目的 设计并合成了一系列羽扇豆醇-3-吡唑侧链衍生物，并考察其体外抗肿瘤活性。方法 以羽扇豆醇为原料，通过取代和酰化反应得到目标化合物4a～o。采用MTT法检测各化合物对HepG2、MCF-7、A549、MRC-5细胞增殖的影响。使用1、2、4 μmol/L化合物4a作用A549细胞48 h后，采用DCFH-DA染色法检测细胞活性氧水平，流式细胞术（AnnexinV-FITC/PI法）检测细胞凋亡水平。分子对接分析化合物4a与HSP90蛋白的结合模式；Western blotting法检测A549细胞中HSP90及凋亡途径相关蛋白表达水平。结果 合成了15个羽扇豆醇-3-吡唑侧链衍生物。化合物4a对A549细胞具有较强的细胞毒性，以剂量相关方式促进活性氧的产生，通过氧化应激诱导细胞凋亡。化合物4a能够更稳定地结合到HSP90蛋白上。化合物4a显著下调HSP90和B淋巴细胞瘤相关X蛋白（Bax）蛋白的表达，同时上调B淋巴细胞瘤2（Bcl-2）蛋白的表达。结论 羽扇豆醇-3-吡唑侧链衍生物4a能够通过促进活性氧产生，导致细胞氧化应激，并诱导A549细胞凋亡。

Objective To design and synthesize a series of lupeol-3-pyrazole side chain derivatives, and to study their antitumor activities in vitro. Methods Starting from lupeol, the target compounds 4a — o were obtained through substitution and acylation reactions. MTT assay was used to detect the effect of compounds on the proliferation of HepG2, MCF-7, A549, and MRC-5 cells. A549 cells were treated with compound 4a at 1, 2, and 4 μmol/L for 48 h, and the level of reactive oxygen species (ROS) was detected by DCFH-DA staining, and cell apoptosis was detected by flow cytometry (AnnexinV-FITC/PI). The binding mode of compound 4a with HSP90 was analyzed by molecular docking, and Western blotting method was used to detect the expression levels of HSP90 and apoptosis-related proteins in A549 cells. Results 15 Lupeol-3-pyrazole side chain derivatives were synthesized. Compound 4a had a strong cytotoxicity on A549 cells, promoted the production of reactive oxygen species in a dose-dependent manner, induced apoptosis through oxidative stress. Compound 4a had a better binding affinity with HSP90, and significantly down-regulated the protein expression of HSP90 and Bax, but up-regulated the protein expression of Bcl-2. Conclusion Lupeol-3-pyrazole side-chain derivative 4a can promote the production of reactive oxygen species, induce cellular oxidative stress, and induce apoptosis in A549 cells.

R914;R966

齐齐哈尔市科技局联合引导项目（LSFGG-2023026）