β-榄香烯取代哌嗪酰胺类衍生物的合成及抗肿瘤活性研究

Synthesis and anti-proliferative activity of β-elemene substituted piperazine amide derivatives

目的设计合成β-榄香烯取代哌嗪酰胺类衍生物并进行体外抗癌活性筛选。方法通过合成β-榄香烯单氯代物,在其结构中引入取代哌嗪结构来合成β-榄香烯取代哌嗪衍生物,然后再与取代苯甲酰氯或取代苯丙烯酰氯反应制得β-榄香烯取代哌嗪酰胺类衍生物。采用MTT法测定了目标化合物对人宫颈癌细胞、人肝癌细胞、人纤维肉瘤细胞等10种细胞的增殖抑制作用。结果与结论合成了23个未见文献报道的β-榄香烯取代哌嗪酰胺类衍生物。经1H-NMR、MS波谱分析确证结构。其中21个化合物经MTT法测定了体外抗肿瘤活性,结果显示活性大多高于β-榄香烯。

Elemene emulsion is a broad spectrum antitumor drug, whose main component is β-elemene. For improving its solubility and increasing its antitumor activity,β-elemene had been structurally modified and about three hundreds β-elemene derivatives were synthesized and evaluated for their proliferation-restraining activity in vitro. Among them,β-elemene substituted piperazine derivatives DX-1 and DX-2 show higher an- titumor activities both in vitro and in vivo than β-elemene. Mechanism studies suggested the apoptosis induc- tion ability of DX-1 was associated with the generation of hydrogen peroxide（ H202 ）, a decrease of mito- chondrial membrane potential（MMP） ,and the activation of caspase-8. The activation of caspase-8 by DX-1 was correlated with the decrease in the levels of cellular FLICE-inhibitory protein （c-FLIP）. Furthermore DX-1 and DX-2 could inhibit cell growth through mTOR and/or AKT activity inhibition. To improve higher toxicities of DX-1 and DX-2 in vivo,β-elemene substituted piperazine amide derivatives were designed and synthesized from β-elemene via chlorination and nucleophilic substitution. That is,β-elemene was firstly re- acted with sodium hypochlorite （NaC10） to generate β-elemene chlorides and the chlorides of β-elemene were then condensed with substituted piperazine, sequentially β-elemene substituted piperazine derivatives were acylated with substituted benzoyl chloride or benzoacryloyl chloride. The products were separated with column chromatography. The structures of the target compounds were confirmed by 1H-NMR and MS. The in vitro anti-proliferative effects of the target compounds on 10 species of tumor cells such as HeLa, HepG2, HT-1080, etc. were determined by MTT assay. Preliminary pharmacological results showed that most com- pounds possessed more potent anti-proliferative activity than β-elemene and some compounds were lower than β-elemene substituted piperazine derivatives ,but the substituted piperazine amide derivatives had better selectivity on tumor cell, such as stronger inhibitory activity on human melanoma cell（A375-S2）.