菲并咪唑衍生物的体外抗肿瘤活性及其对斑马鱼胚胎发育的毒性效应研究

Evaluation on in vitro anticancer effect against cancer cells and toxicity in vivo of phenanthroimidazole derivatives on zebrafish embryos

目的设计合成菲并咪唑衍生物L271，并研究其外抗肿瘤活性，及其对斑马鱼胚胎发育的毒性效应。方法以1，10-邻菲哕啉-5，6-二酮和2-甲基苯甲醛为原料，运用微波辅助合成技术制备了菲并咪唑衍生L271。采用噻唑蓝（MTT）比色法研究菲并咪唑衍生物L271对人宫颈癌细胞Hela和人肺腺癌细胞A549在体外生长的抑制作用。以斑马鱼为模型，研究L271对斑马鱼胚胎发育的形态、孵化率、死亡率和畸形率的影响，评价L271对斑马鱼胚胎发育的毒性效应。结果经电喷雾质谱技术（ESI-MS）表征确证成功合成了目标化合物L271。新合成的菲并咪唑衍生物L271对人宫颈癌细胞Hela和人肺腺癌细胞A549均有增殖抑制作用，尤其对人宫颈癌细胞Hela的IC50值达到（8．38±0．09）μmol／L，与同等条件下吡柔比星的抗肿瘤活性相当[IC50=（6．97±0．07）μmol／L]。与对照组相比，L271浓度≥15μmol／L暴露组能够引起斑马鱼出现尾鳍萎缩、脊柱弯曲、卵黄囊水肿和心包囊水肿等畸形现象；L271浓度≥30μmol／L可显著降低斑马鱼的孵化率和提高死亡率（P〈0．05）；在48、72、96hpf时，L271对斑马鱼胚胎半致死浓度LC50分别是37．331μmol／L（95％CI：35．535-39．301）、34．911μmol／L（95％CI：33．213-36．729）、30．283μmol／L（95％CI：29．590-30．980k在L271浓度≥15μmol／L时，随着L271浓度的逐渐增加，各暴露组正常胚胎百分率渐下降，胚胎死亡率逐渐上升，而胚胎畸形率先升后降。结论L271对人宫颈癌细胞Hela具有良好的抗肿瘤活性，且与吡柔比星的相当；L271浓度≤10μmol／L对斑马鱼胚胎发育无明显毒性效应。

Objective To design and synthesize phenanthroimidazole derivatives L271, and to evaluate in vitro antitumor activity and toxicity in vivo of phenanthroimidazole derivatives on zebrafish embryos. Methods Using 1,10-phenanthroline-5,6-dione and 2-methylbenzaldehyde as raw material, phenanthroimidazole derives L271 was prepared by microwave-assisted synthesis technology. The inhibition effects of phenanthroimidazole derivatives L271 on the growth of human cervical cancer cells Hela and human hmg adenocarcinoma cells A549 were detected by MTT assay. With zebrafish embryos as a model, the effects of phenanthroimidazole derivatives L271 on the morphology, hatching rates, mortality rates, and malformation rates of zebrafish embryos were investigated to study acute toxicity. Results The chemical structure of target compound L271 was characterized by ESI-MS. Phenanthroimidazole derives L271 showed anti-proliferation effect on human cervical cancer Hela cells and human lung adenocarcinoma A549 cells cultured in vitro, especially for human cervical cancer Hela cells. The inhibitory activity （IC50） of L271 against human cervical cancer Hela cells was about （8.38±0.09）μmol/L, which was close to pirarubicin [IC50 = （6.97±0.07）pmol/L] at the same conditions. Compared with control groups, several abnormalities caused by L271 at 〉 15μmol/L were observed including caudal fin atrophy, spinal curvature, yolk sac edema, pericardial edema, tail bending and etc.. When the concentration of L271 was equal or great than 30μmol/L, hatching rates of zebrafish embryos were reduced significantly, while mortality rates were increased significantly （P〈0.05）. The median lethal concentration （LCs0） of L271 on zebrafish embryos at 48, 72, 96 hpf were 37.331μmol/L（95%CI： 35.535-39.301）, 34.911μmol/L（95%CI： 33.213-36.7290）, 30.283μmol/L（95%CI： 29.590-30.980）, respectively. When the concentration of L271 was equal or great than 15μmol/L, the percentage of normal reduced, and the percentage of mortality increased, while the percentage of mortality increased firstly and then decreased as the concentration of L271 increased. Conclusion L271 exhibit considerable inhibitory effects on human cervical cancer cells Hela, which is comparable to that of pirarubicin. And no obvious toxic effects of L271 on zebrafish embryo when the concentration of L271 is less than 10μmol/L.