HSCCC+LogP色谱中药吸收成分群整体制备新方法及在川芎研究中的应用

Effective components were the basis of therapeutic drug. The complexity of Traditional Chinese medicine (TCM) and Traditional Chinese medicine formula (TCMF) was the difficulty for clarify effective components and interaction mechanism with modern technologies. Serum Pharmaceutical Chemistry make us realize that successful drug development requires not only optimization of specific and potent recognition by its pharmacodynamics targets, but also efficient delivery to these target sites. To elicit its pharmacological and therapeutic effects, a drug has to cross various cellular barriers by passive. Partition coefficient (LogP) a key determinant in membrane permeability of compounds is useful in estimating the absorption of drug within the body..We first tested integral separation of absorption components from traditional Chinese medicine with HSCCC+LogP chromatography which was characterized by model system i.e. octanol/water two-phase system in high-speed countercurrent chromatography. The main research might include two missions, one is study the accuracy of prediction for absorption components of TCM and the other is the retention characteristic and separation efficiency of model system in the unique exogenic force field with periodic oscillation formed by using rotation and revolution of helical coils of high-speed countercurrent chromatography. According to the oral absorption of TCM and TCMF, predication and separation of absorption components were completed in one step. This research work will helpful to broad the application scope of high-speed countercurrent chromatography on phytochemistry, and promote the development of traditional Chinese medicine materials. Futhermore, HSCCC+LogP chromatography and recycle separating-enriching chromatography were associated to intregal separate absorption components and knockout possible effective components of Rhizoma Chuanxiong for study medicinal material and interaction mechanism.

任何药物对机体起作用，背后都有一个物质基础。中药特别是复方的复杂性是用现代科学语言解读其物质基础和作用机理的困境，时至今日仍缺乏药效成分清楚、机理明确的预防和治疗中药。“血清药物化学”概念的提出，让人们意识到通过吸收成分群研究药效成分群是中药研究的有效手段。本课题在国内外首次提出HSCCC+LogP色谱中药吸收成分群整体制备新方法，该方法最主要的特征是将膜通透性模型体系应用于高速逆流色谱，专注于模型体系对中药吸收成分群准确预测的科学问题，和高速离心力场下固定相保留、分离效率提高等关键技术的研究。从中药或复方成分群在机体中的生物利用度出发，在一次运行中实现中药吸收成分群的预测与整体制备，为研究中药药效物质基础提供新思路和新方法。利用上述新方法整体制备川芎吸收成分群，并应用自主研制的循环制备液相色谱实现精准敲除潜在药效物质or组合，通过动物实验和酶学研究川芎镇静、止痛药效物质及其配伍机理。

中药化合物组成复杂，药效物质不清楚。药效物质作用于机体表现出其活性，必须通过许多生物膜到达作用部位才能实现，膜通透性是药效物质研究的可靠依据。本项目建立了一系列中药成分群分配系数预测模型体系替代正辛醇/水体系，通过分配系数logK的测定，评估中药化合物的膜通透性。中药化合物在正辛醇/异丙醇/0.01M磷酸氢二钾=10:25:25（v/v）体系中的logK与在正辛醇/水体系的logP线性关系良好（logP=1.7638logK-0.0608, R2=0.808），且该体系上、下相极性差较小，适用于较宽极性范围中药化合物的分配系数（-2.1<logP<5.2）测定，弥补正辛醇/水体系极性覆盖的不足。将已建立的中药成分群分配系数预测模型体系和高速逆流色谱结合，建立了中药吸收成分群整体制备新方法。高速离心立场下，正辛醇/异丙醇/0.01M磷酸氢二钾=10:25:25（v/v）体系的流体动力学特征与正辛醇/水体系相反，以该体系下相为固定相，仪器正转，可实现固定相的保留；根据logK与logP、保留体积、流动相流速之间的关系，实现任意logP范围中药化合物的整体制备，为药效物质研究提供新方法，为中药制药研究提供新的思路。再者，针对血浆组成复杂含有高丰度蛋白质干扰物，样本量极少，同时提取极性和弱极性中药化合物困难的问题。本项目建立了基于盐析辅助液液萃取体系异丙醇/4M磷酸氢二钾=1:1(v/v)的血浆样品前处理方法，高效去除高丰度蛋白、脂类、无机盐等杂质，改善血浆样本与LC-MS的兼容性，非特异性富集血浆中极性和弱极性小分子化合物，实现中药入血成分的准确定量分析，为药效物质研究提供技术支撑。此外，将其拓展到血浆、尿液、脑组织等复杂生物样品的内源性代谢物前处理，利用该前处理方法非特异性富集和极性覆盖宽泛的特点，实现内源性类固醇和极性催产素准确定量分析，非靶向代谢组和多肽组研究。基于盐析辅助液液萃取体系的复杂生物样品前处理技术，可为血药浓度检测、内源性代谢物的LC-MS临床高通量检测提供稳定的样品，保证临床检测结果的重复性和准确性。