Fang Shuopo, Huang Xiaodan, Cai Fen, Qiu Guodong, Lin Fei, Cai Xiaorui
Department of Pharmacy, The Affiliated Cancer Hospital of Shantou University Medical College, Shantou 515041, Guangdong, China.
Department of Digestive Medical Oncology, The Affiliated Cancer Hospital of Shantou University Medical College, Shantou 515041, Guangdong, China.
J Steroid Biochem Mol Biol. 2024 Jun;240:106478. doi: 10.1016/j.jsbmb.2024.106478. Epub 2024 Feb 29.
Inflammation, an important biological protective response to tissue damage or microbial invasion, is considered to be an alarming signal for the progress of varied biological complications. Based on the previous reports in the literature that proved the noticeable efficacy of pyrazole and thiazole scaffold as well as nitrogen heterocyclic based compounds against acute and chronic inflammatory disease, a new set of novel D-ring substituted steroidal 4,5-dihydropyrazole thiazole derivatives were synthesized and evaluated their anti-inflammatory activities in vitro. Preliminary structure-activity relationship (SAR) analysis was conducted by their inhibitory activities against nitric oxide (NO) release in lipopolysaccharide (LPS)-induced RAW 264.7 cells, and the optimal compound 12b [3β-hydroxy-pregn-5-en-17β-yl-5'- (o- chlorophenyl)- 1'-(4''- phenyl -[1'', 3'']- thiazol-2''- yl) - 4',5'-dihydro - 1'H-pyrazol - 3'- yl] exhibited more potent anti-inflammatory activity than the positive control treatment methylprednisolone (MPS), with an IC value of 2.59 μM on NO production and low cytotoxicity against RAW 264.7 cells. In further mechanism study, our results showed that compound 12b significantly suppressed the production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inhibited the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) through blocking NF-κB p65 nuclear translocation and phosphorylation of IκBα. Compound 12b also attenuated LPS-induced activation of c-Jun amino-terminal kinase (JNK) and p38 phosphorylation in RAW 264.7 cells. Molecular docking study revealed the strong binding affinity of compound 12b to the active site of the COX-2 proteins, which confirmed that compound 12b acted as an anti-inflammatory mediator. These results indicate that steroidal derivatives bearing 4,5-dihydropyrazole thiazole structure might be considered for further research and scaffold optimization in designing anti-inflammatory drugs and compound 12b might be a promising therapeutic anti-inflammatory drug candidate.
炎症是机体对组织损伤或微生物入侵的一种重要生物保护反应，被认为是各种生物并发症进展的警示信号。基于文献中先前的报道，证明了吡唑和噻唑骨架以及含氮杂环化合物对急慢性炎症性疾病具有显著疗效，我们合成了一组新型的D环取代甾体4,5-二氢吡唑噻唑衍生物，并在体外评估了它们的抗炎活性。通过它们对脂多糖（LPS）诱导的RAW 264.7细胞中一氧化氮（NO）释放的抑制活性进行了初步的构效关系（SAR）分析，最佳化合物12b [3β-羟基-孕甾-5-烯-17β-基-5'-（邻氯苯基）-1'-（4''-苯基-[1'', 3'']-噻唑-2''-基）-4',5'-二氢-1'H-吡唑-3'-基]表现出比阳性对照药物甲基泼尼松龙（MPS）更强的抗炎活性，对NO产生的IC值为2.59 μM，对RAW 264.7细胞的细胞毒性较低。在进一步的机制研究中，我们的结果表明化合物12b显著抑制肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）的产生，并通过阻断NF-κB p65核转位和IκBα磷酸化抑制诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）的表达。化合物12b还减弱了LPS诱导的RAW 264.7细胞中c-Jun氨基末端激酶（JNK）的激活和p38磷酸化。分子对接研究揭示了化合物12b与COX-2蛋白活性位点的强结合亲和力，这证实了化合物12b作为一种抗炎介质发挥作用。这些结果表明，具有4,5-二氢吡唑噻唑结构的甾体衍生物在设计抗炎药物方面可能值得进一步研究和骨架优化，化合物12b可能是一种有前景的治疗性抗炎药物候选物。
