靛玉紅衍生物之合成研究及其抗癌作用
Synthesis of indirubin analogues
as antitumor agents
中文摘要indirubin 藥理作用具有抑制 cyclin-dependent kinase(CDKs)、glycogen synthesis kinase(GSK-3)也是 aryl hydrocarbon receptor (AhR)致效劑。因 indirubin 抑制 CDK 便會造成細胞週期停止在 G1 and G2/M phase，進而抑制細胞增生或是細胞毒 殺。Indirubin 曾在慢性骨髓白血病(CML)臨床試驗發現副作用很少，不會影響造 血功能、肝及腎功能。為改善 indirubin 溶解度不佳，並增加活性，經化學構造 修飾並探討 indirubin 之化學構造與藥效關係。本論文擬合成 indirubin 衍生物並 探討結構與藥效的關係，提供藥物設計者的重要參考。
合成 indirubins 最簡便的方法就是在無氧的情況下結合 indoxyl acetate 和
isatins(63~73)便得到化合物 3、30~39 (產率：70~90%)。利用 3,4 位不同取代的 aniline 加入 chloral hydrate 和 NH2OH HCl 反應得到 isonitrosoacetanilides
(57~62)，以濃硫酸或是 BF3 當催化劑加熱會產生 isatins (63~68)。Indirubins(3、 30~39)用 pyridine 溶解再加入 NH2OH HCl 即能得到 indirubins -3’-oximes(20、 40~49)，產率為 70~80%。 藉由 MTT assay 可發現對於 MCF-7 抗乳癌活性最好的是化合物 49 和 48，IC50 為 9.67、16.9μM；對於 U937 抗白血病活性最好的是化合物 44 和 42 ，IC50 為 3.95、4.67μM，化合物 40、41、43、48、49 仍比化合物 20 有效，化合物 45、 46、47 對兩種細胞均無抑制作用。 英文摘要
Indirubins were found to inhibit the cell cycle regulating cyclin-dependent
kinases(CDKs),glycogen synthesis kinase(GSK-3) and aryl hydrocarbon receptor (AhR) agonist. Indirubins inhibit the proliferation of cells through arresting the cells in the G1 and G2/M phase of the cell cycle by inhibition of CDKs. Indirubins
exhibited good antitumor activity and minor toxicity in chronic myelocytic leukemia clinical trials. Indirubin didn’t affect the production of haematopoietic stem cells nor the function of liver and kidney. To improve the indirubin’s defect of poor solubility and absorption, we design a series of indirubin analogues by chemical modification for its structure. In this paper, we synthesis several indirubin analogues and provide the SAR study for further drug designer.
For synthesis of indiruins(3、30~39), the most convenient method is that dimerization of indoxyl acetate and isatins(63~73).The desired isatins were prepared through using the corresponding 3,4-mono or bisubstituted anilines as starting materials. The
appropriate anilines were reacted with chloral hydrate and hydroxylamine
hydrochloride to give the corresponding isonitrosoacetanilides (57~62), and were heated with c-H2SO4 or BF3 to give desired isatins(63~68)。The indirubins - 3’ – oximes (20、40~49) were synthesized from the indirubins(3、30~39) with
hydroxylamine hydrochloride in pyridine under reflux.(yield：70~80%)
For anti-proliferation of MCF-7(human breast cancer epithelial cell line), the most potent compound of 49 and 48 showed a IC50 of 9.67 and 16.9μM ；For
anti-proliferation of U937(human monocytic leukemia cells), the most potent compound of 44 and 42 showed a IC50 of 3.95 and 4.67μM. Compound 40、41、 43、48、49 were still more potent than compound 20 but compound 45、46、47 were inactive in both MCF-7 and U937 cells.
