Figure 1 - full size


Figure 1.
Fig. 1. (A) Chemical structure of the small molecule TNF- inhibitor. See (12) for synthetic route used. (B) Compound inhibition of TNF- binding to TNFR1 in vitro. An ELISA (12) was used to measure inhibition of solution-phase TNF-R1 horseradish peroxidase conjugate binding to biotinylated TNF- immobilized on a strepavidin-coated microtiter plate by serial dilutions of compound. The solid line represents a four-parameter curve fit (20) that yielded an IC[50] value of 22 µM. (C) Compound inhibition of TNF- induced I B- depletion in HeLa cells. Cells were treated with sufficient TNF- or IL-1ß to give an 80% of maximal I B- depletion response after a 30-min exposure (0.4 and 0.04 ng/ml, respectively), as measured in an assay of cell lysates (12). Solid circles show that compound addition inhibits this TNF- -induced I B- depletion and yields an IC[50] value of 4.6 µM. Open circles show that compound addition does not affect orthogonal IL-1ß-induced I B- depletion. Error bars indicate standard deviations for triplicate measurements.

The above figure is reprinted by permission from the AAAs: Science (2005, 310, 1022-1025) copyright 2005.