香豆素-苯丁酸氮芥衍生物的合成及抗口腔肿瘤活性研究  

Synthesis and oral tumor surpress effects of coumarins-chlorambucil derivatives

作　　者：白国辉 张磊 陈彬 王月月 刘霞 艾铝 冉昕 陆明洋 刘建国 赵德刚[5] BAI Guo-hui;ZHANG Lei;CHEN Bin;WANG Yue-yue;LIU Xia;AI Lv;RAN Xin;LU Ming-yang;LIU Jianguo;HAO De-gang(School of Stomatology,Zunyi Medical University,Zunyi 563099,China;Specical Key Laboratory of Oral Diseases Research,Higher Education Insitutions In Guizhou Province,Zunyi 563099,China;School of Pharmacy,Zunyi Medical University,Zunyi 563099,China;Cell Engineering Laboratory,Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;Key laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Institute of Agro-bioengineering,Guizhou University,Guiyang 550025,China)

机构地区：[1]遵义医科大学口腔医学院,贵州遵义563099 [2]贵州省普通高校口腔疾病研究特色重点实验室,贵州遵义563099 [3]遵义医科大学药学院,贵州遵义563099 [4]遵义医科大学附属医院细胞工程实验室,贵州遵义563000 [5]贵州大学农业生物工程研究院,山地植物资源保护与保护种质创新教育部重点实验室,贵州贵阳550025

出　　处：《化学研究与应用》2025年第2期291-301,共11页Chemical Research and Application

摘　　要：本文设计合成了一系列杜仲活性成分香豆素类与苯丁酸氮芥衍生物(B-1~B-4),所有化合物均通过1HNMR、13C-NMR、HRMS进行结构确认。通过CCK-8法筛选出新型衍生物B-2对A-253细胞具有较高的抗增殖作用,IC_(50)值为18.490±1.398μM,且对人正常细胞的毒性最低,IC_(50)值为80.9±10.432μM。采用流式细胞术、蛋白免疫印迹技术、吖啶橙AO染色等方法,研究了B-2对A-253细胞细胞凋亡、细胞周期及线粒体膜电位的影响。研究发现,随着B-2浓度的升高,A-253细胞处于G2/M期的比例升高,凋亡细胞逐渐增多,线粒体功能受损逐渐加重。B-2以浓度依赖的方式下调了细胞周期相关蛋白CDK2和Cyclin A2的表达,使A-253细胞周期停滞;上调细胞凋亡关键蛋白Cleaved caspase-3和Cleaved PARP而诱导A-253细胞发生凋亡;激活自噬相关标记物Beclin 1和LC3的表达,诱导A-253细胞发生自噬,进一步抑制细胞生长。In this paper,a series of novel derivatives of coumarins in eucommia ulmoides and chlorambucil(B-1~B-4)were designed and synthesized,and compounds'structures were identified by~1H-NMR,~(13)C-NMR and HRMS.The new derivative B-2 was selected by CCK-8 method and showed stronger antiproliferative activity against A-253 cells with IC_(50)value of 18.490±1.398μM,and with lower toxicity no normal cells with IC_(50)80.9±10.432μM.The effects of B-2 on apoptosis,cell cycle and mitochondrial membrane potential of A-253 cells was examined by flow cytometry,western blotting and acridine orange AO staining.It was found that after treating A-253 cells with B-2,with the increase of B-2 concentration,the proportion of A-253 cells in G2/M phase,the early apoptotic cells and late apoptotic cells gradually increased,and the impairment of mitochondrial function was gradually aggravated.The B-2 could inhibit the proliferation of A-253 cells through a variety of molecular biological mechanisms,including down-regulation of CDK2 and Cyclin A2 expression to arrest the cell cycle,up-regulation expression of Cleaved Caspase-3 and Cleaved PARP to induce cell apoptosis,raising Beclinl and LC3 expression to induce autophagy.

关 键 词：杜仲 香豆素 苯丁酸氮芥 分子杂合 人口腔鳞状细胞癌 抗癌分子机制 

分 类 号：O626[理学—有机化学]