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目的设计合成一系列去氢表雄酮衍生物,通过对抑制脂多糖(LPS)诱导小胶质细胞激活作用的筛选,以期发现具有抗神经炎症活性的化合物。方法以去氢表雄酮或15β,16β-环丙烷-去氢表雄酮为起始原料,在C-3位β羟基上通过成酯缩合的方法引入天然构型的氨基酸片段,并结合化学合成和生物发酵手段,对C-7位、C-4位进行氧化合成目标化合物;建立了小胶质细胞系BV-2细胞模型,通过LPS的刺激,以抑制一氧化氮(NO)释放量为检测指标,测试化合物的体外抗炎作用,并结合MTT实验检测化合物的细胞毒性。结果与结论通过对相关文献方法的改进和优化,合成了16个目标化合物,其结构经核磁共振氢谱、质谱确证;对小胶质细胞LPS诱导炎症反应抑制作用的活性测试结果表明,在20μmol·L-1时绝大部分目标化合物对一氧化氮的释放有较好的抑制作用,但是具有15β,16β-环丙烷结构的化合物基本都丧失了对NO释放的抑制作用,而具有双羟基结构的目标化合物(9和10)在保持很低的细胞毒性的情况下,对NO的释放抑制作用略有提高,并且呈现剂量依赖性。相关构效关系及作用机制值得进一步研究。
Objective To design and synthesize a series of dehydroepiandrosterone derivatives, and to screen compounds that inhibit the activation of microglia induced by lipopolysaccharide (LPS) in order to find compounds with anti-neuroinflammatory activity. Methods Starting from dehydroepiandrosterone or 15β, 16β-cyclopropane-dehydroepiandrosterone, the naturally-occurring amino acid fragment was introduced into the β-hydroxy group at the C-3 position by ester condensation. Synthesis and biofermentation methods to synthesize target compounds at C-7 and C-4 positions. A microglia cell line BV-2 cell model was established and stimulated by LPS to inhibit the release of nitric oxide (NO) To test the index, the compounds were tested for their anti-inflammatory activity in vitro and the cytotoxicity of the compounds was tested in combination with the MTT assay. RESULTS AND CONCLUSIONS Sixteen target compounds were synthesized by the modification and optimization of related literatures. Their structures were confirmed by 1H-NMR and MS-MS. The inhibitory effect on LPS-induced microglia LPS-induced inflammatory reaction showed that at 20μmol Most of the target compounds at L-1 had a better inhibitory effect on the release of nitric oxide, but the compounds with 15β, 16β-cyclopropane basically lost their inhibitory effect on the release of NO, but had a double hydroxyl structure Of the target compounds (9 and 10) showed a slight increase in inhibition of NO release while maintaining a low cytotoxicity, in a dose-dependent manner. Related structure-activity relationship and mechanism of action worth further study.