“等效减毒成分群”导向的红曲制剂质量控制新模式研究

Clinical results indicated that red yeast rice products have preferable comprehensive lipid-lowering ability, also can be used as an alternative for the statin intolerant patients with dyslipidemia. Therefore, this project intends to select Xuezhikang, a red yeast rice preparation having definite clinical curative effect, as the carrier, under the guidance of the working hypothesis “non statin component in red yeast rice product cooperate with statins can play an effect-enhancing and toxicity-reducing effect”, take comprehensive lipid-lowering efficacy and risk of myopathy as evaluation index, based on the overall chemical profile of red yeast rice product, by means of multi level model combined cell, zebrafish and rodent, finding " bioactive equivalent and attenuation combinatorial components" from the complex components which can represent the effect of original product of Xuezhikang, and explore the possible synergy mechanism; on this basis, through the composition -content -efficacy / toxicity data association, multi component characteristics, based on the dose effect weight, using " bioactive equivalent and attenuation combinatorial components " as the labeled amount to establish a new quality control mode for the product of red yeast rice. This project not only provides index combinatorial components for the scientific and reasonable development of quality control for red yeast rice product, but also provides a lead compound combination for the second development and multi-component novel drug discovery of this product.

红曲制剂临床疗效显著，富含他汀成分，且可作为他汀不耐受者的替代性选择（肌病/肝毒低风险）。由于红曲成分复杂，增效减毒成分群不清楚，其质量标准仅以洛伐他汀作为控制指标。基于此，本项目提出“等效减毒成分群”工作假说，即“红曲制剂中存在着一组有限个数、比例明确、含量清楚的成分组合，它既能表征红曲制剂的药效，又具有较低的不良反应风险”。拟以疗效明确的红曲制剂血脂康为研究载体，从中医整体观和中药多成分整合作用为出发点，(1)全面解析其整体化学成分；(2)基于等效成分群反馈筛选，通过不同药理模型，以综合降脂疗效和不良反应发生风险为指标，从红曲制剂血脂康复杂体系中发现其“等效减毒成分群”，探索成分间可能的协同作用机制；(3)通过量效数据关联，构建以“等效减毒成分群”作为红曲制剂有效组分标示量的多成分质量控制新模式。本研究不仅为红曲制剂质量标准提供指标成分群，同时也为多成分组合创新药物的源头研发提供思路。

本项目应用HPLC-MS和GC-MS技术，总结归纳了莫纳可林类、异黄酮类等成分的质谱碎片特征及裂解规律，结合质谱特征数据库/多重过滤方法，从红曲制剂中鉴定化学成分100余个，包括莫纳可林类、十氢萘类、异黄酮类、甾醇类等；大鼠灌胃给药后，从大鼠血浆、尿液和胆汁中共鉴定了化合物109个，包括27个原型成分和82个代谢产物，对红曲制剂的体内代谢途径进行了推测，初步揭示其体内代谢过程特点；采用UF-HPLC-QTOF MS方法，结合分子对接分析，从红曲制剂中筛选得到3个具有胰脂肪酶抑制活性的成分，且存在剂量依赖关系，为非竞争性抑制剂；建立基于主成分敲除策略的二维细胞膜色谱-液相色谱质谱联用方法，从红曲制剂中筛选并鉴定了8个莫纳可林类成分可能具有潜在的心肌保护作用；结合斑马鱼模型，对红曲制剂中莫纳可林类成分、异黄酮类成分、甾醇类成分等进行了安全性评价，结果显示甾醇类对莫纳可林类可能具有潜在的协同降脂减毒作用；开展了红曲制剂的多成分质量评价研究。研究结果在Analytica Chimica Acta、Journal of Chromatography B等杂志上发表SCI论文3篇，申请发明专利1项，培养硕士研究生2人，应邀作国际会议口头报告1次；项目负责人入选2016年度江苏省六大人才高峰计划、2016年度江苏省青蓝工程优秀青年骨干教师，2017年度获得国家优秀青年基金资助。