13-取代小檗碱衍生物的合成及其抗肿瘤活性研究

Synthesis and structure-activity relationship of 13-substituted berberine derivatives as anti-cancer agents

目的通过在小檗碱的13位上引入不同取代基,探讨13-取代小檗碱衍生物的体外抗肿瘤活性构效关系,并对其抗肿瘤作用机制开展初步研究。方法以小檗碱为起始原料,13-烷基小檗碱(3a-3j)可经选择性还原后与脂肪醛反应而制备;13-取代苄基小檗碱(3k-3n)可经碱性条件下丙酮加成后与取代溴苄反应而制备。采用磺酰罗丹明B(SRB)法和流式细胞仪分别检测化合物的肿瘤细胞活性及周期的影响。结果设计合成了14个目标化合物,其中7个为全新化合物,其结构经1H-NMR和高分辨质谱确证。所有目标物的抗肿瘤活性均强于小檗碱,其中3h对人肝癌HepG2细胞和白血病细胞HL60的IC50分别达到了0.08μg/mL和0.06μg/mL。初步构效关系研究表明:13位烷基取代衍生物活性明显优于13位苄基取代衍生物;13位烷基链长度适中时活性最好。初步作用机制显示:3h可将肿瘤细胞周期阻滞于G2/M期,使细胞增殖受阻,达到抗肿瘤效果。结论 13-取代小檗碱具有发展成为一类新型抗肿瘤药物的潜力。

Objective To investigate the structure-activity relationship of anti-tumor activities and the corresponding mechanism of 13-substituted berberine derivatives.Methods Berberine was used as the starting material,13-alkyl berberine derivatives were gained by a selective reduction followed by the reaction with fatty aldehyde,while 13-ben zyl berberine derivatives were prepared by an additional reaction with acetone in basic condition followed by the reac tion with various substituted benzyl bromides.SRB assay was used to detect anti-tumor activities and the flow cytome try assay was used to analyze the effect on cell cycle distribution induced by the compounds.Results 14 target compounds were designed and synthesized in this study,7 of them were novel compounds,and their structures were deter mined by 1H-NMR and high resolution mass（HRS）.All the target compounds showed higher anti-tumor activities than berberine,and especially,compound 3h exhibited the potential anti-cancer activities,on HepG2 and HL60 cells with IC50 values of 0.08 and 0.06 μg/mL,respectively.The primary structure-activity relationship demonstrated that 13 alkyl berberine derivatives showed obviously higher anti-tumor activities than 13-benzyl berberine derivatives,and a moderate length of the alkyl chain was beneficial for the activity.The primary mechanism study showed that cell cycle blockage at G2/M phase in HCT116.Conclusion 13-substituted berberine derivatives are promising as a kind of novel anti-tumor agents.