Herein, two series of tryptanthrin derivatives with benzenesulfonamide substituents were designed and synthesized to discover novel anti-inflammatory agents. The anti-inflammatory activities of all derivatives were screened by evaluating their inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells. Among them, compound 8j exhibited the best NO inhibitory activity (IC₅₀ = 1.25 ± 0.21 μM), with no obvious toxicity. Further evaluation showed that 8j could also significantly reduce the levels of pro-inflammatory cytokines interleukin-1β (IL-1β, IC₅₀ = 8.48 ± 0.23 μM) and tumor necrosis factor-α (TNF-α, IC₅₀ = 11.53 ± 0.35 μM) and downregulate the LPS-induced expression of iNOS and COX-2. Reverse docking of 8j suggested p38α as the molecular target, which is a well-known crucial player in the p38 MAPK signaling pathway that controls the transcription of pro-inflammatory mediators. Cellular thermal shift assay showed that 8j efficiently stabilized p38α in LPS-treated RAW264.7 cells. Western blot showed that inflammatory response was inhibited by 8j through inhibiting the phosphorylation of p38α and MK2 in the p38 MAPK signaling pathway. Finally, In vivo studies showed that 8j could significantly ameliorate the degree of foot swelling and knee joint pathology in adjuvant-induced arthritis (AIA) rats and reduce levels of TNF-α and IL-1β in serum, achieving the effect of protecting synovial tissue and ameliorating arthritis. These findings suggested that 8j may be a promising compound for further development of anti-inflammatory agents.

在此，设计并合成了两个系列的带有苯磺酰胺取代基的色胺酮衍生物，以发现新型抗炎剂。通过评估所有衍生物对脂多糖（LPS）诱导的RAW264.7细胞中一氧化氮（NO）产生的抑制作用来筛选所有衍生物的抗炎活性。其中，化合物8j表现出最佳的NO抑制活性（IC ₅₀ = 1.25 ± 0.21 μM），且无明显毒性。进一步评估表明， 8j还可以显着降低促炎细胞因子白细胞介素-1β（IL-1β，IC ₅₀ = 8.48 ± 0.23 μM）和肿瘤坏死因子-α（TNF-α，IC ₅₀ = 11.53 ± 0.35 μM）的水平) 并下调 LPS 诱导的 iNOS 和 COX-2 表达。 8j的反向对接表明 p38α 作为分子靶标，它是控制促炎介质转录的 p38 MAPK 信号通路中众所周知的关键角色。细胞热位移测定表明， 8j有效稳定 LPS 处理的 RAW264.7 细胞中的 p38α。 Western blot显示8j通过抑制p38 MAPK信号通路中p38α和MK2的磷酸化来抑制炎症反应。最后，体内研究表明， 8j可显着改善佐剂性关节炎（AIA）大鼠的足部肿胀程度和膝关节病理，并降低血清中TNF-α和IL-1β的水平，达到保护滑膜组织的作用并改善关节炎。 这些发现表明8j可能是一种有前途的化合物，可用于进一步开发抗炎药物。