基于表面等离子共振原理实时跟踪分子与靶标间相互作用的Biacore系统，是靶向追踪微量海洋活性物质的新方法。蛋白激酶CDK-2是抗肿瘤药物的重要作用靶点。申请人建立了以CDK-2为靶标的Biacore筛选方法，从39种西沙海绵的117个萃取物中筛选出三种海绵具有CDK-2抑制活性。结合UPLC-DAD-QTOF-MS分子特征波谱/色谱在线检测和Biacore活性追踪的方法，已分离得到化合物56个(新化合物38个,1个5/6/4/5环骈合的二萜新骨架),其中CDK-2抑制活性化合物11个。本课题拟分离鉴定这三种海绵中具有CDK-2抑制活性的化合物,从分子、细胞、动物三个层次评价其抗肿瘤活性。采用分子生物学手段,深入研究活性化合物对肿瘤细胞增殖、凋亡、DNA损伤及对CDK-2上下游信号通路的影响,结合计算化学和结构生物学方法,阐明其抗肿瘤作用机制,获得1-2个靶向CDK-2的抗肿瘤先导化合物。

Biacore system that based on the Surface Plasmon Resonance principle is a new way of target-guided tracking for trace bioactive marine natural products. CDK-2 kinases is a critical antitumor drug target. The applicant has established a Biacore screening model targeted on CDK-2. The applicant has found three sponge extracts showed CDK-2 inhibitory activity from 117 extracts of 39 Xisha sponges. By using the methods of UPLC-DAD-QTOF-MS characteristic spectral/chromatography detection and bio-guided tracking based on Biacore, the applicant has obtained 56 compounds (38 new compounds, one new skeleton diterpene with 5/6/4/5 ring system),including 11 compounds with CDK-2 inhibitory activity. This project intend to track and identify the CDK-2 inhibitory compounds from these three Xisha sponges, the isolated compounds will be evaluated their antitumor activities on three levels including cellular, molecular, and animal models. The hit compounds will be studied their effects on tumor cell proliferation, apoptosis, DNA damage and CDK-2 signal pathway by using the cellular and molecular biology methods. On this basis, the applicant will combine computational chemistry with structural biology method to clarify the bioactive compound's antitumor mechanism of action, to obtain 1-2 antitumor leads targeting to CDK-2.