Identification of a novel 2-pyridyl-benzensulfonamide derivative, RQ-00203078, as a selective and orally active TRPM8 antagonist.

A novel series of 2-pyridyl-benzensulfonamide derivatives have been identified as selective and orally active TRPM8 antagonists via high throughput screening (HTS). Exploration of the structure-activity relationships of compound 1 has led to the identification of RQ-00203078 (compound 36) as a highly selective, potent and orally available TRPM8 antagonist. RQ-00203078 demonstrated excellent in vivo activity in a dose dependent manner with an ED50 value of 0.65 mg/kg in the icilin-induced wet-dog shakes model in rats after oral administration and may become an important pharmacological tool for fully assessing the potential therapeutic use of the targets activated by cold stimulation.

Foaming control by automatic carbon source adjustment using an ORP profile in sequencing batch reactors for enhanced nitrogen removal in swine wastewater treatment.

Incomplete denitrification and ammonia accumulation were found to cause proliferation of filamentous microorganisms in sequencing batch reactors (SBRs) for swine wastewater treatment. Foaming was observed in response to the accumulation of 115.2 and 12.2 mg/L of nitrate and ammonia, respectively. The mixed liquor suspended solids (MLSS) level in SBRs was decreased to 2,000 mg/L and the suspended solids in the effluent reached 200 mg/L when foaming appeared. However, the use of swine waste as an external carbon source for enhanced biological nitrogen removal was found to effectively control the foaming caused by filamentous microorganisms. Therefore, an optimum strategy for the addition of swine waste was designed using integrated real-time control to provide pulse input control of slurry based on the "nitrate knee" in the oxidation-reduction potential profile. In this case, the MLSS concentration was maintained at an average value of approximately 7,550 mg/L, while the SS in the effluent was less than 30 mg/L.

Reactive oxygen species assay-based risk assessment of drug-induced phototoxicity: classification criteria and application to drug candidates.

We have previously demonstrated that the phototoxic potential of chemicals could be partly predicted by the determination of reactive oxygen species (ROS) from photo-irradiated compounds. In this study, ROS assay strategy was applied to 39 marketed drugs and 210 drug candidates in order to establish provisional classification criteria for risk assessment of drug-induced phototoxicity. The photosensitizing properties of 39 model compounds consisting of phototoxic and non-phototoxic chemicals, as well as ca. 210 drug candidates including 11 chemical series were evaluated using ROS assay and the 3T3 neutral red uptake phototoxicity test (NRU PT). With respect to marketed drugs, most phototoxic drugs tended to cause type I and/or II photochemical reactions, resulting in generation of singlet oxygen and superoxide. There seemed to be a clear difference between phototoxic drugs and non-phototoxic compounds in their abilities to induce photochemical reactions. A plot analysis of ROS data on the marked drugs provided classification criteria to discriminate the photosensitizers from non-phototoxic substances. Of all drug candidates tested, 35.2% compounds were identified as phototoxic or likely phototoxic on the basis of the 3T3 NRU PT, and all ROS data for these phototoxic compounds were found to be over the threshold value. Furthermore, 46.3% of non-phototoxic drug candidates were found to be in the subthreshold region. These results verify the usefulness of the ROS assay for understanding the phototoxicity risk of pharmaceutical substances, and the ROS assay can be used for screening purposes in the drug discovery stage.

Mass balance study of nonylphenol ethoxylates and their metabolites in an urban river contaminated by nonylphenol.

In previous studies, an urban river called Kamo River was found to be polluted with nonylphenol (NP) by a rubber product manufacturing factory. To determine the contribution of the factory effluent to the river pollution, a mass balance study was conducted for NP, nonylphenol ethoxylates (NPnEOs, n = 1-15), and nonylphenol carboxylates (NPmECs, m = 1-10) in the waters of the river and its 10 inflow channels. The total concentration of nonylphenolic compounds (T-NPCs) in the river water was 4.6 nM/L at the upstream sampling point and 54.6 nM/L at the downstream sampling point. T-NPCs concentration ranged from 0.4 to 487.1 nM/L in the waters of the inflow channels connected to the targeted river section, and the highest value was observed in the water of the inflow channel which receives the factory effluent. The mass flows of T-NPCs were 458.0 mM/day for Input (the upstream sampling point and outlets of 10 inflow channels) and 828.2 mM/day for Output (the downstream sampling point). The mass flow of the highly polluted inflow channel comprised 82.4% of Input. In comparison with the mass balance of chloride ion, the mass flow of T-NPCs in Output was still higher than that in Input. This phenomenon was attributed to the high mass flows of NP and NPnEOs (n = 1-3) in Output, and their potential source was determined to be the river sediment.

Seasonal changes in nonylphenol ethoxylates and their metabolites in water and sediment of urban river polluted by nonylphenol.

The concentrations of nonylphenol (NP), nonylphenol ethoxylates (NPnEOs: n = 1-15) and nonylphenoxy acetic acids (NPmECs: m = 1-10) in river water and sediment were determined seasonally at four sampling sites in the Kamo River, which is a typical urban river and was already known to be polluted by NP. In the water, the concentrations of NP, NPnEOs and NPmECs ranged from 0.13 to 3.65 microg/L, from 1.0 to 22.4 microg/L, and from 0.6 to 8.5 microg/L, respectively. NP, nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), nonylphenoxy acetic acid (NP1EC) and nonylphenol monoethoxy acetic acid (NP2EC) were predominant in the water. In the sediment, the concentrations of NP, NPnEOs and NPmECs ranged from 69 to 10,747 microg/kg dry weight (dw), from 74 to 6,101 microg/kg dw, and from 24 to 673 microg/kg dw, respectively. NP, NP1EO and NP2EO were predominant in the sediment. The total concentrations of NP, NPnEOs and NPmECs in both river water and sediment were higher in winter and spring than in summer and autumn. In the river water, the ratio in concentration of the sum of NP, NP1EO, NP2EO, NP1EC and NP2EC to the whole nonylphenolic compounds was correlated with water temperature. The surveyed results suggested that some point emission sources of nonylphenolic compounds were located in the river basin between the first sampling site and the second sampling site.

Effects of 5-HT2B, 5-HT3 and 5-HT4 receptor antagonists on gastrointestinal motor activity in dogs.

AIM: To study the effects of 5-hydroxytryptamine (5-HT) receptor antagonists on normal colonic motor activity in conscious dogs. METHODS: Colonic motor activity was recorded using a strain gauge force transducer in 5 dogs before and after 5-HT2B, 5-HT3 and 5-HT4 receptor antagonist administration. The force transducers were implanted on the serosal surfaces of the gastric antrum, terminal ileum, ileocecal sphincter and colon. Test materials or vehicle alone was administered as an intravenous bolus injection during a quiescent period of the whole colon in the interdigestive state. The effects of these receptor antagonists on normal gastrointestinal motor activity were analyzed. RESULTS: 5-HT2B, 5-HT3 and 5-HT4 receptor antagonists had no contractile effect on the fasting canine terminal ileum. The 5-HT3 and 5-HT4 receptor antagonists inhibited phase III of the interdigestive motor complex of the antrum and significantly inhibited colonic motor activity. In the proximal colon, the inhibitory effect was dose dependent. Dose dependency, however, was not observed in the distal colon. The 5-HT2B receptor antagonist had no contractile effect on normal colonic motor activity. CONCLUSION: The 5-HT3 and 5-HT4 receptor antagonists inhibited normal colonic motor activity. The 5-HT2B receptor antagonist had no contractile effect on normal colonic motor activity.

A simplified determination method of dioxin toxic equivalent (TEQ) by single GC/MS measurement of five indicative congeners.

Here, we proposed a simplified determination method for dioxin (polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls (PCBs)) toxic equivalency (TEQ; based on the World Health Organization-2006 toxicity equivalent factor) in environmental samples collected in Japan. This method has estimated the total TEQ from the concentrations of only five indicative congeners, namely, 2,3,4,7,8-pentachlorodibenzofuran; 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin; 1,2,3,7,8-pentachlorodibenzo-p-dioxin; and PCBs of IUPAC Nos. #126 and #105, which were obtained from a single GC/MS measurement using a combination of columns of 10-meter-long DB-5ms (injector side) and 20-meter-long DB-17ms (detector side). Applying this simplified determination method to various environmental samples (ambient air, river water, river sediment, and soil) showed that the estimated TEQ was in good agreement with TEQ obtained by the corresponding official methods. Moreover, this method could estimate the TEQ contributions of the four major dioxin sources in Japan: combustion by-products, pentachlorophenol formulations, chlornitrofen formulations and PCB products.

Seasonal change of PCDDs/PCDFs/DL-PCBs in the water of Ayase River, Japan: pollution sources and their contributions to TEQ.

In Japan, Ayase River is one of the most polluted rivers by PCDDs, PCDFs and dioxin-like PCBs, which are referred to as dioxins in this paper. The water samples of the river were collected once per month for a year, and dioxins were analyzed to examine the dioxin sources and their contributions to toxic equivalent (TEQ). The WHO-2006 TEQs ranged from 0.26 to 7.0 pg-TEQL(-1) and the average was 2.7 pg-TEQL(-1); eight of 12 samples exceeded the environmental quality standard in Japan (1.0 pg-TEQL(-1)). The TEQ value was high during the irrigation period from May to August. The most part of the dioxins in the river water existed in suspended solids (SS) and it seemed that the river received water with highly-dioxin-contaminated SS in the irrigation period. The homologue profiles of the water samples suggested that the dioxins were influenced by pentachlorophenol (PCP) and chlornitrofen (CNP) formulations which were widely used as herbicides for the paddy fields in Japan. According to TEQ apportionment estimated by using indicative congeners, the TEQ was mainly contributed by PCP. Moreover, it was also shown that the TEQ contributions of PCP and CNP formulations increased along with the increase of the total TEQ and the TEQ contribution was dominated by these herbicides during the irrigation period. Therefore, it was concluded that the herbicides-originated dioxins run off from the paddy fields into the river during the irrigation period and increased the dioxins level in the river water. The result from the principal component analysis was consistent with these conclusions.

Polychlorinated dibenzo-p-dioxins, dibenzofurans, and dioxin-like polychlorinated biphenyls in rice straw smoke and their origins in Japan.

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (DL-PCBs) contained in the smoke generated from rice straw burning in post-harvest paddy fields in Japan were analyzed to determine their congener profiles. Both the apportionment of toxic equivalent (TEQ) by using indicative congeners and the comparison of the homolog profiles showed that the PCDDs/PCDFs/DL-PCBs present in the rice-straw smoke were greatly influenced by those present as impurities in pentachlorophenol (PCP) and chlornitrofen (CNP, 4-nitrophenyl-2,4,6-trichlorophenyl ether) formulations that had been widely used as herbicides in paddy fields in Japan. Further, in order to investigate the effects of paddy-field soil on the PCDDs/PCDFs/DL-PCBs present in rice-straw smoke, PCDD/PCDF/DL-PCB homolog profiles of rice straw, rice-straw smoke and paddy-field soil were compared. Rice-straw smoke was generated by burning rice straw on a stainless-steel tray in a laboratory. The results suggested that the herbicides-originated PCDDs/PCDFs/DL-PCBs and the atmospheric PCDDs/PCDFs/DL-PCBs contributed predominantly to the presence of PCDDs/PCDFs/DL-PCBs in the rice-straw smoke while the contribution of PCDDs/PCDFs/DL-PCBs formed during rice straw burning was relatively minimal. The major sources of the PCDDs/PCDFs/DL-PCBs found in the rice-straw smoke were attributed primarily to the paddy-field soil adhered to the rice straw surface and secondarily to the air taken by the rice straw. The principal component analysis supported these conclusions. It is concluded that rice straw burning at paddy fields acts as a driving force in the transfer of PCDDs/PCDFs/DL-PCBs from paddy-field soil to the atmosphere.

Apportionment of TEQs from four major dioxin sources in Japan on the basis of five indicative congeners.

The major sources of dioxins (polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (DL-PCBs)) in the environment in Japan have been considered to be combustion by-products, pentachlorophenol (PCP) formulations, chlornitrofen (CNP, 4-nitrophenyl-2,4,6-trichlorophenyl ether) formulations, and PCB products. Data on PCDDs, PCDFs and DL-PCBs from the four sources were analyzed, and indicative congeners whose concentrations were highly correlated with WHO-2006 toxic equivalencies (TEQs) were identified for each source sample. The indicative congeners for combustion by-products, PCP formulations, and CNP formulations were 2,3,4,7,8-pentachlorodibenzofuran, 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin, and 1,2,3,7,8-pentachlorodibenzo-p-dioxin, respectively; for PCB products, the indicative congeners were IUPAC Nos. #126- and #105-pentachlorobiphenyls. Moreover, using the data on PCDDs, PCDFs and DL-PCBs, we developed a set of equations for estimating the apportionment of TEQs from the four sources by using only the concentrations of the above-mentioned five indicative congeners. The equations were used along with the analysis results of different types of environmental samples collected from Japan, to determine the TEQ contributions of the four sources. The obtained values of TEQ contributions seemed to be reasonable. The estimation method was developed by using the data on major dioxin sources in Japan, and therefore, it is generally adaptable to environmental samples from any part of Japan. The method may be usable for regions outside Japan if source identification is carried out and the estimation equations are modified appropriately.

Optimization of imidazole 5-lipoxygenase inhibitors and selection and synthesis of a development candidate.

Structural modification of imidazole 5-lipoxygenase (5-LO) inhibitors for optimizing inhibitory potency, pharmacokinetic behavior and toxicity (ocular) profile led to 4-{3-[4-(2-methyl-1H-imidazol-1-yl)phenylthio]}phenyl-3,4,5,6-tetrahydro-2H-pyran-4-carboxamide (6) with no observable ocular toxicity. The orally active and safe imidazole 5-LO inhibitor 6 was selected as a clinical candidate and advanced to clinical studies. An improved synthesis of 6 is also discussed.

Preparation of optically pure propargylic and allylic alcohols from 2-(trimethylsilyl)vinyl sulfoxides as a chiral ethynyl anion synthon: computational studies on elimination reaction of 2-(trimethylsilyl)vinyl sulfoxides.

The reaction of the alpha-carbanion derived from (trimethylsilyl)vinyl sulfoxides with aldehydes afforded a diastereomeric mixture of the products. Each diastereomer was subjected to specific elimination reactions to give optically pure propargylic, trimethylsilylated propargylic, and allylic alcohols. Acceleration of the sulfenic acid-elimination from the beta-silylvinyl sulfoxide was demonstrated by the ab initio calculation to be ascribed mainly to the beta-effect of the silyl group.