive foci in rats fed 4 ppm MeIQx in a CD diet. The above findings strongly suggest that MeIQx could be carcinogenic even at 4 ppm under CD conditions, where liver cell regeneration is continuously occurring.

[Validation of the choice of antibacterial agents for treating periodontitis]. / Obosnovanie vybora antibakterial'nykh sredstv dlia lecheniia periodontita.

Sensitivity of microflora sampled from the dental root channels of periodontitis patients to chlorhexidine, dioxidine, quinosol, Metrodgile was investigated. Chlorhexidine and dioxidine were shown to act predominantly on gram-positive and gram-negative bacteria, Metrodgile most active against anaerobic flora. High antibacterial activity of quinosol and chlorhexidine was combined with their pronounced cytotoxic effects. Metrodgile could be successfully used for preventing purulent odontogenic inflammations caused by anaerobic flora.

Antiherpesvirus activity and mechanism of action of indolo-(2,3-b)quinoxaline and analogs.

The antiherpesvirus activity of 14 derivatives of indoloquinoxaline was tested. The most active was 2,3-dimethyl(dimethylaminoethyl)5H-indolo-(2,3-b)quinoxaline, also called B-220. The antiherpesvirus mechanism of B-220 was sought. The compound inhibited replication of herpes simplex virus type 1, cytomegalovirus, and varicella-zoster virus in tissue culture at concentrations of 1 to 5 microM, depending on the cell type used for assay and the amount of virus. Cellular toxicity was seen at a concentration of 10 to 30 microM, and antiviral activity in the human bladder cancer and human embryonic lung fibroblast cell lines tested was found at concentrations 3 to 15 times lower than the concentrations causing cellular toxicity. Viral DNA synthesis, as well as production of early and late viral proteins, was inhibited at 0.5 to 4.5 microM B-220, but viral DNA polymerases tested in vitro were not inhibited at these concentrations. There was no interaction with the pyrophosphate analog foscarnet, and no reversal of the antiviral activity of B-220 occurred with naturally occurring nucleosides. We conclude that the antiviral effect depends on the multiplicity of infection and may occur at the level of viral DNA synthesis and that no interference occurs with pyrophosphate analogs or nucleosides. The more potent activity against viral DNA than against cellular DNA may be caused by a true selectivity for herpesvirus DNA or by the higher metabolism of viral DNA in infected cells.