靛玉紅衍生物之合成研究及其抗癌作用

中文摘要 indirubin藥理作用具有抑制 cyclin-dependent kinase(CDKs)、 glycogen synthes

is 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。

Synthesis of indirubin analogues as antitumor agents

英文摘要 Indirubins were found to inhibit the cell cycle regulating cyclin-dependent kinases(CDKs),glycogen synthe

sis 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.