Ii Masayuki, Matsunaga Naoko, Hazeki Kaoru, Nakamura Kazuyo, Takashima Katsunori, Seya Tsukasa, Hazeki Osamu, Kitazaki Tomoyuki, Iizawa Yuji
Pharmacology Research Laboratories I, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 17-85, Jusohonmachi 2-chome, Yodogawa-ku, Osaka 532-8686, Japan.
Mol Pharmacol. 2006 Apr;69(4):1288-95. doi: 10.1124/mol.105.019695. Epub 2005 Dec 22.
Proinflammatory mediators such as cytokines and NO play pivotal roles in various inflammatory diseases. To combat inflammatory diseases successfully, regulation of proinflammatory mediator production would be a critical process. In the present study, we investigated the in vitro effects of ethyl (6R)-6-[N-(2-chloro-4-fluorophenyl)sulfamoyl]cyclohex-1-ene-1-carboxylate (TAK-242), a novel small molecule cytokine production inhibitor, and its mechanism of action. In RAW264.7 cells and mouse peritoneal macrophages, TAK-242 suppressed lipopolysaccharide (LPS)-induced production of NO, tumor necrosis factor-alpha (TNF-alpha), and interleukin (IL)-6, with 50% inhibitory concentration (IC50) of 1.1 to 11 nM. TAK-242 also suppressed the production of these cytokines from LPS-stimulated human peripheral blood mononuclear cells (PBMCs) at IC50 values from 11 to 33 nM. In addition, the inhibitory effects on the LPS-induced IL-6 and IL-12 production were similar in human PBMCs, monocytes, and macrophages. TAK-242 inhibited mRNA expression of IL-6 and TNF-alpha induced by LPS and interferon-gamma in RAW264.7 cells. The phosphorylation of mitogen-activated protein kinases induced by LPS was also inhibited in a concentration-dependent manner. However, TAK-242 did not antagonize the binding of LPS to the cells. It is noteworthy that TAK-242 suppressed the cytokine production induced by Toll-like receptor (TLR) 4 ligands, but not by ligands for TLR2, -3, and -9. In addition, IL-1beta-induced IL-8 production from human PBMCs was not markedly affected by TAK-242. These data suggest that TAK-242 suppresses the production of multiple cytokines by selectively inhibiting TLR4 intracellular signaling. Finally, TAK-242 is a novel small molecule TLR4 signaling inhibitor and could be a promising therapeutic agent for inflammatory diseases, whose pathogenesis involves TLR4.
细胞因子和一氧化氮（NO）等促炎介质在多种炎症性疾病中起关键作用。为了成功对抗炎症性疾病，调节促炎介质的产生将是一个关键过程。在本研究中，我们研究了新型小分子细胞因子产生抑制剂（6R）-6-[N-（2-氯-4-氟苯基）氨磺酰基]环己-1-烯-1-羧酸乙酯（TAK-242）的体外作用及其作用机制。在RAW264.7细胞和小鼠腹腔巨噬细胞中，TAK-242抑制脂多糖（LPS）诱导的NO、肿瘤坏死因子-α（TNF-α）和白细胞介素（IL）-6的产生，50%抑制浓度（IC50）为1.1至11 nM。TAK-242还抑制LPS刺激的人外周血单核细胞（PBMC）产生这些细胞因子，IC50值为11至33 nM。此外，对LPS诱导的人PBMC、单核细胞和巨噬细胞中IL-6和IL-12产生的抑制作用相似。TAK-242抑制RAW264.7细胞中LPS和干扰素-γ诱导的IL-6和TNF-α的mRNA表达。LPS诱导的丝裂原活化蛋白激酶的磷酸化也以浓度依赖的方式受到抑制。然而，TAK-242并不拮抗LPS与细胞的结合。值得注意的是，TAK-242抑制Toll样受体（TLR）4配体诱导的细胞因子产生，但不抑制TLR2、-3和-9配体诱导的细胞因子产生。此外，TAK-242对IL-1β诱导的人PBMC中IL-8产生没有明显影响。这些数据表明，TAK-242通过选择性抑制TLR4细胞内信号传导来抑制多种细胞因子的产生。最后，TAK-242是一种新型小分子TLR4信号抑制剂，可能是治疗发病机制涉及TLR4的炎症性疾病的有前景的治疗剂。
