金粟兰属植物具抗乳腺癌转移活性的新颖倍半萜成分的研究

The plants of genus Chloranthus are widely distributed in the southern part of China. The most species in Chloranthus are widely used in folk medicine as antispasmodic，Antiseptic and anticancer drugs. Previous phytochemical studies of Chloranthus henryi Hemsl have demonstrated the presence of sesquiterpenoids as well as diterpenoids, flavonoids and coumarins. New sesquiterpenoids derivatives isolated from this plant, which are classified as lindenan type with with a novel big cyclic framework. In addition, these sesquiterpenoids inhibited adhersion of MDA-MB-231 with basement membrane component(Matrigel) significantly, and supressed MDA-MB-231 cells to invade through reconstituted basement membrane markedly. Furthmore, it was found that MDA-MB-231 cells decreased expression of Runx2, a critical metastatic regulator, in dose-dependant manner while treated with these components. It suggest the lindenan type sesquiterpenoids from Chloranthus henryi may be a potent anti-metastatic agent. But the structures of these sesquiterpenoids are complex and it is difficult to isolate these components from the plant of the genus Chloranthus by normal methods. In this project, we isolate the lindenan type sesquiterpenoids from the four species in Chloranthus by the means of molecular imprinted technique and elucidate their structures by spectroscopic methods including 1H-NMR、13C-NMR、1H-1HCOSY、HSQC、HMBC and HRMS experiments. Combining with the anti-metastatic test in vitro and In vivo, The relation between chemical structure and anti-metastatic activity of these sesquiterpenoids will been systematically studied to find anti-metastatic leading compounds. The result of the research will been meaningful to better utilization of these plants and also helpful for the development of a new medicine for breast cancer.

乳腺癌是女性中常见的恶性肿瘤，其癌细胞的转移是乳腺癌复发和死亡的主要原因，从中草药中寻找抗转移的有效成分有着重要意义。金粟兰属的多种植物具有祛风散寒，舒筋强骨，活血散瘀，消肿止痛之功效。民间应用抗癌历史悠久。我们在前期工作中，首次从宽叶金粟兰中分离鉴定了一类结构新颖的乌药烷型倍半萜类化合物，并发现其有较强的抗转移活性，提示该类化合物可能是抗转移有效成分。但该类化合物结构复杂，骨架新颖，常规方法分离和鉴定难度大，现已发现的化合物数量少，难以对该类化合物进行系统的比较和评价。因此，我们将应用分子印迹技术，对金粟兰属4种植物的乌药烷型倍半萜类化合物进行定向分离，以快速、选择性地获得更多该类化合物，配合体内外药理模型筛选其中的有效成分，从分子水平阐明金粟兰属植物抗转移作用的物质基础，通过构效关系和结构修饰研究，寻找新的高效低毒的先导化合物，为开发具有自主知识产权的治疗乳腺癌转移的创新药物奠定基础。

本项目运用现代科学技术与方法，以抗乳腺癌转移活性为导向，对金粟兰属植物宽叶金粟兰（Chloranthus henryi）、多穗金粟兰（C. multistachys）、丝穗金粟兰（C. fortunei）和及己（C. serratus）4种植物中的的倍半萜类化学成分及其活性进行了系统的研究。应用分子印迹及其他现代色谱技术从上述4种金粟兰植物中分离了105个化合物，其中倍半萜类化合物67个，二萜化合物9个，单萜和三萜化合物3个，其他化合物26个，并采用波谱和化学方法鉴定这些化合物的结构。在分离并鉴定的化合物中，宽叶金粟兰内酯A~F、宽叶金粟兰醇E、及己醇A~E、丝穗金粟兰内酯A、丝穗金粟兰酚苷、多穗金粟兰甲素等15个化合物为新化合物，其他大多为首次从该属植物中分离得到。对分离得到的主要化学成分经抗乳腺癌转移活性筛选，有23个倍半萜类化合物显示对乳腺癌细胞MDA-MB-231的抗转移活性，可显著抑制乳腺癌细胞MDA-MB-231的黏附能力和侵袭能力，其分子作用机理可能与其降低乳腺癌细胞内TGF-βR I活性有关。应用分子对接方法模拟有效分子作用机理，并进行定量构效关系分析（3D QSAR）和药效团分析，优选出4个抗乳腺癌转移活性的先导化合物。研究结果进一步确定金粟兰植物中倍半萜类化合物是抗乳腺癌转移活性的主要有效成分，从分子水平阐明了这些中草药抗乳腺癌转移的物质基础，为金粟兰属的资源开发和合理应用提供系统的科学资料，为抗肿瘤新药的研制奠定了基础。