新型鬼臼毒素衍生物的合成及抗肿瘤活性研究

目的通过对鬼臼毒素进行结构改造,寻找高效低毒的抗肿瘤先导化合物。方法运用拼合原理,将川芎嗪和天冬氨酸等结构片段引入鬼臼毒素结构中,通过1 H-NMR、13 C-NMR和HRMS确证结构;采用四甲基偶氮唑蓝(MTT)法评价化合物对人肺癌细胞A549、人肝癌细胞HepG2和人乳腺癌细胞MCF-7的活性及对人正常肝细胞L-02的毒性,并采用4′,6-二脒基-2-苯基吲哚(DAPI)染色法观察优势化合物对细胞核形态的影响。结果得到3个新型鬼臼毒素衍生物,其中化合物P-02(IC 50=0.1998±0.08μmol·L^-1)对A549的抑制作用与阳性药阿霉素相当(IC 50=0.1881±0.06μmol·L^-1),较依托泊苷(IC 50=8.55±0.43μmol·L^-1)提高了40倍;该化合物对L-02的毒性(IC 50=0.6701±0.176μmol·L^-1)明显小于阿霉素(IC 50=0.04684±0.02μmol·L^-1);DAPI染色结果表明,化合物P-02通过诱发A549细胞核碎裂产生细胞毒性。结论与临床化疗药物依托泊苷和阿霉素相比,将川芎嗪和氨基酸片段引入鬼臼毒素后可增强抗肿瘤活性,降低毒性,为新型鬼臼毒素类先导化合物的发现提供了参考。

Synthesis and evaluation of podophyllotoxin derivatives as the novel anticancer agents

Objective To search for the anti-tumor drug candidates with higher efficiency and lower toxicity by making structural modification of podophyllotoxin.Methods Ligustrazine and aspartic acid were introduced into podophyllotoxin by the principle of combination.The structures of the new compounds were confirmed by 1 H-NMR,13 C-NMR and HRMS analysis.The antitumor activities were tested against A549(human lung cancer)cell line,HepG2(human hepatoma)cell line and MCF-7(human breast cancer)cell line and the cytotoxicity was tested against L-02(human normal liver)cell line by MTT method.4′,6-diamidino-2-phenylindole(DAPI)staining was used to observe the effect of dominant compounds on nuclear morphology.Results Three new podophyllotoxin derivatives were obtained.The antitumor activity of compound P-02(IC 50=0.1998±0.08μmol·L^-1)against A549 cell line was 40 times stronger than the positive drug etoposide(IC 50=8.55±0.43μmol·L^-1),and was comparable to doxorubicin(IC 50=0.1881±0.06μmol·L^-1),moreover the toxicity to L-02(IC 50=0.6701±0.176μmol·L^-1)cell line was significantly less than that of doxorubicin(IC 50=0.04684±0.02μmol·L^-1).Nuclear fragmentation was observed in P-02 treated A549 cells by DAPI staining.Conclusion Compared with the clinical chemotherapy drugs etoposide and doxorubicin,introducing the fragment of ligustrazine and amino acid into podophyllotoxin can enhance the anti-tumor activity and reduce the toxicity.The research result might provide a reference for the discovery of the novel podophyllotoxin derivatives.