高良姜根茎抗肿瘤活性成分的筛选、结构修饰及其构效关系研究

Alpinia officinarum Hance, belonging to the genus Alpinia (Zingiberaceae),has been used as a traditional medicine in China (named Galanga) for treating colds , relieving stomach ache and reducing swelling.Modern research showed that this plant has broad antitumor activity and has been recommended as one of natural cancer prevetion herbs.We previous research founded that the flavonoids especically galangin which has the most hingly content in the plant has signigicant anticancer activity through inhibiting part function of Cyclin-Dependent Kinases. There are reports about it's activities,while ,there is little research on the structural modification and Structure–activity relationship of it.Therefore, in this project, we planned to screen the active constituents from the Alpinia officinarum in further , use Mannich-reaction introducting the nitrogen atom D-ring into galangin,evaluate the CDKs inhibition activity and anti-tumor effect of the products and analysis the structure–activity relationship on the basis of our preliminary studies.Through the above studies, we plan to clarify the efficacy material basis and provide scientific evidence for developing highly selectiv and strong affinity CDKs inhibitor as new anticancer drug.

高良姜 (Alpinia officinarum Hance) 为姜科山姜属植物高良姜的干燥根茎，为常用中药，载于历版《中国药典》。具有温胃止呕、散寒止痛的功效。现代研究表明高良姜具有广泛地抗肿瘤活性，被推荐为天然防癌草药之一。本项目组前期研究发现高良姜根茎中黄酮类化合物尤其是含量最高的高良姜素能够抑制部分细胞周期素依赖性蛋白激酶（CDKs）的活性,表现显著的抗肿瘤活性。关于其生物活性研究较多，但关于有效成分结构修饰及其构效关系研究少有报道。本申请项目拟在前期工作的基础上，进一步深入筛选高良姜根茎中的抗肿瘤活性成分，阐明其药效物质基础；以主要成分高良姜素为底物，采用半合成手段引入含氮D环到其母核结构中合成抗癌化合物Flavopiridol 的结构类似物，检测产物CDKs 抑制活性及其抗肿瘤活性，进行构效关系分析，为研发选择性高、亲和力强的CDKs 抑制剂类抗肿瘤药物提供科学依据。

高良姜为姜科 Zingiberaceae 山姜属植物，为我国传统中药。现代药理学研究表明，高良姜具有广泛地抗肿瘤活性，被推荐为天然防癌草药之一。因此，本项目对其抗肿瘤活性成分进行研究。主要研究内容包括：采用MTT法筛选高良姜根茎抗肿瘤活性部位。结果发现，高良姜根茎抗肿瘤活性部位为石油醚萃取部位和二氯甲烷萃取部位；从二氯甲烷萃取部位分离得到一个新的二苯基庚烷类化合物； 合成了6个新的高良姜素含氮衍生物，确定了结构修饰产物的结构。采用MTT法评价结构修饰产物对7种肿瘤细胞的抗癌活性，结果表明化合物2-phenyl-3,5,7-trihydroxy-8-(morpholinomethyl)-4H–chromen- 4-one和2-phenyl-3,5,7-trihydroxy-8-(piperazin-1-ylmethyl)- 4H- chromen-4-one对被检测的细胞株抗肿瘤活性显著优于高良姜素，抗肿瘤活性的增强与8-位含氮杂环的引入有关。本项目再次评价了高良姜素对人乳腺癌细胞MCF-7细胞的增殖抑制作用，首次报道高良姜素诱导MCF-7细胞凋亡。并探讨了其诱导凋亡的作用机制：I. 通过线粒体途径，下调抗凋亡蛋白Bcl-2表达，上调促凋亡蛋白Bax, Bad的表达；显著提高 cleaved-Bid, cleaved-caspase-8, cleaved-PARP, cleaved-caspase-3和cleaved-caspase-9 的表达。II. 抑制PI3K/Akt 信号通路，高良姜素能够显著抑制PI3K和Akt 蛋白的磷酸化。III. 影响细胞周期进程。高良姜素能够下调cyclin D3, cyclin B1, CDK 1, CDK 2, CDK 4 的表达，显著提高p21,p27和p53蛋白的表达。本项目的研究结果丰富了化合物库，为开发新颖的抗肿瘤药物提供物质基础。此外，本项目研究发现高良姜素发挥抑制乳腺癌增殖作用是通过多靶点而不是单靶点，是一种有潜力的可用于防治乳腺癌的化学药物。