钩吻生物碱衍生物的设计、合成和抗结直肠癌活性研究

Design,Synthesis and Anti-colon Cancer Activity of Gelsemium Elegans Alkaloid Derivatives

目的:围绕有毒中药钩吻生物碱抗肿瘤药效骨架,引入经典抗肿瘤药效基团,设计并合成新型钩吻生物碱衍生物,评价其对结直肠癌细胞株体内外抗肿瘤活性。方法:通过药效结构拼合的方法,以钩吻生物碱共同药效骨架C3-氧化吲哚螺环结构为出发点,引入抗肿瘤药效基团异噁唑片段,设计目标化合物;以含氧化吲哚骨架的MBH碳酸酯和3-甲基-4-硝基-5-烯基异噁唑为原料,通过高度区域选择性1,6-环加成反应,制得新型钩吻生物碱衍生物——含有异噁唑基团的3-螺环戊烯-2-氧化吲哚;用cell counting Kit-8(CCK-8)法测定新型钩吻生物碱衍生物对多种人结直肠癌肿瘤细胞株的增殖抑制作用;用人结直肠癌细胞HCT116采用皮下注射法构建小鼠肿瘤模型,观察不同浓度钩吻生物碱衍生物的体内抗肿瘤作用。结果:得到产率为84%的目标钩吻生物碱衍生物(3),新化合物结构经~1H-NMR,^(13)C-NMR和HRMS确证,为甲基-5′-氯-4-(4-异丙基苯基)-5-(3-甲基-4-硝基异噁唑-5-基)-2′-氧代-螺[环戊烷-1,3′-吲哚]-2-烯-2-羧酸酯;该新型钩吻生物碱衍生物能抑制LS180、HT-29、 DLD-1、HCT116、SW620、SW480和SW1116细胞增殖,其半数抑制浓度(IC_(50))分别为0.69,1.26,2.26,0.38,3.26,4.98和2.57 mg/L,其中对HCT116的抑制作用较阳性对照药物阿霉素更好;钩吻生物碱衍生物高、中、低剂量组腹腔注射给药均能抑制HCT116模型小鼠肿瘤生长,其体内抗肿瘤生物活性呈剂量依赖。结论:该新型钩吻生物碱衍生物具有良好的抗结直肠癌活性,可作为抗肿瘤先导药物进行进一步研究,拓展创新中药途径。

Objective:Based on the anti-tumor efficacy scaffold of alkaloids from a toxic traditional Chinese medicine,Gelsemium elegans,design and synthesize novel Gelsemium elegans alkaloid derivatives by introducing classic anti-tumor efficacy groups,and evaluate their in vivo and in vitro anti-tumor activity against colon cancer cell lines.Methods:Using a hybridization approach,the common pharmacophore scaffold of Gelsemium elegans alkaloid,C3-oxindole spirocyclic scaffold,was used as a starting point,and the anti-tumor efficacy group,isoxazole fragment,was introduced to design the target compound.Using oxindole-derived MBH carbonates and 3-methyl-4-nitro-5-alkenylisoxazoles as starting materials,a highly regioselective 1,6-cycloaddition reaction was applied to obtain a novel Gelsemium elegans alkaloid derivative containing an isoxazole group.The Cell Counting Kit-8(CCK-8)method was used to determine the proliferation inhibitory effects of the novel compound on various human colon cancer cell lines.HCT116 human colon cancer cells were used to construct a mouse tumor model via subcutaneous injection,and the in vivo anti-tumor effects of different concentrations of the Gelsemium elegans alkaloid derivative were observed.Results:A target Gelsemium elegans alkaloid derivative was obtained with a yield of 84%,and the structure was confirmed by 1H-NMR,13 C-NMR,and HRMS.The Gelsemium elegans alkaloid derivative could inhibit the proliferation of LS180,HT-29,DLD-1,HCT116,SW620,SW480,and SW1116 cells,with IC 50 values of 0.69,1.26,2.26,0.38,3.26,4.98,and 2.57 mg/L,respectively.The inhibitory effect on HCT116 was better than that of the positive control drug mitomycin.The Gelsemium elegans alkaloid derivative in high,medium,and low dose groups could inhibit the growth of HCT116 model mouse tumors when administered by intraperitoneal injection,and the in vivo anti-tumor biological activity was dose-dependent.Conclusion:The novel Gelsemium elegans alkaloid derivative has good anti-colon cancer activity and can be further researched as a lead anti-tumor drug to improve traditional Chinese medicine innovation capabilities.