目的 設(shè)計(jì)并合成了一系列羽扇豆醇-3-吡唑側(cè)鏈衍生物，并考察其體外抗腫瘤活性。方法 以羽扇豆醇為原料，通過取代和?；磻?yīng)得到目標(biāo)化合物4a～o。采用MTT法檢測(cè)各化合物對(duì)HepG2、MCF-7、A549、MRC-5細(xì)胞增殖的影響。使用1、2、4 μmol/L化合物4a作用A549細(xì)胞48 h后，采用DCFH-DA染色法檢測(cè)細(xì)胞活性氧水平，流式細(xì)胞術(shù)（AnnexinV-FITC/PI法）檢測(cè)細(xì)胞凋亡水平。分子對(duì)接分析化合物4a與HSP90蛋白的結(jié)合模式；Western blotting法檢測(cè)A549細(xì)胞中HSP90及凋亡途徑相關(guān)蛋白表達(dá)水平。結(jié)果 合成了15個(gè)羽扇豆醇-3-吡唑側(cè)鏈衍生物。化合物4a對(duì)A549細(xì)胞具有較強(qiáng)的細(xì)胞毒性，以劑量相關(guān)方式促進(jìn)活性氧的產(chǎn)生，通過氧化應(yīng)激誘導(dǎo)細(xì)胞凋亡。化合物4a能夠更穩(wěn)定地結(jié)合到HSP90蛋白上?；衔?b>4a顯著下調(diào)HSP90和B淋巴細(xì)胞瘤相關(guān)X蛋白（Bax）蛋白的表達(dá)，同時(shí)上調(diào)B淋巴細(xì)胞瘤2（Bcl-2）蛋白的表達(dá)。結(jié)論 羽扇豆醇-3-吡唑側(cè)鏈衍生物4a能夠通過促進(jìn)活性氧產(chǎn)生，導(dǎo)致細(xì)胞氧化應(yīng)激，并誘導(dǎo)A549細(xì)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

齊齊哈爾市科技局聯(lián)合引導(dǎo)項(xiàng)目（LSFGG-2023026）