The relationship between TLR4 and inflammation-related diseases has been paid more and more attention. The studies have shown that TLR4/NF-κB signaling pathway plays an important role in the transmission of inflammatory signals. A large number of pro-inflammatory factors, chemokines, adhesion factors, TLR4 and its ligands interact with each other, and jointly promote the development of diseases. In this work, 8 target compounds were synthesized to screen the inhibitory activity of TLR4 in vitro. The results of TLR4 inhibition test in vitro showed that the double-ring conjugated enones had a good inhibitory activity, and the IC₅₀ value of compound 4f was 0.56 ± 0.10 μM, and it was superior to the positive control methotrexate. To further study the anti-inflammatory effect and mechanism of double-ring conjugated enones by using LPS induced rat synovial cell inflammation model. The results of the mechanism test showed that compound 4f could effectively promote the apoptosis of rat synovial cells, and the mechanism might be related to the up-regulation of the expression of apoptosis-related protein Caspase-3. In addition, compound 4f could significantly inhibit the increase of inflammatory factors TNF-α, IL-1β and IL-6 in rat synovial cells induced by LPS, showing a good anti-inflammatory activity. In the TLR4/NF-κB signaling pathway test of rat synovial cells, compound 4f can effectively regulate the expression levels of TLR4, MyD88, NF-κB and IκB related proteins in TLR4/NF-κB signaling pathway, which may be due to its inhibition of LPS-induced inflammation in rat synovial cells. At the same time, it inhibits the abnormal proliferation of cells and its important mechanism promoted of apoptosis.

TLR4与炎症相关疾病的关系越来越受到重视。研究表明，TLR4/NF-κB信号通路在炎症信号的传递中起重要作用。大量促炎因子、趋化因子、粘附因子、TLR4及其配体相互作用，共同促进疾病的发展。在这项工作中，合成了 8 个目标化合物，以在体外筛选 TLR4 的抑制活性。体外TLR4抑制试验结果表明，双环偶联烯酮具有良好的抑制活性，化合物4f的IC ₅₀值为 0.56 ± 0.10 μM，优于阳性对照甲氨蝶呤。采用LPS诱导的大鼠滑膜细胞炎症模型进一步研究双环偶联烯酮的抗炎作用及其机制。机制试验结果表明，化合物4f可有效促进大鼠滑膜细胞凋亡，其机制可能与上调凋亡相关蛋白Caspase-3的表达有关。此外，化合物4f可显着抑制LPS诱导的大鼠滑膜细胞炎症因子TNF-α、IL-1β和IL-6的升高，显示出良好的抗炎活性。在大鼠滑膜细胞的 TLR4/NF-κB 信号通路试验中，化合物4f能有效调节TLR4/NF-κB信号通路中TLR4、MyD88、NF-κB和IκB相关蛋白的表达水平，这可能与其抑制LPS诱导的大鼠滑膜细胞炎症有关。同时具有抑制细胞异常增殖和促进细胞凋亡的重要机制。