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33748253031
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note
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The absolute stereochemistry of the acetic acid side chain present in 35 and 36 was established by X-ray analysis of the following acid which was obtained from ester hydrolysis and resolution of 45: {A figure is presented} Crystallographic data (excluding structure factors) for the structures in this paper have been deposited with the Cambridge Crystallographic Data Centre as supplementary publication numbers CCDC 297169 (17) and CCDC 297363 (48). Copies of the data can be obtained, free of charge, on application to CCDC, 12 Union Road, Cambridge, CB2 1EZ, UK [e-mail: deposit@ccdc.cam.ac.uk].
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33748273075
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note
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125I]cAMP scintillation proximity assay (SPA) (RPA556, Amersham).
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13
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33748283926
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note
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TP functional assay was performed as follows: Test compounds were assessed for their ability to inhibit platelet aggregation in human PRP using aggregometry. Two hundred and fifty microliters of PRP was stirred in a cuvette at 37 °C for 3 min. Twenty-five microliters of the test compound diluted in buffer [0.9% (w/v) or 0.15 M NaCl containing 0.25% (w/v) BSA] or vehicle (buffer), as control was added to 250 μL stirring platelets for a 10 min preincubation period. Twenty microliters of agonist (1.36 μM U46619, final concentration) was then added to the stirring PRP mixture to induce aggregation and the incubation continued for five additional minutes. Maximum aggregation was determined at approximately 5 min. The maximum extent of aggregation achieved in the presence of U46619 was set to 100%.
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14
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33748250442
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note
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All new compounds provided satisfactory spectral data along with mass spectral and/or elemental analysis.
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0030908560
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