PT  - JOURNAL ARTICLE
AU  - Matsunaga, Naoko
AU  - Tsuchimori, Noboru
AU  - Matsumoto, Tatsumi
AU  - Ii, Masayuki
TI  - TAK-242 (Resatorvid), a Small Molecule Inhibitor of Toll-Like Receptor (TLR) 4 Signaling, Binds Selectively to TLR4 and Interferes with Interactions between TLR4 and Its Adaptor Molecules
AID  - 10.1124/mol.110.068064
DP  - 2010 Sep 29
TA  - Molecular Pharmacology
PG  - mol.110.068064
4099  - http://molpharm.aspetjournals.org/content/early/2010/09/29/mol.110.068064.short
4100  - http://molpharm.aspetjournals.org/content/early/2010/09/29/mol.110.068064.full
AB  - TAK-242 (resatorvid), a small molecule specific inhibitor of Toll-like receptor (TLR) 4 signaling, inhibits the production of lipopolysaccharide-induced inflammatory mediators by binding to the intracellular domain of TLR4. Previously, Cys747 in TLR4 was identified as the binding site of TAK-242. However, the mechanism by which TAK-242 inhibits TLR4 signaling after binding to TLR4 remains unknown. The present study demonstrated, using coimmunoprecipitation, that TAK-242 interferes with protein-protein interactions between TLR4 and its adaptor molecules. Among ten different human TLRs, TAK-242 selectively bound to TLR4. The time course of the inhibitory effect of TAK-242 on inflammatory mediator production corresponded to that of the binding of TAK-242 to TLR4. TAK-242 inhibited the association of TLR4 with TIR domain-containing adaptor protein (TIRAP) or TRIF-related adaptor molecule (TRAM) in HEK293 cells overexpressing TLR4, MD-2 and TIRAP or TRAM, respectively. TAK-242 inhibited the TIRAP-mediated activation of nuclear factor kappa B (NF-kB) and the TRAM-mediated activation of NF-kB and interferon-sensitive response element in HEK293 cells stably expressing TLR4, MD-2 and CD14. The activation of endogenous interleukin-1 receptor-associated kinase in RAW264.7 cells was also inhibited by TAK-242 treatment. These findings suggest that TAK-242 binds selectively to TLR4 and subsequently disrupts the interaction of TLR4 with adaptor molecules, thereby inhibiting TLR4 signal transduction and its downstream signaling events. This work proposes a novel paradigm of a small molecule capable of disrupting protein-protein interactions.