Department of Pharmacy, The Affiliated Tumor Hospital of Shantou University Medical College, Shantou 515041, Guangdong, China.
Department of Pharmacology, Shantou University Medical College, Shantou 515041, Guangdong, China.
Steroids. 2021 Jul;171:108830. doi: 10.1016/j.steroids.2021.108830. Epub 2021 Apr 6.
To identify new potential anti-inflammatory agents, we herein report the synthesis of novel steroidal chalcones with 3β-pregnenolone esters of cinnamic acid derivatives using pregnenolone as the starting material. The structures of the newly synthesised compounds were confirmed by H NMR, C NMR, HRMS and infrared imaging. All the derivatives were examined to determine their in vitro anti-inflammatory profiles against LPS-induced inflammation in RAW 264.7 cells; the derivates were evaluated by the quantification of the pro-inflammatory mediator nitric oxide (NO) in the cell culture supernatant based on the Griess reaction, which measures nitrite levels, followed by an in vitro cytotoxicity study. Among these novel derivatives, compound 11e [3β-3-phenyl acrylate-pregn-5-en-17β-yl-3' -(p-fluoro)-phenylprop-2'-en-1'-one] was identified as the most potent anti-inflammatory agent, which showed significant anti-inflammatory activity by inhibiting the LPS-induced pro-inflammatory mediator NO in a dose-dependent manner without any cytotoxicity. Moreover, compound 11e markedly inhibited the expression of pro-inflammatory cytokines, including inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2), in LPS-induced RAW 264.7 cells. Further studies confirmed that compound 11e significantly suppressed the transcriptional activity of NF-κB in activated RAW 264.7 cells. Molecular docking study revealed the strong binding affinity of compound 11e to the active site of the pro-inflammatory proteins, which confirmed that compound 11e acted as an anti-inflammatory mediator. These results indicated that steroidal chalcones with 3β-pregnenolone esters of cinnamic acid derivatives might be considered for further research in the design of anti-inflammatory drugs, and compound 11e might be a promising therapeutic anti-inflammatory drug candidate.
为了寻找新的潜在抗炎剂，我们在此报告了使用孕烯醇酮作为起始原料，用肉桂酸衍生物的 3β-孕烯醇酮酯合成新型甾体查耳酮的方法。新合成的化合物的结构通过 1H NMR、13C NMR、高分辨率质谱和红外成像得到确认。所有衍生物均进行了体外抗炎谱测定，以确定其对 LPS 诱导的 RAW 264.7 细胞炎症的抑制作用；通过格里斯反应（用于测量亚硝酸盐水平）测定细胞培养上清液中促炎介质一氧化氮（NO）的定量来评估衍生物，随后进行体外细胞毒性研究。在这些新型衍生物中，化合物 11e [3β-3-苯基丙烯酰基-孕-5-烯-17β-基-3' -（对氟）-苯基丙-2'-烯-1'-酮]被鉴定为最有效的抗炎剂，它通过抑制 LPS 诱导的促炎介质 NO 的剂量依赖性方式表现出显著的抗炎活性，而没有任何细胞毒性。此外，化合物 11e 明显抑制了 LPS 诱导的 RAW 264.7 细胞中促炎细胞因子，包括诱导型一氧化氮合酶（iNOS）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和环氧化酶-2（COX-2）的表达。进一步的研究证实，化合物 11e 显著抑制了激活的 RAW 264.7 细胞中 NF-κB 的转录活性。分子对接研究表明，化合物 11e 与促炎蛋白的活性位点具有很强的结合亲和力，这证实了化合物 11e 作为抗炎介质的作用。这些结果表明，具有肉桂酸衍生物的 3β-孕烯醇酮酯的甾体查耳酮可能被认为是进一步研究抗炎药物设计的候选物，并且化合物 11e 可能是一种有前途的治疗性抗炎药物候选物。
