论文总字数：31356字

摘 要

在众多的抗肿瘤药物中，顺铂因为抗癌谱广，细胞毒性强的原因，自上市以来一直处于非小细胞肺癌等多种癌症的化疗一线药物。但是由于顺铂有较强的器官毒性和耐药性，削弱了顺铂的抗肿瘤活性和治疗癌症的效果，因此，限制了顺铂在临床上的应用。

通过前期的研究，我们课题组已经合成出了一种毒性更小但有类似于顺铂的抗肿瘤活性的二羧酸配体的卡铂类似物化合物DN604。此外，通过文献的查阅，发现了在多种癌症中都具有明显的GST抑制效果的GST抑制剂NBDHEX，文献证明GST抑制剂会减弱耐药性。

在本次毕设课题中，我们通过多步有机化学的反应合成了化合物DN604-NBDHEX，并对这个化合物进行了¹ H和¹³ C NMR光谱以及ESI-MS光谱仪的结构表征，验证了化合物的结构。我们通过细胞培养，建立了A549，A549/DDP和LO2细胞系，并进行了MTT细胞毒性实验，细胞凋亡实验，细胞周期实验和免疫荧光实验。通过以上的各种实验，证明了DN604-NBDHEX复合物具有优良的抗耐药性和细胞毒性，抗肿瘤活性高，具有进一步研究和应用的价值和潜力。在后续需要通过更多的实验来补充研究其抗肿瘤机理和抗耐药机理。

关键词：抗肿瘤、GST抑制剂、DN604、耐药

Abstract

Among the numerous anti-tumor drugs, cisplatin has been the first-line chemotherapy drug for various cancers such as non-small cell lung cancer since its introduction because of its broad anticancer spectrum and strong cytotoxicity. However, due to the strong organ toxicity and drug resistance of cisplatin, the anti-tumor activity of cisplatin and the effect of treating cancer are weakened, thus limiting the clinical application of cisplatin.

Through previous studies, our group has synthesized a carboplatin analog compound DN604 which is less toxic but has a dicarboxylic acid ligand similar to the antitumor activity of cisplatin. In addition, the GST inhibitor NBDHEX, which has significant GST inhibitory effects in various cancers, was found by literature review, and the literature demonstrates that GST inhibitors attenuate drug resistance.

In this project, we synthesized the compound DN604-NBDHEX by multi-step organic chemistry, and carried out 1 H and 13 C NMR spectroscopy and structural characterization of the ESI-MS spectrometer to verify the structure of the compound. . We established A549, A549/DDP and LO2 cell lines by cell culture, and performed MTT cytotoxicity experiments, apoptosis experiments and cell cycle experiments. Through the above various experiments, it was proved that DN604-NBDHEX complex has excellent anti-drug resistance and cytotoxicity, high anti-tumor activity, and has the value and potential for further research and application. In the follow-up, more experiments are needed to supplement the anti-tumor mechanism and anti-drug mechanism.

KEY WORDS: anti-tumor, GST inhibitor, DN604, drug resistance

目 录

摘 要 Ⅰ

Abstract Ⅱ

第一章 绪论 1

1.1顺铂治疗肺癌 1

1.1.1铂类抗肿瘤药物概述 1

1.1.2新型铂类化合物的研究 2

1.1.3顺铂治疗非小细胞肺癌现状 3

1.2 GST抑制剂 4

1.2.1 GSH、GST与铂类药物耐药性的关联 4

1.2.2 GST抑制剂与顺铂耐药的关联 5

1.3课题设计依据及其意义 7

第二章 实验部分 8

2.1研究目标 8

2.2研究内容 8

2.3具体实验方案——DN604-NBDHEX的合成 8

2.3.1材料和试剂 8

2.3.2 DN604的四价铂前药的合成 8

2.3.3 GST抑制剂NBDHEX的合成 10

2.3.4 DN604-NBDHEX的合成 10

2.4具体实验方案——体外细胞实验 11

2.4.1材料和试剂 11

2.4.2建立细胞系和细胞培养 11

2.4.3 MTT分析 12

2.4.4细胞凋亡实验 12

2.4.5细胞周期实验 12

2.4.6免疫荧光实验 12

第三章 结果与讨论 14

3.1 DN604-NBDHEX的合成 13

3.2体外细胞实验 17

3.2.1建立细胞系和细胞培养 17

3.2.2 MTT分析 17

3.2.3细胞凋亡实验 19

3.2.4细胞周期实验 22

3.2.5细胞免疫荧光实验 25

3.3结论 26

3.4本项目总结与展望 27

缩略语 30

参考文献 31

致 谢 33

第一章 绪论

1.1顺铂治疗肺癌

1.1.1铂类抗肿瘤药物概述

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