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69949101429
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For kinetic off rate measurements, Jurkat cells were first treated with saturating levels of compound at 37 °C. After one hour incubation, the cells were collected by filtration and washed once with ice-cold binding buffer. Subsequently, cells were re-suspended in the binding buffer containing an excess of [125I]-tracer, and further incubated for increasing periods of time. The cells were collected at different time points, and the radioactivity associated with the cells was measured in a Packard TopCount NXTTM.
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For kinetic off rate measurements, Jurkat cells were first treated with saturating levels of compound at 37 °C. After one hour incubation, the cells were collected by filtration and washed once with ice-cold binding buffer. Subsequently, cells were re-suspended in the binding buffer containing an excess of [125I]-tracer, and further incubated for increasing periods of time. The cells were collected at different time points, and the radioactivity associated with the cells was measured in a Packard TopCount NXTTM.
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15
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27644441529
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Sandborn W.J., Colombel J.F., Enns R., Feagan B.G., Hanauer S.B., Lawrance I.C., Panaccione R., Sanders M., Schreiber S., Targan S., Deventer S.V., Goldblum R., Despain D., Hogge G.S., and Rutgeerts R. NEJM 353 (2005) 1912
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69949087328
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See Ref. 6a
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See Ref. 6a.
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18
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0038137501
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Leone D.R., Giza K., Gill A., Dolinski B.M., Yang W., Perper S., Scott D.M., Lee W.-C., Cornebise M., Wortham K., Nickerson-Nutter C., Chen L.L., LePage D., Spell J.C., Whalley E.T., Petter R.C., Adams S.P., Lobb R.R., and Pepinsky R.B. J. Pharmacol. Exp. Ther. 305 (2003) 1150
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19
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69949098975
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note
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HPLC log D also did not account for differences in binding affinity (all had similar values of 2 at pH 7.3).
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20
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69949108425
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note
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All the final compounds were greater than 95% ee as measured by chiral HPLC using analytical column (CHIRALCEL OD 10 μ, 4.6 × 250 mm) with heptane and isoproponol/triethylamine as mobile system.
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