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Vol. 62, Issue 1, 58-64, July 2002
1-Induced
Extracellular Matrix with a Novel Inhibitor of the TGF- Type I
Receptor Kinase Activity: SB-431542
Departments of Renal and Urology Research (N.J.L., E.G., A.M.,
S.B., J.B., C.T., W.M., B.A.O.), Gene Expression Sciences (J.F.),
Protein Biochemistry (R.L.), and Medicinal Chemistry (J.H., L.G.,
J.F.C.), GlaxoSmithKline Pharmaceuticals, King of Prussia, Pennsylvania
Transforming growth factor 
1 (TGF-1) is a potent fibrotic factor
responsible for the synthesis of extracellular matrix. TGF-1 acts
through the TGF- type I and type II receptors to activate
intracellular mediators, such as Smad proteins, the p38 mitogen-activated protein kinase (MAPK), and the extracellular signal-regulated kinase pathway. We expressed the kinase domain of the TGF- type I receptor [activin receptor-like kinase (ALK)5] and the substrate, Smad3, and determined that SB-431542 is a selective inhibitor of Smad3 phosphorylation with an IC50 of 94 nM.
It inhibited TGF-1-induced nuclear Smad3 localization. The p38
mitogen-activated protein kinase inhibitors SB-203580 and SB-202190
also inhibit phosphorylation of Smad3 by ALK5 with IC50
values of 6 and 3 µM, respectively. This suggests that these p38 MAPK
inhibitors must be used at concentrations of less than 10 µM to
selectively address p38 MAPK mechanisms. However, the p38 MAPK
inhibitor SB-242235 did not inhibit ALK5. To evaluate the relative
contribution of Smad signaling and p38 MAPK signaling in
TGF-1-induced matrix production, the effect of SB-431542 was
compared with that of SB-242235 in renal epithelial carcinoma A498
cells. All compounds inhibited TGF-1-induced fibronectin (FN) mRNA,
indicating that FN synthesis is mediated in part via the p38 MAPK
pathway. In contrast, SB-431542, but not the selective p38 MAPK
inhibitor SB-242235, inhibited TGF-1-induced collagen I
1 (col
I1). These data indicate that some matrix markers that are
stimulated by TGF-1 are mediated via the p38 MAPK pathway (i.e.,
FN), whereas others seem to be activated via ALK5 signaling independent
of the p38 MAPK pathway (i.e., col I1).
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