忍冬木层孔菌抗乳腺癌成分分析及作用机制研究

Phellinus lonicerinus is a specy of medicinal Phellinus linteus collected in Quanlity Standard of Chinese Herbal Medicines in Hubei Province, which was found to possess good anti-breast cancer and estrogenic activities. As the estrogenic receptors were the important targets for the treatment of breast cancer, natural selective anti-cancer active estrogen receptor modulators were screened from Chinese medicine through reverse thinking in the project. The natural excellent anti-breast cancer selective estrogenic receptor modulator or compositions were expected to obtain with the aim for solving the similar menopausal syndromes and other adverse effects in the endocrine therapy for breast cancer, and further study the anti-breast cancer and to provide the theoretical basis for the treatment of breast cancer and drug development. The chemical constituents were isolated through column chromatography and preparative HPLC methods. The anti-breast cancer active compounds were screened by MTT methods, and their estrogenic receptor effects were evaluated via cancer cell proliferation assays. Their modulating mechanism were studied in two ways: the binding sides and binding capacities of different compounds with estrogen receptors were calculated by computer-aided docking simulations, the binding capacities were determined through the yeast two-hybrid methods, and the binding sites were also measured by nuclear magnetic resonance technology. The in vitro and in vivo Signaling pathways modulating effects were determined.

忍冬木层孔菌是药用桑黄的品种之一，载于《湖北省中药材质量标准》。本研究发现其具有良好的抗乳腺癌作用，同时有良好的雌激素样作用。雌激素受体是乳腺癌治疗的重要靶标，本课题利用中药优势，采用反向思维方式，从具有抗癌作用的成分中筛选天然选择性雌激素受体调节剂。以期获得优秀的抗乳腺癌天然选择性雌激素受体调节剂或组合物。解决目前乳腺癌内分泌治疗易产生类围绝经期综合症等不良反应的重要基础问题，进一步探索其抗乳腺癌的作用机制，为乳腺癌治疗和新药研究提供理论基础。 采用柱层析分离、高效液相制备等方法分离化合物；MTT法等测定其抗癌作用，细胞增殖实验判断其雌激素调节作用，获得目标化合物；进一步探讨目标化合物雌激素调节作用机制：通过分子对接法计算化合物与雌激素受体的结合能力，采用酵母双杂交等方法测定化合物与雌激素受体的结合力。通过核磁共振技术测定化合物与雌激素受体的结合位点。测定体内外雌激素信号通路调节作用。

对市场常见黄桑品种进行了调查，测定了原儿茶酸、hispolon两种活性成分含量。为控制桑黄质量提供了依据。制定了水马桑药材质量标准，修订了忍冬木层孔菌药材质量标准。收载于《湖北省中药材质量标准》。. 从忍冬木层孔菌及寄主水马桑有效部位中分离、鉴定化合物22个。通过抗癌、雌激素样作用评价及相关性分析。发现hispolon、甲基hispolon抗癌作用良好，雌激素调节作用理想，具有先导化合物特性。. 体内外实验进一步证实甲基hispolon、hispolon，具有雌激素受体部分激动剂特点。雌激素缺乏时，具有雌激素作用，雌激素浓度过高时，具有抗雌激素作用。其作用主要是通过与雌激素受体结合而发挥。高剂量用于治疗妇科肿瘤可克服内分泌治疗易产生类围绝经期综合症等不良反应，低剂量用于治疗围绝经期综合症，可降低诱发乳腺癌等肿瘤的风险。为围绝经期综合症治疗提供了新的思路和方法。