4636 Journal of Medicinal Chemistry, 2006, Vol. 49, No. 15
Venkatesan et al.
N. J. P.; Calvert, S. H.; Coleman, K.; Francois, I. 6-(Substituted
Methylene) Penems: Potent Broad Spectrum Inhibitors of Bacterial
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(1.04 mL) in one portion. The reaction mixture was warmed to 0
°C and stirred for 15 h. The mixture was diluted with ethyl acetate
and washed with 5% citric acid aqueous solution, saturated sodium
hydrogen carbonate, and brine. The organic layer was dried
(MgSO4) and filtered through a pad of Celite. The pad was washed
with ethyl acetate. The filtrate was concentrated under reduced
pressure. The residue was applied to silica gel column chromatog-
raphy, and then the column was eluted with ethyl acetate/hexane
(1:1). The collected fractions were concentrated under reduced
pressure, and the mixture of diastereoisomers were taken to the
next step; pale yellow amorphous solid. Yield: 0.76 g (65%); (M
+ H)+ 579.
Step-5: (5R)(6Z)-6-(6,7-5H- Dihydropyrazolo[5,1-b]oxazin-2-
ylmethylene)-7-oxo-4-thia-1- azabicyclo[3.2.0]hept-2-ene-2-car-
boxylic Acid, Sodium Salt (14k). 4-Nitrobenzy(5R)-6-[(acetyloxy)-
(6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazin-2-yl)methyl]-6-bromo-
7-oxo-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (13k) (350
mg, 0.6 mmol) was dissolved in THF (20 mL), acetonitrile (10
mL), and 0.5 M phosphate buffer (pH 6.5, 28 mL) and hydrogenated
over 10% Pd/C at 40 psi pressure. After 4 h, the reaction mixture
was filtered and cooled to 3 °C, and 0.1 N NaOH was added to
adjust the pH to 8.5. The filtrate was washed with ethyl acetate,
and the aqueous layer was separated. The aqueous layer was
concentrated under high vacuum at 35 °C to give a yellow
precipitate. The product was purified by HP21 resin reverse-phase
column chromatography. Initially, the column was eluted with
deionized water (2 L) and the latter with 10% acetonitrile/water.
The fractions containing the product were collected and concentrated
at reduced pressure at room temperature. The yellow amorphous
solid was washed with acetone and filtered. Yield: 103 mg (52%).
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Acknowledgment. We thank the members of the Wyeth
Chemical Technologies group for the analytical and spectral
determination and Drs. Magid Abou-Gharbia, Patricia Bradford,
Beth Rasmussen, Steve Projan, and John O’Connell for useful
discussions. We also thank Drs. Howard Sard, Vishnu Hegde,
and Mario Gonzalez of Organix Inc., Wobern, MA for scaling
up compound 11.
Supporting Information Available: Elemental analysis data
and 1H NMR data for compounds 14a-k. This material is available
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