基于活血化瘀主效成分群的红花注射液质量评价研究

Carthami Flos (Safflower), a traditional Chinese medicine, is used as an anticoagulation drug in China for a long time. The Safflower injection was prepared by extracts of Carthami Flos for the treatment of occlusive cerebrovascular disease. Although the quality control standard of Safflower injection were improved by Ministry of Health in 2012, the relations between chemical components and pharmacodynamics were still ambiguous. Our researches will focus on the relations between chemical components and pharmacodynamics of Safflower injection, and aim to establish a safe and effective quality control method. Firstly, we will characterize the compounds and its metabolites from Safflower injection and then prepare these compounds; Secondly, screening the main components which possess the ability of anticoagulation is carried on by exploring the association between pharmacodynamics and prototype components and its metabolites; In the end, the quality control standard of Safflower injection will be determined through the content-response relationship, in vivo activity tracing and the contents of main components which possess the ability of anticoagulation. This study intends to promote the quality control of traditional Chinese medicine, gradually changing from "single-component, index components, partial control" to "multi-component, medicinal ingredients, the overall control", and make a revolutionary investigation in quality control of traditional Chinese medicine research.

红花是传统的活血化瘀中药，其现代制剂红花注射液是由红花经水提醇沉等工艺加工而得，临床主要用于治疗闭塞性心脑血管疾病。2012年卫生部药品标准对其质量标准进行了提升，但其质量控制指标与药效的关系仍有待进一步研究。本项目拟以化学成分-药效关联为核心，以质量“安全有效、稳定可控”为目的，对红花注射液开展研究，具体包括：（1）红花注射液化学成分及体内代谢产物表征；（2）以原型成分及代谢产物为对象，利用主效成分群迭代筛选策略，寻找能够代表红花注射液活血化瘀疗效的主效成分群，探索成分间可能的协同整合模式；（3）通过多样本量效关联分析，探讨制剂中主效成分群的含量变化对药效的影响，建立以主效成分群为标示量的红花注射液质量控制新模式。本研究将促进中药质量评价模式逐步从“单成分、指标成分、部分评价”向“多成分、药效成分、整体评价”转变，为中药药效物质及质量控制研究提供借鉴。

本项目通过结合HPLC-QTOF MS建立的定性分析方法，在对醌式查尔酮类、黄酮类及酚酸类等成分质谱特征裂解规律进行研究和总结的基础上，对红花注射液中的化学成分进行了解析，从红花注射液中共鉴定了59个化学成分，包括11个醌式查尔酮、22个黄酮类、14个有机酸类、3个核苷类、5个氨基酸类及4个其它类成分。通过大鼠脑缺血模型对红花注射液的药效进行了评价，其3 ml/kg和6 ml/kg剂量均可明显提高脑缺血大鼠的神经功能评分、减小脑梗死体积，且能持续发挥药效。以N2a、C8和BV2细胞代表中枢细胞，评价了红花注射液对缺氧复氧损伤的保护作用。药代动力学研究显示，与正常大鼠相比，红花注射液中的主要成分羟基红花黄色素A在脑缺血大鼠模型下体内暴露量增高，代谢减缓；结合网络药理学，对与羟基红花黄色素A具有协同作用的成分进行了预测和筛选，提示黄酮类对羟基红花黄色素可能具有潜在的协同作用，并对其中的11个黄酮进行了抗氧化活性评价。此外，开展了红花注射液多成分质量评价研究。发表论文4篇，参与国际交流2次（口头报告1次，墙报交流1次）；培养硕士研究生2人。