臺灣博碩士論文加值系統

根據衛生福利部於公布之最新統計資料(106年)，卵巢癌佔居女性十大癌症排行榜中之第七位。因為目前沒有直接的篩檢手段可以進行檢測，許多民眾亦無定期健檢的習慣，故大部分病患都是等到出現生理上的病症才就醫，而屆時通常已經是治癒率僅剩5%~10%的第四期了。即使在早期的健檢中幸運發現病狀並給予治療，在康復後仍然有高達約八成的復發率，且卵巢癌細胞的抗藥性已於許多研究中發現，故開發一個對於卵巢癌有效的新型藥物是至關重要的。
在本篇研究中，我們藉由已知藥物YM155和順鉑作為控制組，並利用有機合成的方式製備出4種分別有不同官能基團的單倍與雙倍蒽醌咪唑衍生物來做為對照組，再與正常細胞和分別有無抗藥性的兩種卵巢癌細胞進行共孵育來測試其對於細胞之毒性。利用MTT吸光度結果我們得知在本研究中製備的四種化合物中，與YM155有最相似結構特性的ANQI-YM155有最低的IC50，且結果在癌細胞與正常細胞的數值比較均差有數百倍之差距，指示出相較於正常細胞，ANQI系列藥物在癌細胞中有更好的選擇性。根據本次研究結果中可以看到，所有ANQI系列的藥物之IC50均遠高於YM155但低於順鉑，指示出在引導細胞進行凋亡程序時的途徑可能與順鉑相似，即透過影響拓樸異構酶。在臨床上已出現抗藥性的卵巢癌中，本篇研究透過提供了新式的藥物做為未來癌症新藥的潛力。

According to the latest statistical data provided by Health Promotion Administration in 2020, ovarian cancer ranks seventh among the top ten cancers in women. Since there is no directly diagnostic method for ovarian cancer, and most of the people do not have the habit to do the regular health check, so most of the patient only see doctors when they have certain symptoms, but, by then, the state of cancer usually has proceed to the end stage and which is difficult to cure. Even if patients are found and treated during the early stage, there is still a recurrence rate of about 80% after recovery. Moreover, the drug resistance of ovarian cancer cells has been discovered in many studies, so it is crucial to develop a new drug that is effective for ovarian cancer.
In this study, we prepared the known compound YM155 and Cisplatin as control group, and by organic synthesize 4 types of anthraquinone imidazole derivatives as experiment groups for anticancer drug study. Incubating each compounds with normal cells and two ovarian cancer cells with or without drug resistance, to test their toxicity to cells. By using MTT analysis, we know that among the four compounds prepared in this study, ANQI-YM155, which has the most similar structural characteristics to YM155, has the lowest IC50, which has hundreds times different from YM155 in normal cells, suggest the series of ANQI drugs have better selectivity in cancer cells.
According to the results, the IC50 of all the ANQI series of drugs is much higher than YM155 but lower than cisplatin, indicating that the pathway in guiding cells to undergo apoptosis procedures may be similar to that of cisplatin, that is, by acting as topoisomerase. In the clinically resistant ovarian cancer, this research provides the potential of new anticancer drugs in the future.

摘要 i
Abstract ii
致謝 iv
目錄 vi
圖目錄 ix
表目錄 x
附圖目錄 xi
第一章 前言 1
第二章 文獻回顧 2
2.1 卵巢癌 2
2.2 化學治療法 2
2.2.1 酵素抑制劑 3
2.2.2 拓樸酶抑制劑 3
2.2.2.2.1 共價式結合類 5
2.2.2.2.2 非共價式結合類 6
2.3 化合物結構設計 8
2.3.1 YM155 (Sepantronium Bromide) 抑制劑 8
2.3.2 萘醌咪唑衍生物 (Naphthoquinone Imidazolium derivatives) 9
2.3.4 蒽醌類化療藥物 10
2.3.5 咪唑類(Imidazole)藥物 11
2.3.6 蒽醌咪唑衍生物 (Anthraquinone Imidazolium Derivatives)藥物設計開發 11
第三章 實驗目的與策略 13
3.1 實驗目的 13
3.2 實驗策略 13
第四章 實驗方法與藥品材料 15
4.1 實驗藥品 15
4.2 實驗儀器 17
4.3 藥物合成步驟 19
4.3.1 ANQI-NH合成方法 19
4.3.2 ANQI-Acetyl 合成方法 20
4.3.3 ANQI-NH-sEG 合成方法 21
4.3.4 ANQI-sEG 合成方法 22
4.3.5 ANQI-NH-YM155 合成方法 23
4.3.6 ANQI-YM155 合成方法 24
4.3.7 ANQI-NH-EG 合成方法 25
4.3.8 ANQI-EG 合成方法 26
4.3.9 ANQI-NH-Hex 合成方法 27
4.3.10 ANQI-Hex 合成方法 28
4.4 Calf thymus DNA 溶液配置 29
4.5 螢光淬滅常數 29
4.6 細胞培養(Cell culture) 31
4.6.1 細胞解凍 31
4.6.2 細胞復養 31
4.6.3 細胞計數 31
4.7 細胞生存率分析 32
第五章　結果與討論 33
5.1 藥物合成 33
5.1.1 合成機理 33
5.1.2 藥物命名 35
5.2 藥物命名 36
5.3 淬滅常數之結合特性分析 37
5.4 細胞生存率分析 40
第六章　結論 42
參考文獻 43
附錄 48

1.衛生福利部. 衛生福利部公布癌症發生資料. [Internet] 2020 Sep 21 [cited 2020 Jun 6]; Available from: https://www.hpa.gov.tw/Pages/Detail.aspx?nodeid=4141&pid=12682.
2.Agarwal, R. and S.B. Kaye, Ovarian cancer: strategies for overcoming resistance to chemotherapy. Nature Reviews Cancer, 2003. 3(7): p. 502-516.
3.Institute, N.C. Ovarian, Fallopian Tube, and Primary Peritoneal Cancer Prevention (PDQ®)–Patient Version. [Internet] Apr 21; Available from: https://www.cancer.gov/types/ovarian/patient/ovarian-prevention-pdq#section/all.
4.Köberle, B., et al., Cisplatin resistance: preclinical findings and clinical implications. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 2010. 1806(2): p. 172-182.
5.Galluzzi, L., et al., Molecular mechanisms of cisplatin resistance. Oncogene, 2012. 31(15): p. 1869-1883.
6.Thatcher, N., et al., Gefitinib plus best supportive care in previously treated patients with refractory advanced non-small-cell lung cancer: results from a randomised, placebo-controlled, multicentre study (Iressa Survival Evaluation in Lung Cancer). The Lancet, 2005. 366(9496): p. 1527-1537.
7.Brown, J.S., et al., Targeting DNA repair in cancer: beyond PARP inhibitors. Cancer discovery, 2017. 7(1): p. 20-37.
8.Vanhoefer, U., et al., Irinotecan in the treatment of colorectal cancer: clinical overview. Journal of clinical oncology, 2001. 19(5): p. 1501-1518.
9.LOEHRER, P.J. and L.H. EINHORN, Cisplatin. Annals of internal medicine, 1984. 100(5): p. 704-713.
10.Harshman, K.D. and P.B. Dervan, Molecular recognition of B-DNA by Hoechst 33258. Nucleic acids research, 1985. 13(13): p. 4825-4835.
11.Lerman, L., Structural considerations in the interaction of DNA and acridines. Journal of molecular biology, 1961. 3(1): p. 18-IN14.
12.Lankelma, J., et al., Doxorubicin gradients in human breast cancer. Clinical cancer research, 1999. 5(7): p. 1703-1707.
13.Shimizu, T., et al., Phase I safety and pharmacokinetic study of YM155, a potent selective survivin inhibitor, in combination with erlotinib in patients with EGFR TKI refractory advanced non-small cell lung cancer. Cancer chemotherapy and pharmacology, 2020. 86(2): p. 211-219.
14.Nakahara, T., et al., YM155, a novel small-molecule survivin suppressant, induces regression of established human hormone-refractory prostate tumor xenografts. Cancer research, 2007. 67(17): p. 8014-8021.
15.Ho, S.-H.S., et al., Antiproliferative, DNA intercalation and redox cycling activities of dioxonaphtho [2, 3-d] imidazolium analogs of YM155: A structure–activity relationship study. European journal of medicinal chemistry, 2015. 104: p. 42-56.
16.Chen, B.-J., et al., Bisintercalating DNA redox reporters for real-time electrochemical qLAMP. Biosensors and Bioelectronics, 2019. 129: p. 277-283.
17.Braña, M.F., et al., Synthesis and cytostatic activity of benz (de) isoquinolin-1, 3-diones. Structure-activity relationships. 1981.
18.Bousquet, P.F., et al., Preclinical evaluation of LU 79553: a novel bis-naphthalimide with potent antitumor activity. Cancer research, 1995. 55(5): p. 1176-1180.
19.Braña, M.F., et al., New analogues of amonafide and elinafide, containing aromatic heterocycles: synthesis, antitumor activity, molecular modeling, and DNA binding properties. Journal of medicinal chemistry, 2004. 47(6): p. 1391-1399.
20.Palchaudhuri, R. and P.J. Hergenrother, DNA as a target for anticancer compounds: methods to determine the mode of binding and the mechanism of action. Current opinion in biotechnology, 2007. 18(6): p. 497-503.
21.Saeed, H.K., et al., The Structure of Linkers Affects the DNA Binding Properties of Tethered Dinuclear Ruthenium (II) Metallo‐Intercalators. Chemistry–A European Journal, 2017. 23(23): p. 5467-5477.
22.Hussain, H., et al., A fruitful decade from 2005 to 2014 for anthraquinone patents. Expert opinion on therapeutic patents, 2015. 25(9): p. 1053-1064.
23.Yaoxian, W., et al., Emodin induces apoptosis of human cervical cancer hela cells via intrinsic mitochondrial and extrinsic death receptor pathway. Cancer cell international, 2013. 13(1): p. 1-8.
24.Min, H., et al., Emodin reverses leukemia multidrug resistance by competitive inhibition and downregulation of P-glycoprotein. PLoS One, 2017. 12(11): p. e0187971.
25.Tang, N., et al., Rhein induces apoptosis and autophagy in human and rat glioma cells and mediates cell differentiation by ERK inhibition. Microbial pathogenesis, 2017. 113: p. 168-175.
26.Chen, R., et al., Potential antineoplastic effects of Aloe-emodin: a comprehensive review. The American journal of Chinese medicine, 2014. 42(02): p. 275-288.
27.Wang, C.C. and P.B. Dervan, Sequence-specific trapping of topoisomerase I by DNA binding polyamide− camptothecin conjugates. Journal of the American Chemical Society, 2001. 123(36): p. 8657-8661.
28.Frank, N.E., et al., Comparative effects of doxorubicin and a doxorubicin analog, 13-deoxy, 5-iminodoxorubicin (GPX-150), on human topoisomerase IIβ activity and cardiac function in a chronic rabbit model. Investigational new drugs, 2016. 34(6): p. 693-700.
29.Shalmali, N., M.R. Ali, and S. Bawa, Imidazole: an essential edifice for the identification of new lead compounds and drug development. Mini reviews in medicinal chemistry, 2018. 18(2): p. 142-163.
30.Zhang, M., et al., Design, synthesis and anti-cancer activity of pyrrole-imidazole polyamides through target-downregulation of c-kit gene expression. European Journal of Medicinal Chemistry, 2020. 207: p. 112704.
31.Janiak, C., A critical account on π–π stacking in metal complexes with aromatic nitrogen-containing ligands. Journal of the Chemical Society, Dalton Transactions, 2000(21): p. 3885-3896.
32.Administration, U.S.F.a.D., Novel Drug Approvals for 2020, FDA, Editor. 2020.
33.Pochorovski, I., et al., Quinone‐Based, Redox‐Active Resorcin [4] arene Cavitands. Angewandte Chemie International Edition, 2012. 51(1): p. 262-266.
34.Burmudžija, A., et al., Ferrocenyl based pyrazoline derivatives with vanillic core: synthesis and investigation of their biological properties. RSC advances, 2016. 6(94): p. 91420-91430.
35.Hou, L.-J., et al., YM155 enhances docetaxel efficacy in ovarian cancer. American journal of translational research, 2018. 10(3): p. 696.
36.Gilandoust, M., et al., Synthesis, characterization and cytotoxicity studies of 1, 2, 3-triazoles and 1, 2, 4-triazolo [1, 5-a] pyrimidines in human breast cancer cells. Bioorganic & Medicinal Chemistry Letters, 2018. 28(13): p. 2314-2319.