基于功效成分群的中成药生产全过程控制体系

肖 伟（中国药理学会中药与天然药物药理专业委员会副主任委员，江苏康缘药业股份有限公司研究员级高级工程师，中药制药过程新技术国家重点实验室，主要从事中药新药研究与开发。

曹 亮（世界中医药学会联合会网络药理学专业委员会常务理事，江苏康缘药业股份有限公司高级工程师，主要从事中药新药研究与开发。

中药标准化是国家重大战略需求，已被纳入《国家中长期科学和技术发展规划纲要》。目前，中成药标准现状与国际药品标准水平存在较大差距，严重阻碍了中成药的现代化、国际化进程，突出表现在两大方面，一是质控成分相对单一，多与功效关联性不强，难以保障整体功效；二是质控成分无明确质控范围（有下控制限而无上控制限），生产过程中缺乏控制，产品批间波动大，难以保证疗效稳定。导致上述现状的重要原因有：①缺乏系统的功效成分发现模式及功效成分在中成药系统下确定量效范围的方法；②在中成药生产中缺乏多成分整体控制并稳定传递的技术体系。基于此，我们首先突破中药功效成分快速确证技术，结合中成药化学成分规模化、快速分离制备方法，集成高通量-高内涵活性筛选、体内代谢和整体药效验证，构建起预测-制备-验证-聚焦-确证的中药功效成分发现模式；其次，整合生物通路的定量计算方法和分子印迹技术，引入中药多成分-多功效的量-效范围研究来明确中药功效成分的“质量控制窗”，解决中成药质控范围难以确定的问题；然后，建立基于质量源于设计（QbD）理念的工艺辨析优化技术和过程融合指纹图谱技术，集成自控和在线检测技术，形成生产过程中功效成分群的“各工艺点的点-点一致”、“各工序段的段-段一致”、“各生产批次的批-批一致”的质量一致性控制体系，突破中成药质量均一和疗效稳定难题。【According to the Outline of the National Program for Long-and Medium-Term Scientific and Technological Development,we will encourage the standardization of TCM.Moreover,the reality is that few compounds have their quality stan⁃dard controlled and the compounds are not attrib⁃uted to a certain efficacy in the most situations,and that the upper limit of the compounds is not defined,thus having negative influence on the process of TCM and posing an obstacle to global⁃ization of related trade and the standardization of TCM.To address these issues,some strategies have been initiated to discover active compounds and to ascertain the optimal window of the compounds at system levels.Some modern tech⁃nologies are employed to monitor the change of active compounds ranging from Chinese herbal medicine to preparations.Firstly,a platform is set up by integrating virtual screening,preparative chromatography,high throughputscreening,high content screening,pharamcokinetic analysis and animal models.The platform can then prepare a large number of hits and verify the effective compounds in TCM.Secondly,the reasonable range of effective compounds is defined in quality control for TCM via molecular imprinting and quantitative modeling based on pathway(QMBP).Finally,according to the concept of quality by design(QBD),a control system is built to ensure the stability from a parameter to a procedure in the production process forevery batch by combining fingerprint chromatogram,and on-line/off-line real-time quantitative monitoring.】

近年在上述研究思路的指导下，对源于张仲景的经典方—桂枝茯苓方的中药大品种桂枝茯苓胶囊进行了基于功效成分群的全过程质量控制体系进行了研究，利用具有自主知识产权的中药生物活性信息预测分析和制备工业色谱制备164个潜在功效成分的百克级化合物库，通过高通量-高内涵平台验证26个功效成分，经体内代谢研究和整体动物评价确证了制剂治疗原发性痛经、盆腔炎和子宫肌瘤的15个功效成分包括芍药苷、丹皮酚、去氢茯苓酸等；借助溶胶-凝胶法制备分子印迹聚合物分别敲除制剂中相应功效成分，整合生物通路的定量计算方法，确定各个功效成分的整体量效关系，进而设定功效成分的“质量控制窗”作为质量标准制订依据；以功效成分传递波动性为指标对工艺全程进行质量风险分析，确定影响质量一致性的关键工序段和工艺环节，并进一步辨识各工艺参数与质量波动的相关性明确法定工艺参数，之后采用近红外在线监测、离线快速分析，自动化控制等技术对法定/非法定工艺参数、关键工序段和工艺环节进行布局设计，达到“点-点控制”和“段-段控制”，最后建立从原料、中间体到成品的基于功效成分质控窗的定量指纹图谱检测生产的每批制剂，累积分析质量的均一性和疗效的稳定性，从而达到“批-批一致”。通过近3年共多批次桂枝茯苓胶囊大生产的验证，成品中各功效成分含量及指纹图谱相似度等质控指标批次间RSD%由8.3%降至3.6%，明显提高了质量一致性，保障临床疗效稳定，实现了“批-批一致”，也验证了该体系的科学可行。【Take Guizhi Fuling capsules for example.The change of the effective compounds in the process of production was explored to set up a perfect quality system.The hits,which were fished on the basis of the indication from Traditional Chinese Medicine Network Phar⁃macology Intelligent Information Platform,were separated up to hundreds of grams by preparative chromatography.Among the compounds,there were 26 active ingredients by high throughput screening and high content screening,15 of which(including paeoniflorin,paeonol,and dehy⁃dropachymic acid)were verified by identifying the components in plasma and evaluating the effect in some animal models relating to dysmenor⁃rhea,pelvic inflammatory disease,and endome⁃triosis.Then,the molecularly imprinted polymers(MIPs)of the effective compounds were synthe⁃tized via a sol-gel coating process.Each sample without a compound in Guizhi Fuling capsules was prepared by MIPs,and the dose-response curve was computed and tested by QMBP and in vivo for the effective compounds at the system level.The quality standard was performed in Guizhi Fuling capsules according to the optimal windows of the dose-response curve.During the produc⁃tion of Guizhi Fuling capsules,the key procedure and the parameters closely related to the uniform quality were defined by analyzing the fluctua⁃tions of the content of the effective compounds,and then were monitored by on-line/off-line nearinfrared spectroscopy and automatic control system.Finally,HPLC fingerprints were developed to quantify the effective compounds from Chinese crude drugs,intermediate products to prepara⁃tion for every batch,and the variation of the effective compounds was reduced from 8.3%to 3.6%.The data from three years showed that the uniformity of quality was improved,and that the products had a stable therapeutic effect in clinic.It proved that the strategy was feasible for total quality control of TCM.】