基于等效或增效特征挖掘南蛇藤抗肿瘤活性分子群

Traditional Chinese Medicines ingredients are complex and multi-target synergistic, and the active-oriented separation-monomer rescreening model ignores the interaction between components, resulting in the phenomenon of “the molecules do not work”. In the early stage of this project, we have discovered that the ethyl acetate extract of Celastrus orbiculatus (COE) showed significant antitumor activity in vitro and in vivo. Guided by cytotoxic activity screening, 25 compounds were obtained from the COE, but the monomer was ineffective, and the active substances in the COE had not been clarified. This project intends to use reduction method to mine the active molecular groups from COE (COE-BCC) based on the guidance of system theory with the approach of active equivalence or synergism, through the in vivo efficacy evaluation. COE-BCC was characterized by simple composition with controlled quality, and targeted at clear composition and explicable mechanism of action. The following work is proposed to be carried out: (1) Active molecular groups in COE were isolated based on the anti-gastric cancer activity in vivo. (2) In order to elucidate the effective materials of COE, qualitative and quantitative analyses were carried out by using chromatographic separation, spectral identification or chromatography-spectrum combination technologies. (3) After COE-BCC action on tumor-bearing nude mice, the expression level of EMT-related proteins in subcutaneous transplanted tumor tissues was detected by immunohistochemistry. The results of this study will provide the basis for the further research and development of antitumor drugs from Celastrus orbiculatus.

中药成分复杂且多靶点协同作用，活性导向分离-单体复筛模式忽视了成分间的相互作用，导致“只见分子不见效”现象。本项目前期发现南蛇藤茎的乙酸乙酯萃取部位(COE)体内外具有显著的抗肿瘤活性，以体外细胞毒活性为导向，从COE中获得25个单体，但单体活性不佳，COE的抗肿瘤活性物质一直未得到阐明。本项目拟在系统论指导下，采用动物体内药效评价模式，以活性等效或增效为原则，以组成简约、质量可控为特点，以成分明晰、作用机制可阐释为目标，运用还原法挖掘COE中的活性分子群。拟开展以下工作：(1)以体内抗胃癌活性为导向，分离COE中活性分子群 (COE-BCC)。(2)运用色谱分离、光谱鉴定或色谱-光谱联用等技术，定性定量COE-BCC，阐明其药效物质基础。(3) COE-BCC作用荷瘤裸鼠后，采用免疫组化法检测皮下移植瘤组织中与EMT相关的蛋白表达水平。该结果将为南蛇藤抗肿瘤药物研究与开发提供理论依据。

本项目在系统论指导下，采用动物体内药效评价模式，以活性等效或增效为原则，以组成简约、质量可控为特点，以成分明晰、作用机制可阐释为目标，运用还原法挖掘COE中的活性分子群。具体开展工作如下：以体内抗胃癌活性为导向，分离COE中活性分子群 (COE-BCC)；运用色谱分离、光谱鉴定或色谱-光谱联用等技术，定性定量COE-BCC，阐明其药效物质基础；COE-BCC作用荷瘤裸鼠后，采用免疫组化法检测皮下移植瘤组织中的EMT相关蛋白的表达水平。实验结果表明COE-BCC为二萜和三萜分子群，COE-BCC通过下调皮下移植瘤组织中Vimentin、N-cadherin、HSP27、NF-κB p65和Snail蛋白的表达，上调E-cadherin的表达，抑制胃癌细EMT的发生，进而抑制胃癌的发展。本项目阐明了COE抗胃癌的药效物质基础和初步作用机理，为南蛇藤用于抗胃癌药物开发提供重要科学依据。