基于肠吸收谱-效相关的吴茱萸汤治疗偏头痛有效组分辨识优化研究

As an effective and classic TCM formulation for the treatment of migraine, the pharmacological active ingredients and their ratios in Wuzhuyu-tang siginificantly affect its clinical effects. Therefore, extensive investigations are urgently necessary to be further performed in detail. New strategy, i.e. correlation analysis of intestinal absorption spectra-pharmacological effects was firstly introduced here to solve currently facing difficulties: complicated isolation and characterization of chemical components encountered in compound spectra-effects relationship study, and more complicated PK-PD problems in spectra-effects relationship study of ingredients absorbed into blood. Firstly, the key ingredients of interest will be assigned in those components absorbed into averted intestine sac and intestinal segments in situ using multi-indexes related to migraine. Secondly, the pharmacological active ingredients in vitro will be sought out through relating the chemical information of Wuzhuyu-tang to that of the key attention ingredients. Thirdly, the ratios of pharmacological active ingredients are optimized with UDM-SVMANM-EMOO triplex-process optimization method, which adapt to the character of Traditional Chinese Medicinal Compounds. Finally, the pharmacological active ingredients and the optimization ratios of Wuzhuyu-tang in treating migraine will be identified and validated. Furthermore, the pharmacokinetic and brain tissue distribution behaviors of the optimized samples in vivo will be compared with those from original samples, in order to improve the therapeutic effects of Wuzhuyu-tang and confirm present new rapid-discovery and optimization strategy of pharmacological active components suitable for combination formulations of TCM.

吴茱萸汤是治疗偏头痛的有效经典方剂，有效组分的明确对其疗效有重要影响，亟待深入研究。本课题针对目前常用研究方法中存在的难点提出解决新思路，即以肠吸收谱-效相关为切入点，降低药物谱效相关需对化学成分全面分离、表征的难度，减少吸收入血成分与药效相关面临的PK-PD复杂问题。通过在体单向肠灌流吸收、离体外翻肠囊吸收试验，以及药效学试验，建立"相对有限的吸收成分"与多个偏头痛相关药效指标改善之间的关系，采用STEPCARD法筛选吸收成分中的"重点关注组分"，以及汤剂中影响"重点关注组分"的体外"有限有效组分"。针对筛选出的有效组分，采用适应复方组分优化特点的UDM-SVMANM-EMOO三联法，寻找改善药效指标的优化组分剂量与比例并进行药效学验证，比较优化前后样品体内药物动力学特征与脑组织分布探究机理，以提高吴茱萸汤治疗偏头痛疗效，验证本课题建立的更适合中药复方特点的有效组分快速发现和优化方法。

吴茱萸汤是治疗偏头痛的有效经典方剂，有效组分的明确对其疗效有重要影响，亟待深入研究。本课题针对目前常用复方药效物质基础研究方法中存在的难点提出解决新思路，即以肠吸收谱-效相关为切入点，降低药物谱效相关需对化学成分全面分离、表征的难度，减少吸收入血成分与药效相关面临的PK-PD复杂问题。通过在体单向肠灌流吸收、离体外翻肠囊吸收试验，以及药效学试验，建立“相对有限的吸收成分”与多个偏头痛相关药效指标改善之间的关系，采用熵值加权-偏最小二乘回归法筛选吸收成分中的“重点关注组分”，以及汤剂中影响“重点关注组分”的体外“有限有效组分”。针对筛选出的有效组分，采用适应复方组分优化特点的均匀设计-熵值加权/总评归一值-逐步回归/偏最小二乘回归三联法，寻找改善药效指标的优化组分剂量与比例并进行药效学验证，并比较优化前后样品中“重点关注组分”的在模型动物上的单向在体肠灌流吸收差异、体内药物动力学特征与脑组织分布差异，探究机理，以提高吴茱萸汤治疗偏头痛疗效，验证本课题建立的更适合中药复方特点的有效组分快速发现和优化方法。. 结果表明，吴茱萸汤治疗不同偏头痛动物模型的重点关注组分不同。对于硝酸甘油偏头痛大鼠模型，汤中成分的含量（mg•mL-1）为Rb1=1.4550，Rv=0.0090，Ev=0.5650， Ru=0.0810，Rg1=0.8710，Li=0.1780，6-Gi=0.1590，吴茱萸汤改善偏头痛的综合药效最好。而Ev、Rb1、6-Gi、Ru的肠吸收量增加、Ev的体内滞留时间延长、Rb1的入血吸收量增加可能是其疗效好的原因。对于利血平偏头痛小鼠模型，汤中成分的含量（mg•mL-1）为Ev=0.3120，Rb1=1.054，Rg1=1.2336，Re=0.0912，Li=0.3338，6-Gi=0.2986，吴茱萸汤改善偏头痛的综合药效最好。Ev、Rb1、6-Gi的肠吸收量增加、Ev、Rb1、Rg1的入血吸收量增加可能是其疗效好的原因。