基于体内吸收代谢和谱效关系探索小叶莲抗乳腺癌药效物质

The therapeutic effect and quality of traditional Chinese medicine (TCM) are known to directly depend on its effective substances. It is the basic way to reveal the principle of TCM to treat diseases and clarify the effective substance of traditional Chinese medicine by exploring the interaction between the chemical substances and the organism's life activities. In previous studies, we have found that the extract of the Sinopodophylli Fructus has a strong anti breast cancer effect, and its ethyl acetate soluble fraction is the effective part of anti breast cancer. In this program, we will do research in a multi-disciplinary manner by using the methods of pharmacology, TCM analysis, chemistry, and pharmacokinetics. Firstly, we will apply Sinopodophylli Fructus to treat the animals with breast-cancer, and study its in vivo process in the animals simultaneously by LC-MS. Secondly, the potential effective substances of Sinopodophylli Fructus will be proposed by analyzing the relationship between the fingerprints of biosamples and the efficacies of Sinopodophylli Fructus, combined with on-line activity detection. Thirdly, the potential effective substances will be isolated and prepared from Xiaoyelian or biosamples guided by LC-MS. Finally, the pharmacological effects of potential effective substances will be verified by in vitro and in vivo models. This program will not only clarify the in vivo process of Sinopodophylli Fructus in animals with breast cancer, but also clarify the effective substances of anti breast cancer. Furthermore, this program will provide ideas for the research on effective substances of TCM, and to the quality control and clinical application of traditional Chinese medicine has important guiding significance.

中药药效物质与临床疗效和中药质量密切相关，探索中药的化学物质与机体生命活动的交互规律，是揭示中药治疗疾病的原理、阐明中药药效物质的基本途径。前期研究发现小叶莲提取物具有较强的抗乳腺癌作用，其乙酸乙酯萃取部分为抗乳腺癌有效部位。为了阐明小叶莲抗乳腺癌药效物质，本研究拟运用中药药理学、中药分析、中药化学、中药药代动力学多学科交叉的方法，以移植性和药物诱发性乳腺癌动物为模型进行药效学研究，同时采用液质联用技术研究小叶莲原形成分及代谢产物的体内动态分布特征，通过对生物样品指纹谱和效应之间的关系进行分析，结合在线活性检测，推测潜在药效成分，然后在LC-MS的引导下，对潜在的药效成分进行分离制备，并通过体内、体外实验进行活性验证。最终阐明小叶莲有效部位在乳腺癌模型动物体内的药效学、药动学特征和抗乳腺癌的药效物质。本项目将为中药药效物质研究提供思路，对指导中药的质量控制和临床应用具有重要意义。

对小叶莲抗乳腺癌有效部位（Xe）化学成分进行了HPLC-MS分析，指认了102个化合物，主要为木脂素和黄酮类，其中47个为小叶莲首次报道，16个为新化合物，为进一步明确小叶莲抗乳腺癌药效物质奠定了基础。. 正常大鼠和乳腺癌模型大鼠分别灌胃小叶莲的抗乳腺癌有效部位后，分别从其体内指认了原形成分和代谢产物总计共59个和55个，经数据库检索33个代谢产物为新化合物。对比小叶莲抗乳腺癌有效部位在正常大鼠体内和乳腺癌模型大鼠体内的代谢产物分布及代谢类型后发现：小叶莲抗乳腺癌有效部位的原形成分和代谢产物在正常大鼠体内排出体外的数量相对多于乳腺癌模型大鼠，而原形成分和代谢产物的分布在正常大鼠体内的数量相对少于乳腺癌模型大鼠。研究结果表明小叶莲抗乳腺癌有效部位以原形成分和代谢产物的形式在乳腺癌模型大鼠体内发挥治疗作用。. 利用PharmMapper化合物活性预测工具，对首次在小叶莲中报道、且结构确定的23个化合物（8个新化合物）进行活性预测，发现有19个化合物（7个为新化合物）具有潜在的抗癌活性；对7个代谢产物进行活性预测，发现6个代谢产物具有抗癌活性。. 对小叶莲的抗乳腺癌有效部位以及9个主要成分进行了大鼠体内药物动力学研究，通过比较正常大鼠和模型大鼠的药动学参数，初步确定小叶莲抗乳腺癌主要有效成分在乳腺癌模型大鼠体内的药物达峰时间提前，半衰期延后，在较长时间内能维持一定的血药浓度，这些参数说明药物在乳腺癌模型大鼠中能更好的发挥治疗作用。. 通过化学药物DMBA诱导大鼠乳腺癌模型进行小叶莲抗乳腺癌有效部位（Xe）的谱效关系的研究，发现小叶莲的抗乳腺癌有效部位高剂量组（Xe-HD）表现出一定的肿瘤抑制作用。推测山柰酚和槲皮素及其衍生物以及它们的代谢产物为小叶莲抗乳腺癌有效成分。. 根据活性预测结果，结合小叶莲的抗乳腺癌有效部位在大鼠体内的代谢、分布和药物动力学参数，在LC-MS的引导下分离制备了9个潜在药效成分，并发现3个活性强的抗乳腺癌活性成分。