基于目标成分敲除/敲入的中药五加皮抗炎药效物质辨识与质量控制研究

The quality control of Chinese materia medica (CMM) is very crucial in the modernization of CMM, and screening of biologically active components of CMM plays a key role in this process. Although various pharmacological activities of root bark of Acanthopanax gracilistylus (AG) were revealed by bioactive tests in vitro or in vivo, the active components (ACS) responsible for expelling wind-damp have not been elucidated, and its quality control standard (QCS) is also imperfect. On the basis of our previous bioactive investigation and LC-MS (liquid chromatography linked with mass spectrometry) analyses of active parts, in this project, we will make a systematic study of ACS and QCS of AG by using the theory of target components knock-out/knock-in research mode. The current proposal aims to firstly knock out main components from AG and then comprehensively evaluate anti-inflammatory activities of target components, non-target component fractions and crude extract of AG in vitro and in vivo. The bioactive of partial samples will be throughout compared with the total extracts of AG, and the key efficient components of anti-inflammation of AG might be recognized, and the dose-effect relationship of ACS might be also revealed by the strategy of knock-in mode. Finally, the current project would facilitate identification of main active components and quality evaluation of AG, and also provide some research ideas and references for investigating the main active components, quality control and impelling modernization of CMM.

中药质量控制是制约中药现代化发展的关键问题之一，而中药药效物质辨识则是支撑中药质量控制的重要基石。目前，尽管中药五加皮在体内外试验中显示了广泛而显著的药理活性，但其主要临床功效祛风湿的关键药效物质仍未阐明，其质量控制标准仍不完善。本项目采用“基于目标成分敲除/敲入的中药药效物质辨识与质量控制研究模式”，在前期体内外活性研究及关键活性部位LC-MS分析的基础上，选择性地“敲除”其主要目标成分，并始终以目标成分与中药整体的等效性比较为准绳，逐步辨识、确认其抗炎关键药效组分；通过在阴性样品中有针对性地“敲入”目标成分，建立目标成分的量效关系，制订科学合理的质量控制限量范围，实现质控指标“关联药效”。本项目的研究结果，将为科学全面评价五加皮抗炎物质基础提供科学数据，为完善五加皮药材质量标准提供参考，同时也为中药药效物质辨识、中药质量控制和中药现代化研究提供一些思路和借鉴。

中药五加皮是临床常用的祛风除湿类中药，疗效显著；但其主要临床功效祛风湿的关键药效物质仍不明确，其质量控制标准仍不完善。为了辨识五加皮功效物质，从而为其质量控制提供重要基石，本项目基于中医药的整体观，采用“目标成分敲除/敲入的中药药效物质辨识与质量控制研究模式”，对中药五加皮关键功效物质进行了研究。构建了关联五加皮主要临床功效的抗炎活性评价方法，在此基础上，始终以活性评价为准绳，开展了五加皮提取物抗炎药效物质确认与辨识，明确了抗炎活性部位，并采用LC-IT-TOF-MS/MS技术，结合各种色谱分离技术，对关键活性部位进行了有针对性的分析和分离工作，获得了其主要活性成分；并进一步采用各种波谱技术结合量子化学计算的方法确定了活性成分的结构，其中6个成分为新化合物，包含3个比较稀有的天然产物和1个具有新奇骨架类型的化合物，并首次从五加皮中获得了倍半萜和单萜类成分。生物活性评价结果表明，5个倍半萜类成分具有显著的体外抗炎作用，这将有助于拓展对五加皮功效物质的认识。本项目研究结果初步明确了五加皮抗炎药效物质，为五加皮临床应用提供了支持，为后续制定科学合理的质量标准奠定了良好基础，为五加皮的进一步开发和应用提供了参考，同时也为其它中药药效物质辨识及关键药效组分筛选提供了研究思路和技术示范。