肺癌為最常見的癌症之一，其中 85% 為非小細胞肺癌 (NSCLC)。研究顯示，在約 50% NSCLC 患者中，表皮生長因子受體 (EGFR) 的基因突變 (mutation) (L858R 或 del 19) 造成癌細胞依賴 EGFR 之信號傳遞。EGFR 酪胺酸激酶抑制劑（Iressa® 和 Tarceva®）為治療非小細胞肺癌提供了有效的策略，然而繼發性突變 EGFR T790M 已成為臨床上產生抗藥性的主要原因。因此，我們發展選擇性靶向 EGFR L858R/T790M 的第三代 EGFR 酪胺酸激酶抑制劑，期望將來對有抗藥性的病患能有更好的治療效果。
將化合物 19 的哌啶環進行擴張，並探討嘧啶上取代基的結構與活性關係。另外，我們開發可信任的兩步驟平行合成平台，篩選出對於 EGFR L858R/T790M 有選擇性的一級胺取代基，經由混成設計 (hybrid design)，我們得到了具有高度選擇性的化合物 65，其 EGFR DM 抑制活性提升了 5 倍，而 H1975 細胞活性則是提升了 19 倍。
另一方面，我們將先導化合物 25 進行最佳化，透過環的改變將呋喃[2,3-d]嘧啶換成吡咯[2,3-d]嘧啶，得到化合物 26，其 H1975 細胞活性提升了 3 倍，且選擇性高達 50 倍。然而，化合物 26 溶解度較差，我們將其做成甲基磺酸鹽增加水溶性。

Lung cancer is the most common human cancer, of which 85% presents non-small cell lung cancer (NSCLC). In approximate 50% of NSCLC patients, mutation (L858R or del 19) of epidermal growth factor receptor (EGFR) causes cancer cell dependence on its signaling pathway. The first generation of EGFR tyrosine kinase inhibitors (Iressa® and Tarceva®) provided an effective strategy for the treatment of NSCLC. However, the acquired EGFR T790M mutation has become a leading cause of clinically acquired resistance to these agents. Therefore, we developed irreversible third generation EGFR kinase tyrosine inhibitors selectively targeting EGFR L858R/T790M, which expect to overcome the resistance issues for NSCLC therapy.
The piperidine ring of compound 19 was initially expanded. Then we investigated the structure and activity relationship of the substituents on the pyrimidines. In addition, we established a trusted two-step parallel synthesis platform to screen for primary amine substituents selective for EGFR L858R/T790M. Through hybrid design, we obtained highly selective compound 65 in EGFR DM. The EGFR DM inhibition activity was increased by 5 folds, and the corresponding antiproliferative activity against H1975 cells was improved by 19 times.
On the other hand, we optimized the lead compound 25 through ring variation. We converted the core structure of compound 26 from furano[2,3-d]pyrimidine to pyrrole[2,3-d]pyrimidine and then obtained compound 26. The antiproliferative activity against H1975 cells of compound 26 was increased by 3 folds. Furthermore, it showed 50-fold selectivity over A431 cells. Due to the poor solubility of compound 26 was poor, its mesylate salt was synthesized to improve water solubility.

中文摘要 i
Abstract iii
謝誌 v
目錄 vi
表目錄 x
圖目錄 xi
流程目錄 xiv
縮寫對照表 xv
壹、緒論 1
1.1前言 1
1.2 蛋白質激酶 (Protein kinase) 5
1.3表皮生長因子受體 (Epidermal growth factor receptor, EGFR) 7
1.3.1 EGFR 與肺癌 8
1.3.2 EGFR 結構 11
1.3.3 EGFR 訊息傳遞路徑 12
1.4 EGFR 酪胺酸激酶抑制劑 14
1.4.1 第一代 EGFR 酪胺酸激酶抑制劑 16
1.4.2 第二代 EGFR 酪胺酸激酶抑制劑 19
1.4.3 第三代 EGFR 酪胺酸激酶抑制劑 20
1.4.4 第四代 EGFR 酪胺酸激酶抑制劑 23
貳、研究動機 25
2.1 三環噻吩嘧啶為核心結構之第三代 EGFR TKI初選化合物 篩選 26
2.1.1 BPR1Q0096S0 之開發歷程 26
2.1.2 高通量平行合成平台 27
2.1.3 研究構想 I 29
2.2 呋喃嘧啶為核心結構之第三代 EGFR TKI 先導化合物最佳化 30
2.2.1 BPR3K0360S0之開發歷程 30
2.2.2 藥物動力學的探討 32
2.2.3 研究構想 II 34
2.3 生物活性測試分析 35
2.3.1 酵素抑制活性測試 36
2.3.2 癌細胞株生長抑制活性測試 37
參、結果與討論 39
3.1 三環噻吩嘧啶為核心結構之第三代 EGFR TKI初選化合物篩選 39
3.1.1 化合物21之合成 39
3.1.2 環大小修飾 (Ring variation) 40
3.1.3 嘧啶上取代基之SAR 43
3.1.4 兩步驟平行合成平台 49
3.1.4.1 核心結構 63 的合成 49
3.1.4.2 平行合成方法 50
3.1.4.3平行合成平台之可信度 52
3.1.5 平行合成結果與活性分析 54
3.1.6 初選化合物 65 59
3.1.7 化合物 65 變溫核磁共振實驗 61
3.1.8 化合物 65 之代謝穩定性 63
3.2 呋喃嘧啶為核心結構之第三代 EGFR TKI 先導化合物最佳化 64
3.2.1 化合物 75 的合成 64
3.2.2 化合物 82 的合成 65
3.2.3 化合物 26 的合成 66
3.2.4 化合物 97 的合成 68
3.2.5 化合物 25 之衍生物的活性分析 68
3.2.6 改善化合物 26 之溶解度 70
肆、總結 72
4.1 三環噻吩嘧啶為核心結構之第三代 EGFR TKI初選化合物 72
4.2 未來展望 I 73
4.3 呋喃嘧啶為核心結構之第三代 EGFR TKI 先導化合物最佳化 74
4.4 未來展望 II 76
伍、實驗部分 77
5.1 一般實驗方法 77
5.2 化合物之實驗步驟與光譜資料 80
陸、參考資料 172

附錄一 化合物之核磁共振光譜 177
附錄二 化合物之編號對照表 244
附錄三 生物活性測試方法 247
附錄四 論文口試投影片 253

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