2-Phenyl-9H-purine-6-carbonitrile derivatives as selective cathepsin S inhibitors.

Starting from previously disclosed equally potent cathepsin K and S inhibitor 4-propyl-6-(3-trifluoromethylphenyl)pyrimidine-2-carbonitrile 1, a novel 2-phenyl-9H-purine-6-carbonitrile scaffold was identified to provide potent and selective cathepsin S inhibitors.

Optimisation of 2-cyano-pyrimidine inhibitors of cathepsin K: improving selectivity over hERG.

Several structure-guided optimisation strategies were explored in order to improve the hERG selectivity profile of cathepsin K inhibitor 1, whilst maintaining its otherwise excellent in vitro and in vivo profile. Ultimately, attenuation of clogP and pK(a) properties proved a successful approach and led to the discovery of a potent analogue 23, which, in addition to the desired selectivity over hERG (>1000-fold), displayed a highly attractive overall profile.

Design and optimization of a series of novel 2-cyano-pyrimidines as cathepsin K inhibitors.

Morphing structural features of HTS-derived chemotypes led to the discovery of novel 2-cyano-pyrimidine inhibitors of cathepsin K with good pharmacokinetic profiles, for example, compound 20 showed high catK potency (IC(50)=4nM), >580-fold selectivity over catL and catB, and oral bioavailability in the rat of 52%.

Effects of postharvest preparation on organophosphate insecticide residues in apples.

Apples were sampled directly from orchard trees at 96, 45, and 21 days postapplication with one of three organophosphate insecticides (azinphos methyl, phosalone, or phosmet, respectively). Individual apples were prepared for analysis following one of three postharvest preparations: no preparation, rinsed with deionized water for 10-15 s, or rinsed and peeled. Azinphos methyl, phosalone, and phosmet concentrations ranged from below the level of detection to 5.26 ng/g, 94.7 to 5720 ng/g, and 0.011 to 663 ng/g in the apples that received no postharvest preparation, respectively. Although rinsed apples had lower maximum concentrations than observed in apples with no preparation, levels were not significantly lower. Concentrations of all three OP insecticides in apples that were rinsed followed by peeling, however, were much lower (below detection limits to 0.733 ng/g, azinphos methyl; 0.322-219 ng/g, phosalone; and below detection limits to 44.0 ng/g, phosmet) than observed in apples that had been rinsed alone. Rinsing and peeling of apples resulted in a 74.5-97.9% reduction in OP residues, while rinsing alone lowered mean concentrations by 13.5-28.7% relative to apples that received no postharvest preparation.

Multiresidue determination of quinolone and fluoroquinolone antibiotics in fish and shrimp by liquid chromatography/tandem mass spectrometry.

A multiresidue method was developed to measure low levels of 8 fluoroquinolones (norfloxacin, ofloxacin, danofloxacin, ciprofloxacin, desethylene ciprofloxacin, enrofloxacin, sarafloxacin, and difloxacin) and 4 quinolones (oxolinic acid, flumequine, nalidixic acid, and piromidic acid). Method detection limits range from 0.1 ng/g for quinolones to 0.4 ng/g for fluoroquinolones. Average recoveries range from 57 to 96%, depending on analyte and commodity; relative standard deviations are all less than 18%. The drugs are extracted from tissues using a mixture of ethanol and 1% acetic acid, diluted in aqueous HCI, and defatted by extraction with hexane. The compounds are further isolated using cation-exchange solid-phase extraction and measured using liquid chromatography with electrospray tandem mass spectrometry detection. The method has been evaluated and applied to the analysis of salmon, trout, and shrimp. Detectable residues were observed in 10 out of 73 samples, at concentrations ranging from 0.28 to 16 ng/g.

Discovery and development of 5-[(5S,9R)-9-(4-cyanophenyl)-3-(3,5-dichlorophenyl)-1-methyl-2,4-dioxo-1,3,7-triazaspiro[4.4]non-7-yl-methyl]-3-thiophenecarboxylic acid (BMS-587101)--a small molecule antagonist of leukocyte function associated antigen-1.

LFA-1 (leukocyte function-associated antigen-1), is a member of the beta2-integrin family and is expressed on all leukocytes. This letter describes the discovery and preliminary SAR of spirocyclic hydantoin based LFA-1 antagonists that culminated in the identification of analog 8 as a clinical candidate. We also report the first example of the efficacy of a small molecule LFA-1 antagonist in combination with CTLA-4Ig in an animal model of transplant rejection.

Organophosphate levels in apple composites and individual apples from a treated Canadian orchard.

Azinphos-methyl, phosalone, and phosmet were applied individually to separate rows of trees within a commercial apple orchard in Quebec, Canada, during the 2003 agricultural season. Apples were collected for residue analysis immediately prior to the harvesting of the remaining apples for market distribution and were prepared for analysis as both individual apples and as composites of eight individuals. Analysis of the three applied compounds, as well as five organophosphate insecticides that were not applied, was performed using gas chromatography-mass spectrometry. Azinphos-methyl, phosalone, and phosmet, which were applied, were detected in all samples analyzed at concentrations ranging from 0.004 ng/g to 2260 ng/g. Methidathion was not observed in any sample. Chlorpyrifos, diazinon, dimethoate, and malathion concentrations ranged from below method detection limits to 0.71 ng/g, and the detection frequency for these compounds ranged from 20% to 100%. Residues measured in this study were all below the Canadian maximum residue limit for apples. Variability factors ranged from 2 to 19 for all compounds observed in this study. Composite samples may not accurately reflect the extremes of exposure from consumption of single servings of apples.

Lead, cadmium and aluminum in Canadian infant formulae, oral electrolytes and glucose solutions.

Lead (Pb), cadmium (Cd) and aluminum (Al) were determined in 437 individual samples of infant formulae, oral electrolytes and 5% glucose solutions available in Canada. In the electrolytes, Cd and Pb concentrations were all below 0.01 and 0.041 ng g(-1), respectively. In the 5% glucose solutions, Pb and Cd levels averaged 0.01 and 0.09 ng g(-1), respectively. Reported on an as-consumed basis, Pb levels in milk- and soya-based formulae averaged 0.90 and 1.45 ng g(-1), respectively, while Cd levels averaged 0.23 and 1.18 ng g(-1), respectively Average Al levels on an as-consumed basis were 440 ng g(-1) (range 10-3400 ng g(-1)) in milk-based formulae and 730 ng g(-1) (range 230-1100 ng g(-1)) in soy-based formulae. Al concentrations increased in the following order: plain formula < low-iron formula < iron-supplemented formula < casein hydrolysate formula ≈ premature formula ≤ soy formula. For example, in the powdered formulae, average Al concentrations were 18 ng g(-1) for plain milk-based, 37 ng g(-1) for low-iron, 128 ng g(-1) for iron supplemented, 462 ng g(-1) for lactose-free, 518 ng g(-1) for hypoallergenic and 619 ng g(-1) for soy-based formula. Al concentrations, as-consumed, increased with decreasing levels of concentration: powder < concentrated liquid < ready-to-use. Formulae stored in glass bottles contained between 100 and 300 ng g(-1) more Al than the same formulae stored in cans. The source of the increased Al did not appear to be the glass itself, because most electrolytes and glucose solutions, also stored in glass, contained less than 8 ng g(-1) Al. Corresponding differences in Pb and Cd levels were not observed. Al concentrations varied substantially among manufacturers; however, all manufacturers were able to produce plain milk-based formulae containing less than 50 ng g(-1) Al, i.e. within the range of Al concentrations found in human milk. Next to soya-based and hypoallergenic formulae, premature formulae contained among the highest concentrations of Al, ranging 851-909 ng g(-1) from one manufacturer and 365-461 ng g(-1) from another.

Variability in captan residues in apples from a Canadian orchard.

Apple trees in an orchard in Quebec, Canada were treated, following label directions, with the fungicide captan (1,2,3,6-tetrahydro-N-(trichloromethylthio)phthalimide) during the 2003 agricultural season. A total of 142 apples from three rows of trees were selected for determination of captan by GC/MS. Individual apples were found to contain captan levels ranging from 16.9 to 6350 ng g-1. Only two individual apple samples exceeded the Canadian maximum residue limit (5000 ng g-1) for captan in apples. Six composite samples, comprising half portions of eight individual apples, were analysed from each of the three experimental rows. Composite samples ranged in concentration from 166 to 2620 ng g-1. The greatest uncertainty associated with the measured concentrations was due to variability among apples rather than the measurement of residue levels.