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.

中文翻译：

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新型环己烯衍生物乙基（6R）-6- [N-（2-氯-4-氟苯基）氨磺酰基]环己-1-烯-1-羧酸盐（TAK-242）选择性抑制通行费受体4介导通过抑制细胞内信号传导产生细胞因子。

促炎介质，例如细胞因子和NO在各种炎性疾病中起关键作用。为了成功地抵抗炎性疾病，调节促炎性介质的产生将是关键的过程。在本研究中，我们调查了新型的（6R）-6- [N-（2-氯-4-氟苯基）氨磺酰基]环己-1-烯-1-羧酸（TAK-242）的体外作用小分子细胞因子产生抑制剂及其作用机理。在RAW264.7细胞和小鼠腹膜巨噬细胞中，TAK-242抑制了脂多糖（LPS）诱导的NO，肿瘤坏死因子α（TNF-alpha）和白介素（IL）-6的产生，抑制浓度为50％（IC50 ）1.1至11 nM。TAK-242还抑制了LPS刺激的人外周血单核细胞（PBMC）产生的这些细胞因子，IC50值为11至33 nM。此外，在人PBMC，单核细胞和巨噬细胞中，对LPS诱导的IL-6和IL-12产生的抑制作用相似。TAK-242抑制LPS和γ-干扰素诱导的RAW264.7细胞中IL-6和TNF-α的mRNA表达。LPS诱导的有丝分裂原活化蛋白激酶的磷酸化也受到浓度依赖性的抑制。但是，TAK-242不能拮抗LPS与细胞的结合。值得注意的是，TAK-242抑制了由Toll样受体（TLR）4配体诱导的细胞因子生成，但没有抑制TLR2，-3和-9的配体诱导的细胞因子生成。另外，TAK-242不会明显影响人PBMC的IL-1β诱导的IL-8产生。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是一种新型的小分子TLR4信号抑制剂，可能是炎症疾病的有前途的治疗剂，其发病机理涉及TLR4。