摘 要 研究表明，50%以上原癌基因编码产物都具有酪氨酸激酶活性，特别是EGFR-TK在多种人恶性肿瘤细胞中均有过度表达。因此，可通过干扰、阻断EGFR-TK受体信号传到，抑制肿瘤细胞的生长、增殖、迁移等，从而达到肿瘤靶向治疗的目的。喹唑啉结构是一种可抑制EGFR-TK且具有显著的抗肿瘤活性基团，具有较大的研发价值和临床应用前景。鉴于此，本课题拟以喹唑啉为母体，并对其进行合理的结构修饰以期得到新型小分子靶向抗肿瘤药物。具体研究工作介绍如下 1．以喹唑啉作为优势结构骨架，在保持喹唑啉母环及6-位的取代基保持不变，7-位的取代基改变的前提下，同时对其4-位苯氨基进行侧链修饰改造，设计并合成了18个A系列和15个B系列共计33个两类新型的喹唑啉类衍生物，对其进行了1H NMR、13C NMR、MS结构表征。 2．利用MTT法以PC-3、HepG2、A549、MCF-7、DU145、SH-SY5Y六种肿瘤细胞为实验对象对合成的33个新型喹唑啉衍生物进行抗肿瘤活性初步筛选；MTT法结果表明A系列化合物大部分具有较高的体外抑制肿瘤活性，该系列化合物对常见的PC-3、HepG2、A549、MCF-7、DU145、SH-SY5Y六种肿瘤细胞的IC50值均低于10 μM，具有一定的广谱抗癌活性；B系列化合物部分化合物具有较强的选择性抑制活性。 4．对化合物 A-16进行了系统的体内外抗肿瘤活性评价，实验结果表明该化合物对PC-3和DU145细胞均具有显著的选择性抑制活性，IC50分别为3.839 μM和8.093 μM。化合物A-16对于两种前列腺癌细胞体内外活性研究实验表明，该化合物作用于此两种肿瘤细胞后，细胞膜通透性增大，细胞凋亡率显著增加；经JC-1染色后激光共聚焦显微镜观察检测表明该化合物作用于肿瘤细胞后，细胞线粒体膜电位降低，细胞发生凋亡；经荧光光谱仪检测，细胞内活性氧（ROS）水平升高，继而诱导细胞凋亡。分别建立两种肿瘤细胞的荷裸鼠模型，进行化合物A-16的体内抑制肿瘤活性实验结果表明，化合物A-16能够有效抑制体内肿瘤生长和延长裸鼠生存时间，裸鼠体重无明显下降趋势，表明该化合物毒性较低，具有一定的研究价值。 本课题研究工作表明，以喹唑啉作为优势结构骨架,通过改变其6、7位取代基及4-取代基的改变可以得到抗肿瘤活性较好的新化合物。该类化合物作用于PC-3、HepG2、A549、MCF-7、DU145和SH-SY5Y细胞，可以或强或弱地抑制细胞增殖。本研究能够为喹唑啉类化合物在抗肿瘤药物领域的应用提供一定的学术及实验研究依据。

ABSTRACT Studies have shown that more than 50% of the protooncogene encoded products have tyrosine kinase activity, especially EGFR-TK is overexpressed in a variety of human malignant tumor cells. Therefore, it can inhibit the growth, proliferation and migration of tumor cells by interfering with and blocking the signal transmission of EGFR-TK receptors, thereby achieving the purpose of tumor targeted therapy. The quinazoline structure is a kind of EGFR-TK, which has significant anti-tumor activity groups, and has great research and development value and clinical application prospects. In view of this, this study intends to use quinazoline as the mother body and carry out reasonable structural modification to obtain a new small molecule targeted anti-tumor drug. The specific research work is as follows 1. The quinazoline was used as the main structural skeleton, and the side chain modification of the 4-position anilino group was carried out while maintaining the quinazoline parent ring and the 6-position substituent remained unchanged and the 7-position substituent was changed. A total of 33 two new classes of quinazoline derivatives were modified, designed and synthesized in 18 A series and 15 B series. The structures were characterized by 1H NMR, 13C NMR and MS. 2. MTT assay was used to screen the 33 new quinazoline derivatives of PC-3, HepG2, A549, MCF-7, DU145 and SH-SY5Y for the anti-tumor activity. The MTT method showed A series. Most of the compounds have high inhibitory activity in vitro. The IC50 values of the six compounds of PC-3, HepG2, A549, MCF-7, DU145 and SH-SY5Y are lower than 10 μM. Spectrum anticancer activity; some compounds of B series compounds have strong selective inhibitory activity. 3. Compound A-16 was evaluated for its in vitro and in vivo antitumor activity. The results showed that it had strong selective inhibitory activity against PC-3 and DU145 cells, with IC50 of 3.839 μM and 8.093 μM, respectively. Further, in vitro and in vivo activities of compound A-16 on two kinds of prostate cancer cells were carried out. In vitro experiments showed that after the compound acts on the two kinds of tumor cells, the cell membrane permeability is increased and the apoptosis rate is significantly increased; After -1 staining, the results of laser confocal microscopy showed that the mitochondrial membrane potential decreased and the cells undergo apoptosis after the action of the compound. The level of reactive oxygen species (ROS) in the cells was increased by fluorescence spectrometry, which in turn induced apoptosis. The nude mice model of the two tumor cells was established, and the in vivo inhibitory activity of the compound A-16 was tested. The results showed that the compound A-16 can effectively inhibit the tumor growth in vivo, and the body weight of the nude mice did not decrease significantly, indicating that the compound was more toxic. It is low and can effectively prolong the survival time of nude mice, which has certain research value. The research work of this subject showed that Icotinib, Vandetanib, Cantintinib and Lapatinib were used as lead compounds, and new compounds with better antitumor activity could be obtained by changing the substituents at the 6th and 7th positions and the 4-substituent. These compounds act on PC-3, HepG2, A549, MCF-7, DU145 and SH-SY5Y cells and can inhibit cell proliferation either strongly or weakly. This study can provide some academic and experimental research basis for the application of quinazoline compounds in the field of anti-tumor drugs.