In experiments on cellular models, GI254023X markedly inhibits the baseline shedding of RAGE at a concentration of 25 μM, and this inhibitory effect persists, albeit to a lesser extent, at 1 μM. The compound shows a modest reduction in RAGE shedding at a concentration of 100 nM. Distinguishing between various proteinases in vitro, GI254023X exhibits selectivity, demonstrating an IC50 of 541 nM against ADAM17 and more potent inhibition against ADAM10 (IC50=5.3 nM) and MMP9 (IC50=2.5 nM)^[1].

The shedding of CXCL16 is obstructed by inhibitors of ADAM proteases, such as GI254023x. A2780 cells, when treated with the ADAM-10/ADAM-17 inhibitor TAPI-2 along with GI254023x, a selective inhibitor for ADAM-10, reveal a significant difference in the expression levels of ADAM-10 mRNA, being almost 9.8 times higher than that of ADAM-17. Moreover, GI254023x efficiently prevents the shedding of CXCL16 from the cell surface, surpassing the effectiveness of TAPI-2^[2].

Utilizing the specific inhibitor for ADAM10 (α-secretase), GI254023X, at a concentration of 5 mM on serum/glucose-depleted slices results in a noticeable rise in PI counts compared to slices treated with DMSO (the carrier)^[3].