基于目标成分敲出/敲入的贵重中药麝香药效物质辨识和质量评价方法研究

Quality evaluation has been one of the most important and difficult issues for the modernization of traditional Chinese medicine (TCM), while efficient constituents screening supplies the prerequisite and foundation for quality evaluation. The current quality evaluation pattern for TCM could not assure the safety and efficacy of TCM because this pattern mainly focused on the qualitative and quantitative determination of chemical components which might not be exclusive or active ingredients, while the content limits of index components were set according to the content distribution of several batches of randomly chosen TCM samples. To find a breakthrough of the efficient constituents screening and quality evaluation for TCM, a new pattern based on target component knockout/knock-in strategy imitating gene knockout/transfer was introduced to accurately screen for the key efficent constituents (main efficient components and effect-related components) and explore the interaction properties of them by component knock-out strategy. Further, to set reasonable content limits of these key efficient constituents according to their quantity-efficiacy relationships by component knock-in strategy. All the results will provide some reference for meeting with the requirements of the effect-related, quantitative,accurate and assessable quality evaluation standards. In this research, moschus, an important,expensive, effective and commonly used drug,was chosen as the model TCM. This strategy of target component knockout/knock-in was consistent with the holistic characteristic of TCM, could not only identify the effective components of moschus but also evaluate the quality of it effectively, accurately. It will provide some idea and technical support for the evaluation of other important and expensive TCMs and the exploitation of their artificial substitutes.

中药质量评价是实现中药现代化的关键，而药效物质辨识则是中药质量评价的基础。由于研究思路和方法等的局限，现行的中药质评方法和模式陷入了"难关药效，量而不准，难评难控"的窘境。为了寻求中药药效物质辨识和质评方法的突破，本项目在中医药整体观指引下,在前期初步建立了贵重中药麝香抗炎、抗菌等生物活性评价方法的基础上，借鉴遗传工程中"基因敲出/导入"研究策略，开展基于目标成分敲出/敲入的麝香药效物质辨识和质评方法研究。"敲出"即在谱效关系辅以成分吸收研究基础上，选择性分离目标成分，再通过抗炎、抗菌、调节神经系统功能、兴奋子宫、抗凝血等生物活性检测方法逐步确认关键药效组分；"敲入"即在阴性样品中加入不同比例的目标成分，建立目标成分的量效关系，制订科学合理的药效组分限量范围，最终实现药效物质"找得准，测得准，定得准"的麝香质评目标，亦为贵重中药质量评价和珍稀濒危中药人工代用品研发提供研究思路和技术支持。

中药质量评价是实现中药现代化的关键，而药效物质辨识则是中药质量评价的基础。由于研究思路和方法等的局限，现行的中药质量评价方法和模式陷入了“难关药效，量而不准，难评难控”的窘境。为了寻求中药药效物质辨识和质量评价方法的突破，本项目在中医药的整体观指引下，借鉴遗传工程“基因敲出/导入”研究策略，以贵重中药麝香为研究载体，开展了基于目标成分“敲出/敲入”的中药药效物质辨识与质量评价研究。通过选用与麝香开窍醒神、活血通经、消肿止痛传统功效或临床药效相关的抗心肌细胞和血管内皮细胞氧化损伤、抗凝血酶活性、抗菌（生物热活性）等生物活性评价方法，初步建立了关联麝香药效的体外生物活性评价方法，始终以目标成分与中药麝香整体的等效性比较为准绳；采用“敲出”策略，在谱-效关系研究基础上，选择性地分离目标成分，逐步辨识、确认麝香关键药效组分，实现质量评价指标“找得准、测得准”，通过“敲入”，在阴性样品中有针对性的添加目标成分，建立了目标成分的量效关系，确定了其质量评价标准，制定了科学合理的药效组分限量范围，为后续科学建立麝香质量评价标准及优化天然麝香代用品人工麝香的配方提供了参考，同时也为构建中药药效物质基础及关键药效组分筛选模式提供了研究思路和技术支持。