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2 protocols using 4 picoline

1

Characterization of P99 β-Lactamase Kinetics

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The ß-lactamase of Enterobacter cloacae P99 was obtained from the Centre for Applied Microbiology and Research (Porton Down, Wiltshire, U.K.) and used as supplied. The catalytic properties of this enzyme preparation have been previously documented and the concentrations of stock solutions were obtained spectrophotometrically [21 (link)]. Cephalothin, cephalosporin C, 7-aminocephalosporanic acid (ACA), and deacetoxycephalothin were gifts from Eli Lilly and Co. Cefoxitin was a gift from Merck & Co. (see Chart 1 for the substrate structures). Deuterium oxide (99.9 atom % 2H) was purchased from Sigma-Aldrich. The phosphonate inhibitors, p-nitrophenyl N-(methoxycarbonyl)aminomethylphosphonate (1), m-nitrophenyl N-(phenylacetyl)aminomethylphosphonate (2), p-nitrophenyl N-(benzyloxycarbonyl)aminomethylphosphonate (3), and benzoyl N-(benzyloxycarbonyl)aminomethylphosphonate (4) (See Chart 2) were available from previous studies in this laboratory [14 (link),22 (link)]. Imidazole and triethanolamine were purchased from Fluka Chemical Co. Thiazolidine, oxazole, 4-picoline and Tris were from Sigma-Aldrich and 2,2,2-trifluoroethylamine from Acros Organics.
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2

Synthesis and Biological Evaluation of Pyridine Derivatives

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2-Phenylpyridine, 4-pyridinemethanol, 4-dimethylaminopyridine,
methylnicotinate, N,N-diethylnicotinamide, 3-picoline,
4-picoline, 3-acetylpyridine, 9-ethylguanine, and 9-methyladenine
were purchased from Sigma-Aldrich. For the biological experiments,
RPMI-1640 medium, fetal bovine serum, l-glutamine, penicillin/streptomycin
mixture, trypsin/EDTA, and phosphate-buffered saline (PBS) were purchased
from PAA Laboratories GmbH. Cisplatin CDDP (≥99.9%), trichloroacetic
acid (≥99%), sulforhodamine B (75%), sodium phosphate monobasic
monohydrate (≥99%), sodium phosphate dibasic heptahydrate (≥99%),
acetic acid (≥99%), staurosporine, propidium iodide (>94%),
and RNase A were obtained from Sigma-Aldrich. Complex [(η5-Cpxph)Ir(phpy)Cl] was prepared according to reported
methods.6d
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