摘要
目的 设计并合成新型苯并咪唑衍生物,并对其体外抗肿瘤活性进行评价。方法 以5-氯-2-硝基苯胺为原料,经碘代、亲核取代、Heck偶联和缩合反应得到目标化合物。采用噻唑蓝(MTT)法评价目标化合物对肺癌A549、肝癌HepG-2和宫颈癌Ca-Ski三种肿瘤细胞的增殖抑制活性以及对人正常肝组织LO-2细胞及肾上皮HEK-293T细胞的毒性,初选抗肿瘤活性较好的化合物;采用生物膜干涉技术分析优选化合物与组蛋白去乙酰化酶8(HDAC8)之间的亲和作用;采用分子对接探究优选化合物与HDAC8的分子对接模式;采用蛋白免疫印迹法研究优选化合物对HDAC8蛋白的抑制作用;采用酶抑制实验检测优选化合物对HDAC8的抑制作用;采用血浆稳定性实验考察优选化合物的稳定性。结果与结论合成了10个全新的苯并咪唑衍生物,其结构均经ESI-HRMS、1H-NMR和13C-NMR谱确证。抗肿瘤活性和细胞毒性实验筛选出对肝癌HepG-2细胞抑制活性较好的化合物14b,其IC50值为(2.68±0.68)μmol·L^(-1);生物膜干涉技术分析结果显示,化合物14b与HDAC8的亲和力常数为8.31×10-6mol·L^(-1),表明其与HDAC8有较强的亲和能力;分子对接结合模式分析显示,化合物14b中异羟肟酸基团与HDAC8蛋白(PDB ID:6HQY)关键氨基酸His142形成氢键作用,并与锌离子螯合;蛋白免疫印迹实验结果显示,化合物14b在8μmol·L^(-1)浓度时能有效抑制HDAC8蛋白的表达;酶抑制实验结果显示,化合物14b对HDAC8的IC50值为0.75μmol·L^(-1),表明其对HDAC8有一定的抑制作用;SD大鼠血浆稳定性实验结果显示,化合物14b半衰期为1 h,表明其在大鼠血浆中的稳定性有待提高。化合物14b可能通过与HDAC8结合从而发挥抗肝癌细胞增殖作用,具有进一步研究的价值。
Based on the previous work of the research group and the principle of combinatorial chemistry,a benzene ring was utilized as the linker to connect the benzimidazole moiety with the hydroxamic acid.The steric hindrance provided by the benzene ring is hypothesized to enhance HDAC8 inhibitory activity.Ten 6-substituted-2-aryl benzimidazole derivatives were synthesized by introducing diverse acryloyl groups at the 6-position of benzimidazole or retaining hydrogen at the 6-position.The structures of the target compounds were characterized by ESI-HRMS,^(1)H-NMR and ^(13)C-NMR.The synthesized compounds were subsequently evaluated for in vitro antitumor activity and target validation.The in vitro antitumor activity results demonstrated that compound 14b exhibited potent activity against HepG-2 cells with an IC_(50) value of(2.68±0.68)μmol·L^(-1).Western blot analysis revealed that compound 14b suppressed expression of HDAC8 protein.Kinase inhibition assays further indicated that compound 14b displayed inhibitory effect on HDAC8 with an ICso value of 0.74μmol·L^(-1).Additionally,bio-layer interferometry results confirmed the strong binding affinity between compound 14b and HDAC8.
作者
陈俊杰
王伟旭
张伟怡
姚军
马成
CHEN Junje;WANG Weixu;ZHANG Weiyi;YAO Jun;MA Cheng(School of Pharmacy,Xinjiang Medical University,Urumqi 830011,China;Xinjiang Key Laboratory of Natural Medicines Active Components and Drug Release Technology,Xinjiang Medical University,Urumqi 830011,China;Xinjiang Key Laboratory of Biopharmaceuticals and Medical Devices,Xinjiang Medical University,Urumqi 830011,China;Engineering Research Center of Xinjiang and Central Asian Medicine Resources,Xinjiang Medical University,Urumqi 830011,China;State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,Xinjiang Medical University,Urumqi 830011,China)
出处
《中国药物化学杂志》
2025年第3期167-179,共13页
Chinese Journal of Medicinal Chemistry
基金
中央引导地方科技发展专项资助项目(ZYYD2025JD12)
新疆天然药物活性组分与释药技术重点实验室资助项目(2024XJTRZ01)
国家自然科学基金资助项目(82204438)。
