靶向求源策略发掘GRP78受体高亲和中药成分及其分子机理研究

The search for chemotherapy pharmaceuticals from Traditional Chinese Medicine(TCM)，which are targeting, high-efficient, low-toxic and low-cost, has become one of the hottest topic in cancer therapy research nowadays. Having highly specific expression in the cell membranes in tumors and other diseases, GRP78 is a key protein for the onset and the development of diseases and a multi-functional receptor that attracts much concern. Therefore, we successfully established a platform, which combines novel cell membrane affinity chromatography, nanoflow liquid chromatography and mass spectrum technology,in order to search GRP78 ligand from TCM with anti-cancer activity. The GRP78 highly experessed cell membrane were effectively coated on the specially designed phospholipid membrane skeleton, which exhibted good biological compatibility and significant simplicity in preparation and chemical modification,. This novel platform facilitates the high-performance screening of CRP78 affinity compositions from anti-tumor TCM. Coupled with the programmed sample collection of offline two-dimensional liquid chromatography, flow cytometry, gel electrophoresis -tandem mass spectrum and other technologies, this platform can be applied to the efficacy evaluation and molecular mechanism study of the affinity composition cluster. It can aviod the loss of the trace novel-structured compouds and weak affinity composition, and thus benefit the search for some GRP78 targeting candidates.This research can provide not only an effective tool for the discovery of novel targeting tumor-therapy-pharmaceuticals with high efficacy and low toxicity, but also explantions for the mechanism of synergy effects and tenuation of TCM. It finally will provid significant reference for researches aimed at the reduction of drug resistance of tumor and the improvement of living quality of patients.

从中药中寻找靶向、高效、低毒、廉价的化疗或治疗药物（组合物）成为当今肿瘤研究的热点之一。GRP78在肿瘤等疾病的细胞膜表面特异性显著高表达，是其发生发展的关键信号因子，亦是备受关注的多功能受体标靶。为此，我们以GRP78高低表达细胞膜和制备简单、易于化学修饰、生物兼容性好等优点的磷脂膜骨架为材料，成功构建了新型细胞膜亲和色谱-纳流液相色谱-质谱联用技术平台，可对抗肿瘤中药及组方中GRP78高亲和成分及组合物进行高效筛选，结合二维液相程序收集、流式细胞术、凝胶电泳-串联质谱等技术对亲和成分群进行药效评价及分子机理研究。本课题建立的新型细胞膜亲和色谱-纳流液相-质谱联用的技术平台，将减少微量新奇结构化合物和弱亲和组分的丢失，寻优一些靶向GRP78的候选化合物/组合物，为肿瘤治疗提供一些靶向、高效和低毒的新药，有望阐释中药抗肿瘤的部分增效减毒机制，为减少肿瘤耐药性和提高患者生存质量提供重要参考。

寻找高效、低毒、廉价的化疗组合物成为当今肿瘤研究的热点之一。近年来，GRP78 是肿瘤等疾病的细胞膜表面特异性显著高表达，备受关注的多功能受体标靶。寻找GRP78 受体的抑制剂是一个新的抗肿瘤治疗方向。鉴于这些背景，本课题采用高表达GRP78受体的细胞膜为基质，结合仿生双层磷脂膜的亲和特性，成功构建了新型细胞膜亲和色谱筛选平台，可对抗肿瘤中药及组方中GRP78 高亲和成分及组合物进行高效筛选，发现30多个可以结合的中药组分，MS鉴定获得14个化合物，通过MTT活性评价发现，其中6个成分具有显著的抗癌效应，且对顺铂的化疗敏感性明显增强，更难得的是化合物isoliquiritigenin靶向结合GRP78的相互作用和增效抗癌机理研究被香港大学的学者证实。故寻优一些靶向GRP78 的候选化合物/组合物，有望为靶向恶性肿瘤的治疗提供一些高效低毒的新药候选分子，同时为阐释中药抗肿瘤的部分增效减毒机制，为减少肿瘤耐药性和提高患者生存质量提供重要科学参考。总之，构建GRP78筛选技术平台，具有比较好的科学价值和应用开发价值。