中药透皮成分快速筛选新方法在外敷梅花点舌丸抗CP/CPPS药效成分研究中的应用

How to rapidly study the pharmacodynamic material basis of externally-applied traditional Chinese medicine (EA-TCM) has been one of the challenging issues in the field of traditional Chinese medicine so far. The previous studies from the applicant and others suggested that human skin could exert a screening effect on the chemical components of EA-TCM. Based on an interesting strategy to find out the active components of EA-TCM through its skin-permeating components, that is, the screening mode “total chemical components - skin-permeating components - activity evaluation - active components”, the applicant proposed a novel method to study the pharmacodynamic material basis of EA-TCM based on our constructed linear free-energy relationship (LFER) equation that is used to predict the skin permeabilities (log Kp) of compounds. In this study, externally-applied Mei-Hua-Dian-She-Wan (MHDSW) treating chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) will be taken as a study example. The potential skin-permeating components of MHDSW will be screened out using the LFER equation, by which the log Kp values of all its chemical components are calculated. The chemical components of MHDSW permeating across human skin will be collected via in vitro and in vivo skin permeation experiments, and further analyzed by HPLC/GC-HR-MS/MS technique to verify the presence of the potential skin-permeating components. As a result, the skin-permeating components of MHDSW can be rapidly identified. The skin-permeating components will be tested on a rat model of experimental autoimmune prostatitis as CP/CPPS model to screen out the therapeutic components. This study is expected to establish a rapid and accurate method to elucidate the pharmacodynamic material basis of EA-TCM, which will help us understand the pharmacological nature of EA-TCM.

如何快速研究外敷中药药效物质基础是中药研究领域的难题之一。现有研究表明，人体皮肤对外敷中药化学成分有明显的筛选作用。申请人采用基于透皮成分的外敷中药药效物质筛选思路，即原药全成分-透皮成分群-药效评价-活性成分组的筛选模式，结合前期研究，提出基于皮肤渗透系数LFER方程预测透皮成分的外敷中药药效物质基础研究新方法。本项目将以梅花点舌丸外敷治疗慢性非细菌性前列腺炎（CP/CPPS）为研究模型，利用已建立的LFER方程预测梅花点舌丸各化学成分的皮肤渗透系数，筛选出潜在透皮成分。并通过体内外皮肤渗透实验收集其透皮成分，利用液相/气相色谱-质谱技术验证潜在透皮成分，建立梅花点舌丸透皮成分群。进而利用大鼠CP/CPPS模型，从透皮成分群中筛选出药效成分组，以阐明外敷梅花点舌丸抗CP/CPPS药效物质基础。本项目的实施，有望建立一个快速、准确的外敷中药药效物质筛选新方法，有助于阐释外敷中药的科学内涵。

如何高效研究外敷中药药效物质基础是中药研究领域的难题之一。前期研究表明，人体皮肤对外敷中药化学成分有明显的筛选作用。申请人采用基于透皮成分的外敷中药药效物质筛选思路，即原药全成分-透皮成分群-药效评价-活性成分组的筛选模式，结合已有研究，提出基于线性自由能关系（LFER）方程预测透皮成分的外敷中药药效物质基础研究方法。本研究以吴茱萸和金银花为模型中药，探讨这一新方法的可行性。我们利用LFER方程分别计算吴茱萸和金银花化学成分的皮肤渗透系数（log Kp），评估其皮肤渗透能力。通过体外皮肤渗透实验分别收集吴茱萸和金银花的透皮成分，并使用高效液相色谱-高分辨串联质谱进行分析鉴定。研究中药透皮成分的log Kp依赖性，同时探讨透皮成分的药理活性与临床疗效之间的联系。研究结果表明，LFER方程可以快速筛选吴茱萸和金银花的潜在透皮成分。同时，log Kp ≥ ‒7.0被建议作为外敷中药潜在透皮成分的筛选标准。非常有意义的是，吴茱萸的主要透皮成分很好地解释吴茱萸外敷抗高血压作用。因此，LFER方程可以快速筛选出外敷中药潜在透皮成分，缩小有效成分范围，加速药效物质基础研究，有助于外敷中药的现代化开发和利用。