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
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
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DOI：
10.1124/mol.110.068064
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发表时间：
2011-01-01
影响因子：
3.6
通讯作者：
Ii, Masayuki
中科院分区：
文献类型：
--
作者：
Matsunaga, Naoko;Tsuchimori, Noboru;Ii, Masayuki
TAK-242 （resatorvid）是toll样受体（TLR） 4信号传导的小分子特异性抑制剂，通过与TLR4的胞内结构域结合，抑制脂多糖诱导的炎症介质的产生。TLR4中的Cys747先前已被确定为TAK-242的结合位点。然而，TAK-242与TLR4结合后抑制TLR4信号传导的机制尚不清楚。本研究利用共免疫沉淀证明，TAK-242干扰TLR4及其接头分子之间的蛋白相互作用。在10种不同的人类tlr中，TAK-242选择性地与TLR4结合。TAK-242对炎症介质产生抑制作用的时间过程与TAK-242与TLR4结合的时间过程一致。在过表达TLR4、MD-2、TIRAP或TRAM的人胚胎肾（HEK） 293细胞中，TAK-242分别抑制TLR4与Toll/白介素-1受体域衔接蛋白（TIRAP）或Toll/白介素-1受体域衔接蛋白诱导干扰素- β相关衔接分子（TRAM）的关联。在稳定表达TLR4、MD-2和CD14的HEK293细胞中，TAK-242抑制tirap介导的核因子κ B （nf - κ B）的激活和tram介导的nf - κ B和干扰素敏感反应元件的激活。TAK-242也抑制了RAW264.7细胞内源性白细胞介素-1受体相关激酶的激活。这些发现表明，TAK-242选择性地与TLR4结合，随后破坏TLR4与接头分子的相互作用，从而抑制TLR4信号转导及其下游信号事件。这项工作提出了一种能够破坏蛋白质-蛋白质相互作用的小分子的新范式。
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. Cys747 in TLR4 has been identified previously 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 10 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 Toll/interleukin-1 receptor domain-containing adaptor protein (TIRAP) or Toll/interleukin-1 receptor domain-containing adaptor protein inducing interferon-beta-related adaptor molecule (TRAM) in human embryonic kidney (HEK) 293 cells overexpressing TLR4, MD-2, and TIRAP or TRAM, respectively. TAK-242 inhibited the TIRAP-mediated activation of nuclear factor kappa B (NF-kappa B) and the TRAM-mediated activation of NF-kappa B 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.