臺灣博碩士論文加值系統

本實驗室依據先前合成benzothieno[2,3-c]quinolone化合物作為新型抗癌藥，經放療篩選，發現其benzothiophene之中間產物，具有強力增敏及保護放療作用。另外，文獻報導thiophene衍生物具抗炎活性，故本實驗室設計合成一系列benzothiophene及thiophene衍生物，以期開發高效低毒的新型放療增敏保護與抗炎藥。
分別以對甲氧基桂皮酸 (p-methoxycinnamic acid)、苯并噻吩-2-羧酸 (benzo[b]thiophene-2-carboxylic acid)、2-噻吩甲酸 (2-thiophenecarboxylic acid)、2-噻吩-3-烯酸 (3-(2-thienyl)acrylic acid)、2-噻吩乙酸 (2-thiopheneacetic acid)、2,5-噻吩二甲酸 (2,5-thiophenedicarboxylic acid)為起始物，加入過量亞磺醯氯 (thionyl chloride)於甲苯中迴流1-48小時而得到含醯氯之中間產物，未經純化下，即直接加入anilines反應而得到標的產物。
經初步活性試驗，合成之化合物中，皆具良好放療增敏效果，以T3c (48.2 %)最佳。另外，由MTS分析方法之結果顯示，化合物對於MCF-7、NCI-H460、SF-268三種癌細胞株在正常含氧條件之毒性試驗無明顯抗癌效果。而在鹿角菜誘導發炎動物模式進行抗炎藥物篩選中，化合物5c、5d、M3c、T3c抗炎活性較ibuprofen佳。

Our laboratory previously carried out the design and synthesis of a series of benzothieno[2,3-c]quinolone derivatives to develop novel anticancer drugs in our laboratories. After screening of the radiosensitizer/radioprotectant, the results showed that benzothiophene intermediates had strong potency to enhance the activity of radiation. On the other hand, literatures indicated that some thiophene derivatives possess significant antiinflammatory activity. Therefore, we further designed and synthesized a series of benzothiophene and thiophene derivatives to develop novel highly effective and low toxic radiosensitizers/radioprotectants and antiinflammatory drugs.
The starting materials, p-methoxycinnamic acid, benzo[b]thiophene-2-carboxylic acid, 2-thiophenecarboxylic acid, 3-(2-thienyl)acrylic acid, 2-thiopheneacetic acid, and 2,5-thiophenedicarboxylic acid, were respectively treated with excess thionyl chloride in toluene to the preparation of all the target compounds. After heating at reflux for 1-48 hours and being concentrated in vacuum, acyl intermediates without purification were further reacted with various anilines in the presence of triethylamine to afford the target compounds.
The preliminary results showed that benzothiophene and thiophene derivatives revealed potent radiosensization, especially T3c (48.2 %) superior to others. However, the results of target compounds by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl) -2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay showed that none of the synthetic products were active against MCF-7 (breast carcinoma), NCI-H460 (lung cancer), or SF-268 (glioblastoma) cancer cell lines. On the other hand, compounds 5c、5d、M3c、T3c were much potent than ibuprofen after screening of the target compounds by carrageenan-induced paw edema model.

目錄
頁次
正文目錄………………………………………………………………..Ⅰ
附圖目錄………………………………………………………………..Ⅲ
中文摘要…………………………………………………………..……Ⅶ
英文摘要……………………………………………………………….Ⅷ

第一章、緒論………………………………………….………………….1
第一節、前言…………………………………….…………………..1
壹、癌症簡介………………………………..………………….1
貳、癌症治療藥物的方法……………..……………………….2
參、放射線……………………………..……………………….3
肆、放射線治療………………………..……………………….6
第二節、研究目的………………………….………………………..9
壹、放療增敏和保護劑………………..……………………….9
貳、研究目的…………………………….……………………18
第三節、研究方法…………………………….……………………22
壹、藥物設計原理.……………………….…………………...22
貳、合成反應概要……………………….……………………24
第二章、材料與方法……………………………………………………28
第一節、試藥來源…………………………………………………28
第二節、重要儀器…………………………………………………30
第三節、合成實驗…………………………………………………31
第四節、生物活性試驗……………………………………………44
壹、放療增敏/放療保護試驗…...……………………………44
貳、抗腫瘤活性試驗……………………….………………...45
參、抗炎活性試驗…………………………..………………..47
第三章、結果……………………………………………………………49
第一節、化學合成…………………………………………………49
第二節、生物活性…………………………………………………92
壹、放療增敏/放療保護試驗…...……………………………92貳、抗腫瘤活性試驗……………………….………………...94
參、抗炎活性試驗………………………………….………..96
第四章、討論…………………………………………………..………102
第一節、化學合成………………………………………..………102
第二節、生物活性………………………….…………………….124
第五章、結論…………………………………….……………….……129
第六章、參考文獻……………………………….…………………….130














表目錄
頁次
表一、Effect of 3a-3s and 3a＊-3b＊ on the cell viability in Hela
cell……………………………………………………..………..19
表二、Antiinflammatory activity of 1-7….……………………….…..…20
表三、Effect of 4a – F7d on the cell viability in Hela cell…………...…92
表四、Effect of C3a – B8a on the cell viability in Hela cell…………....93
表五、Effect of T3a - D3d on the cell viability in Hela cell……….……93
表六、Effect of 4a – B8a on the cell viability in MCF-7, NCI-H460 and
SF-268…………………………………….………………….…94
表七、Effect of T3a - D3d on the cell viability in MCF-7, NCI-H460 and
SF-268…………………………………….………………….…95
表八、Antiinflammatory activity of 4a - 5d at dose 20 mg/kg i.p. for SD
rats…………………………………….……………...…………97
表九、Antiinflammatory activity of F7a - B8a at dose 20 mg/kg i.p. for
SD rats………………………….……………...…………….…98
表十、Antiinflammatory activity of T3a - M3d at dose 20 mg/kg i.p. for
SD rats………………………….……………...…………….…99
表十一、Antiinflammatory activity of D3a-D3d at dose 20 mg/kg i.p. for
SD rats……………………….………..……...…………….…100
表十二、Antiinflammatory activity of 5d、B8a、C3d、A3a、A3b、
M3c、M3d at dose 20 mg/kg i.p. for SD rats...………………..101
表十三、Antiinflammatory activity of
N-(Substituted-phenyl)-3-carboxamidyl-5-methylisoazoles at
dose 10 mg/kg i.p. for SD rats………………………………...117
表十四、Antiinflammatory activity of T3e - T3i at dose 20 mg/kg i.p. for
SD rats…………………………………………………….…..127
表十五、Antiinflammatory activity of T3e、T3f、T3h at dose 20 mg/kg
i.p. for SD rats………………………………………....….…..128



圖目錄
頁次圖一、癌細胞的形成…………………..…………………………………1
圖二、由鐳元素產生磁場而放出α、β與γ等射線…………………..3
圖三、直接及間接作用……………………….…………..……………...6
圖四、增敏及保護作用………………………….………………...……..9
圖五、鹵素嘧啶放射增敏劑結構………………………………....……11
圖六、缺氧細胞…………………………………………………………12
圖七、Hypoxic cell radiosensitizers的還原作用…………………….....13
圖八、Hypoxic cytotoxic agents的作用……………………..…….……14
圖九、Amifostine的去磷酸化作用……………………………..………16
圖十、Benzothieno[2,3-c]quinolone衍生物的合成………………….…18
圖十一、放療增敏/放療保護實驗步驟………………………..…….…44
圖十二、MTS與succinate dehydrogenase作用的結構變化圖………..45
圖十三、MTS分析方法實驗步驟…………………………...…………46
圖十四、Carrageenan誘發足蹠浮腫之事前給藥實驗步驟………..….48
圖十五、Structures of T3a and leflunomide analogs……………..……115
圖十六、化合物2之1H-NMR＆13C圖譜………………..……………134
圖十七、化合物4a之1H-NMR＆13C圖譜……………………..…..…135
圖十八、化合物4b之1H-NMR＆13C圖譜…………..…………..……136
圖十九、化合物4c之1H-NMR＆13C圖譜……………..…………...…137
圖二十、化合物4d之1H-NMR＆13C圖譜………………..…………..138
圖二十一、化合物3之1H-NMR＆13C圖譜………………………..…139
圖二十二、化合物5a之1H-NMR＆13C圖譜…………………………140
圖二十三、化合物5b之1H-NMR＆13C圖譜…………………………141
圖二十四、化合物5c之1H-NMR＆13C圖譜………………..……...…142
圖二十五、化合物5d之1H-NMR＆13C圖譜………..…….….………143
圖二十六、化合物F3之1H-NMR＆13C圖譜…………………………144
圖二十七、化合物F4之1H-NMR＆13C圖譜…………….………...…145
圖二十八、化合物F5之1H-NMR＆13C圖譜……………….……...…146
圖二十九、化合物F7a之1H-NMR＆13C圖譜……………………..…147
圖三十、化合物F7b之1H-NMR＆13C圖譜…………………..………148
圖三十一、化合物F7c之1H-NMR＆13C圖譜…………………..……149
圖三十二、化合物F7d之1H-NMR＆13C圖譜……………..…………150
圖三十三、化合物C3a之1H-NMR＆13C圖譜………..….……..……151
圖三十四、化合物C3b之1H-NMR＆13C圖譜………………….……152
圖三十五、化合物C3c之1H-NMR＆13C圖譜…………………..……153
圖三十六、化合物C3d之1H-NMR＆13C圖譜………………….……154
圖三十七、化合物T3a之1H-NMR＆13C圖譜……..…………………155
圖三十八、化合物T3b之1H-NMR＆13C圖譜…………………….…156
圖三十九、化合物T3c之1H-NMR＆13C圖譜……………………..…157
圖四十、化合物T3d之1H-NMR＆13C圖譜………………….………158
圖四十一、化合物M3a之1H-NMR＆13C圖譜………………….……159
圖四十二、化合物M3b之1H-NMR＆13C圖譜………………………160
圖四十三、化合物M3c之1H-NMR＆13C圖譜…………………….…161
圖四十四、化合物M3d之1H-NMR＆13C圖譜………………………162
圖四十五、化合物A3a之1H-NMR＆13C圖譜……………….………163
圖四十六、化合物A3b之1H-NMR＆13C圖譜………………….……164
圖四十七、化合物A3c之1H-NMR＆13C圖譜………………………..165
圖四十八、化合物A3d之1H-NMR＆13C圖譜……………………….166
圖四十九、化合物D3a之1H-NMR＆13C圖譜……………………….167
圖五十、化合物D3b之1H-NMR＆13C圖譜………………………….168
圖五十一、化合物D3c之1H-NMR＆13C圖譜…………………..……169
圖五十二、化合物D3d之1H-NMR＆13C圖譜……………………….170
圖五十三、化合物B6a之1H-NMR＆13C圖譜…………………..……171
圖五十四、化合物B7a之1H-NMR＆13C圖譜………………………..172
圖五十五、化合物B8a之1H-NMR＆13C圖譜………………………..173
圖五十六、化合物B9之1H-NMR＆13C圖譜…………………………174
圖五十七、化合E2之1H-NMR＆13C圖譜……………………………175
圖五十八、化合T3e之1H-NMR＆13C圖譜…………………………..176
圖五十九、化合T3f之1H-NMR＆13C圖譜…………………………..177
圖六十、化合T3g之1H-NMR＆13C圖譜………………………….….178
圖六十一、化合T3h之1H-NMR＆13C圖譜……………………….…179
圖六十二、化合T3i之1H-NMR＆13C圖譜…………………………...180

1.http://www.doh.gov.tw/statistic/data/死因摘要/93年/統計圖/2.主要死因死亡率圖.xls
2.健康教育叢書。第三輯。
3.http://library.med.nyu.edu/patient/hicup/documents/Cancer_Care.pdf.
4.http://www.doh.gov.tw/lane/health_edu/i1.html
5.American Cancer Society®, A Guide to Living with Cancer. 1-40.
6.http://www.csh.org.tw/into/rtweb/information/1.htm
7.李文權。科學知識。第56卷。頁數：79-86。
8.許文林。放射治療的基本原理。國防醫學。第七卷。第四期。1988。頁數:31-334。
9.Tewari, K. S.; DiSaia, P. J. Radiation therapy for gynecologic cancer. J Obstet Gynaecol Res, 2002, 28, 123–140.
10.Alsheyab, F. Implications of cellular and molecular advances on radiation oncology: A Review. Int J Sci Res, 2005, 15, 1-6.
11.Coleman, C. N.; Turrisi, A. T. Radiation and chemotherapy sensitizers and protectors. Oncol Hematol, 1990, 10, 3, 225-252.
12.Walkerd, M. A review of drugs that may be used in conjunction with radiotherapy. Vet Radio, 1982, 23, 5, 220-222.
13.許文林。放射線治療合併化學治療的原則。國防醫學。第十三卷。第六期。 1991。頁數:526—531。
14.Poggi, M. M.; Coleman, C. N.; Mitchell, J. B. Sensitizers and protectors of radiation and chemotherapy. Curr Probl Cancer, 2001, 25, 331-411.
15.McGinn, C. J.; Shewach, D. S.; Lawrence, T. S. Radiosensitizing nucleosides. J Natl Cancer Inst, 1996, 88, 17, 4, 1193-1203.
16.Cecchini, S.; Girouard, S.; Huels, M. A.; Sanche, L.; Hunting, D. J. Single-strand-specific radiosensitization of DNA by bromodeoxyuridine. Radiat Res, 2004, 162, 604–615.
17.Wang, Y.; Pantelias, G. E.; Iliakis, G.. Mechanism of radiosensitization by halogenated pyrimidines: the contribution of excess DNA and chromosome damage in BrdU radiosensitization by minimal in plateau-phase cells. Int J Radiat Biol, 1994, 66, 2,133-142.
18.Jones, G. D.D.; Ward, J. F.; Limoli, C. L.; Moyer, D. J; Aguilera, J. A. Mechanisms of radiosensitization in iododeoxyuridine-substituted cells. Int J Radiat Biol, 1995, 67, 6, 647-653.
19.Hori, H.; Nagasawa, H.; Uto, Y.; Ohkura, K.; Kirk, K. L.; Uehara, Y.; Shimamura, M. Design of hypoxia-targeting protein tyrosine kinase inhibitor using an innovative pharmacophore 2-methylene-4-cyclopentene-1,3-dione. Biochim Biophys Acta, 2004, 1697, 29–38.
20.Airley, R. E.; Monaghan, J. E.; Stratford, I. J. Hypoxia and disease: opportunities for novel diagnostic and therapeutic prodrug strategies. Pharmacogenomics J, 2000, 264, 7094, 666-673.
21.Denny, W. A. The role of hypoxia-activated prodrug in cancer therapy. Lancet Oncol, 2000, 1, 9, 25-29.
22.de Abreu, F. C.; Ferraza, P. A. de L.; Goulart, M. O. F. Some Applications of electrochemistry in biomedical chemistry. Emphasis on the correlation of electrochemical and bioactive Properties. J Braz Chem Soc, 2002, 13, 1, 19-35.
23.Kovacic, P.; Osuna, J. A. Mechanisms of anti-cancer agents: Emphasis on oxidative stress and electron transfer. Curr Pharm Des, 2000, 6, 277-309.
24.Brown, J. M. The hypoxic cell: A target for selective cancer therapy. Cancer Res, 1999, 59, 5863–5870.
25.Denny, W. A. The role of hypoxia-activated prodrug in cancer therapy. Lancet Oncol 2000, 1, 9, 25-29.
26.Kizaka-Kondoh, S.; Inoue, M.; Harada, H.; Hiraoka, M. Tumor hypoxia: A target for selective cancer therapy. Cancer Sci, 2003, 94, 12, 1021-1028.
27.Abbasakoor, F.; Vaizey, C. J.; Boulos, P. B. Improving the morbidity of anorectal injury from pelvic radiotherapy. Colorectal Dis, 2005, 8, 2-10.
28.Varanda, E. A.; Tavares, D. C. Radioprotection: mechanisms and radioprotective agents including honeybee venom. J Venom Anim Toxins, 4, 1, 1998, 5-21.
29.王令瑋。放射線及化學治癌藥物保護劑。臨床醫學。第四十七卷。第二期。頁數:92-95。
30.Spencer CM; Goa KL. Amifostine: a review of its pharmacodynamic and pharmacokinetic properties and therapeutic potential as a radioprotector and cytotoxic chemoprotector. Drugs, 1995, 50, 1001-1031.
31.Budd, G. T.; Garapathy, R.; Adelstein, D. J. Randomized trial of carboplatin plus amifostine versus carboplatin alone in patients with advanced solid tumors. Cancer, 1997, 80, 1134-1140.
32.郭儷萱、朱麗鈴。細胞保護劑Amifostine-癌症患者的福音。彰基藥訊。第29期。第九卷。二期。頁數:1-13。
33.顏慧如。第二部分Benzothieno[2,3-c]quinolone衍生物之合成與抗癌活性評估。碩士論文。2005。頁數:1-76。
34.Svoboda, J.; Stadler, M.; Jandera, A.; Panajotova, V.; Kuchar, M. Synthesis and biological evaluation of new antiinflammatory 1-benzthiophene-2-carboxanilides. Collect Czech Chem Commun, 2000, 65, 1082-1092.
35.CellTiter 96® Aqueous one solution cell proliferation assay.
36.Chao, J. CJ; Chu, C. C. Effects of ginkgo biloba extract on cell proliferation and cytotoxicity in human hepatocellular carcinoma cells. World J Gastroenterol, 2004, 10, 1, 37-41.
37.O’Toole, S. A.; Sheppard, B. L.; McGuinness, E. P.J.; Gleeson, N. C.; Yoneda, M.; Bonnar, J. The MTS assay as an indicator of chemosensitivity/resistance in malignant gynaecological tumours. Cancer Detect Prev, 2003, 27, 47–54.
38.Winter, C. A.; Risley, E.A.; Nuss, G. W. Carrageenin-induced edema in hind paw of the rat as an assay for anti-inflammatory drugs. Proc Soc Exp Biol Med, 1962, 111, 544-547.
39.Di Rosa M.; Sorrentino L. The mechanism of the inflammatory effect of carrageenin. Eur J Pharmacol, 1968, 4, 3, 340-342.
40.Vinegar, R.; Schreiber, W.; Hugo, R. Biphasic development of carrageenin edema in rats. J Pharmacol Exp Ther, 1969, 166, 1, 96-103.
41.Rosa, M. D. Biological properties of carrageenan. J Pharm Pharmacol, 1972, 24, 89-102.
42.Gentilli, M.; Mazoit, J. X.; Bouaziz, H.; Fletcher, D.; Casper, R.F.; Benhamou, D.; Savouret, J. F. Resveratrol decreases hyperalgesia induced by carrageenan in the rat hind paw. Life Sci, 2001, 2, 68, 11, 1317-21.
43.Wright, Jr. W. B.; Brabander, H. J. The Preparation of 3-chlorobenzo[b]thiophene derivatives from cinnamic acids. J Heterocyclic Chem, 1971, 8, 711-714.
44.Castle, S. L.; Buckhaults, P. J.; Baldwin, L. J.; Mckenney, Jr., J. D.; Castle, R. N. The Synthesis of monomethoxy[1]benzothieno[2,3-c]quinolines. J Heterocyclic Chem, 1987, 24, 1103-1108.
45.Lee, C. K.; Yu, J. S.; Ji, Y. R. Determintion of aromaticity indices of thiophene and furan by nuclear magnetic resonance spectroscopic analysis of their anilides. J Heterocyclic Chem, 2002, 39, 1219-1228.
46.Sura, T. P.; MacDowell, Denis, W. H. Cope rearrangements in the benzo[b]thiophene series. J Org Chem, 1993, 58, 4360-4369.
47.Ried, W.; Bender, H. Notiz über einige neue abkömmlinge des thionaphthens. Chem Ber, 1955, 88, 34, 34-38.
48.Jérémie Fournier Dit Chabert; Lionel Joucla; Emilie David; Marc Lemaire An efficient phosphine-free palladium coupling for the synthesis of new 2-arylbenzo[b]thiophenes. Tetrahedron, 2004, 60, 3221-3230.
49.Sall, D. J.; Bailey, D. L.; Bastian, J. A.; etal. Diamino benzo[b]thiophene derivatives as a novel class of active site directed thrombin inhibitors. 5. Potency, Efficacy, and Pharmacokinetic Properties of Modified C-3 Side Chain Derivatives. J Med Chem, 2000, 43, 649-663.
50.AboulWafa, O. M.; Berto, F. A. G. Benzo[b]thiophenes, Part Ⅰ: Synthesis and antimicrobial activity of benzo[b]thienyl-1,3,4-triazoline, and-thiazoline derivatives. Arch Pharm, 1992, 325, 123-127.
51.Reinecke, M. G.; Newsom, J. G.; Almqvist, K. A. An Improved synthesis of thiophene-2,3-dicarboxylic acid by sequential carboxylation. Synthesis, 1980, 4, 327-329.
52.John, J. A.; Tour, J. M. Synthesis of polyphenylene derivatives by thermolysis of enediynes and dialkynylaromatic monomers. Tetrahedron, 1997, 53, 45, 15515-15534.
53.楊巧麗。Leflunomide類似物之合成與抗炎活性。碩士論文。2000。頁數:1-114。