開發新穎EGFR激酶抑制劑於抗癌藥物之研究

Discovery of Novel EGFR Kinase Inhibitors as Anticancer Agents

吳佳憲(Chia-Hsien Wu)

指導教授 ： 廖俊臣 ； 謝興邦

國立清華大學/理學院/化學系/博士(2010年)

摘要

肺癌是當前對人類的健康和生命最具威脅的惡性腫瘤，其中非小 細胞肺癌 (NSCLC)佔了約80% ~ 85%。研究顯示上皮生長因子受器 (EGFR)的過量表現和NSCLC 之間有相當密切的關連。EGFR 為一膜 蛋白，包含了膜外的配體 (ligand)結合受器及膜內的酪胺酸激酶 (tyrosine kinase)。愛瑞莎 (Iressa)和得舒緩 (Tarceva)是兩個通過美國 FDA 的NSCLC 上市藥物，藉由和三磷酸腺苷 (ATP)競爭EGFR 激酶 的活性位置來抑制激酶的活性。但服用愛瑞莎或得舒緩顯現療效的病 患，平均6–12 個月即會產生抗藥性，其中約50%的病患是因為單點 突變 (T790M)造成抗藥性，而使藥物失去效用，因此我們的目標是 發展兼具有一般EGFR 及突變EGFR 激酶抑制活性的化合物，期望在 將來對有抗藥性的病患能提供一個更佳的治療方法。 藉由混成設計(hybrid design)及循理性設計(knowledge-based design)，引入具有立體中心的(S)-phenyl-glycinol 及Michael acceptor 至高通量細胞篩選所獲得出的先導化合物43 中，得到一系列化合物 135，150，175 及178，對EGFR 激酶酵素及EGFR 過度表現的HCC827 肺癌細胞株皆具有1~9 nM 抑制活性，且對愛瑞莎抗藥性的雙突變 EGFR (T790M/L858R)激酶酵素也有10~305 nM 的抑制效果。在結構 與活性的研究中得知，具有立體中心的(S)-phenyl-glycinol 及Michael acceptor 在抑制EGFR 激酶活性上皆是不可或缺的重要因素，將於論 文中詳述，部分結果已發表於J. Med. Chem. 2010, 53, 7316–7326。

關鍵字

上皮生長因子受器 ； 激酶抑制劑 ； 非小細胞肺癌

並列摘要

Lung cancer is the first leading cause of death in human cancer, with 80-85% being non-small cell lung cancer (NSCLC). Over-expression of epidermal growth factor receptors (EGFR) is in 40-80% of NSCLC. Gefitinib (Iressa; AstraZeneca Pharmaceuticals) and erlotinib (Tarceva; Genentech, Inc.) were approved by U.S. Food and Drug Administration as EGFR kinase inhibitors for the treatment of adenocarcinoma (subtype of NSCLC). Gefitinib and erlotinib competitively bind to the ATP binding pocket of EGFR tyrosine kinase domain and inhibit its activity. These two drugs showed high response rates in specific subsets of NSCLC patients, but patients become resistant to treatment with gefitinib or erlotinib after 6-12 months. For approximately 50% of patients who respond initially to gefitinib or erlotinib, drug resistance occurs as a result of secondary mutation such as the Thr790 to Met790 (T790M) mutation. We designed and synthesized novel EGFR-TKIs, and expect that we can overcome the resistance issues for potential NSCLC therapy, and developed 2nd generation drugs.
Through hybrid design and knowledge-based design concepts, (S)-phenyl-glycinol and Michael acceptor groups were introduced to high throughput hit 43 to obtain compounds 135, 150, 175 and 178 with 1~9 nM inhibition against wild type EGFR enzyme and EGFR overexpressed HCC827 NSCLC cell-line. This series also showed 10~305 nM inhibition against gefitinib-resistant double mutant EGFR (T790M/L858R) enzyme. More than 150 analogues based on compound 117 were synthesized, presence of (S)-phenyl-glycinol and Michael acceptor groups are essential for activity in this series of compounds and part of the results were published in J. Med. Chem., 2010, 53, 7316–7326.

並列關鍵字

EGFR ； kinase inhibitor ； NSCLC

參考文獻

83. Lin, W. H.; Song, J. S.; Chang, T. Y.; Chang, C. Y.; Fu, Y. N.; Yeh, C. L.; Wu, S. H.; Huang, Y. W.; Fang, M. Y.; Lien, T. W.; Hsieh, H. P.; Chao, Y. S.; Huang, S. F.; Tsai, S. F.; Wang, L. M.; Hsu, J. T.; Chen, Y. R. A cell-based high-throughput screen for epidermal growth factor receptor pathway inhibitors. Anal. Biochem. 2008, 377, 89-94.

7. (a) Kopka, M. L.; Yoon, C.; Goodsell, D.; Pjura, P.; Dickerson, R. E. The molecular origin of DNA-drug specificity in netropsin and distamycin. Proc. Natl. Acad. Sci. U. S. A. 1985, 82, 1376-1380; (b) Finlay, A. C.; Hochstein, F. A.; Sobin, B. A.; Murphy, F. X. Netropsin, a New Antibiotic Produced by a Streptomyces. Journal of the American Chemical Society 1951, 73, 341-343.

62. Rabindran, S. K.; Discafani, C. M.; Rosfjord, E. C.; Baxter, M.; Floyd, M. B.; Golas, J.; Hallett, W. A.; Johnson, B. D.; Nilakantan, R.; Overbeek, E.; Reich, M. F.; Shen, R.; Shi, X.; Tsou, H. R.; Wang, Y. F.; Wissner, A. Antitumor activity of HKI-272, an orally active, irreversible inhibitor of the HER-2 tyrosine kinase. Cancer Res. 2004, 64, 3958-3965.

1. Hayflick, L.; Moorhead, P. S., The serial cultivation of human diploid cell strains. Exp Cell Res 1961, 25, 585-621.

5. Tan, C.; Tasaka, H.; Yu, K. P.; Murphy, M. L.; Karnofsky, D. A. Daunomycin, an antitumor antibiotic, in the treatment of neoplastic disease. Clinical evaluation with special reference to childhood leukemia. Cancer 1967, 20, 333-353.