基于体内动态靶向效应的复方活络效灵丹抗炎物质基础研究

The current approach of developing a single chemical entity targeted on single receptor or signal pathway has not been successfully in term of new medication for treatment of chronic diseases. Huo Luo Xiao Ling Dan (HLXLD) is a classic chinese herbal formula, used for treatment of osteoarthrosis and related diseases, As evidenced by the in vitro binding and functional assays, several anti-inflammatory active molecules have showed significant binding affinity to COX-1/COX-2 or 5-LOX .The objective of this proposal is to establish a multi-targeted (COX-2 + 5-LOX) anti-inflammatory PK-PD model and to examine the effects of bioactive compounds found in HLXLD with anti-inflammatory activities on endogenous biomarker (AA: arachidonic acid). The metabolites of these bioactive compounds will also be monitored for a better understanding of the mechanism of action of HLXLD. This study will not only provide a molecular description of pharmacodynamic material basis of the observed anti-inflammatory in HLXLD, but also validate the approach of multi-targeted and multi-components would stand a better chance to develop effective traditional chinese medicine for prevention and treatment of chronic diseases.

中药复方具有多成分、多靶点之效应, 从而在人类慢性疾病中体现出绝对优于单成分、单靶点药物的疗效优势。本研究以抗骨关节炎中药复方活络效灵丹加减方（HLXLD）为模型方剂，在前期研究基础上，利用作用于体外COX-1/COX-2与5-LOX双靶点的多种"抗炎活性分子"为目标化合物，采用"抗炎活性分子-抗炎药效成分-抗炎靶效成分"逐层推进的策略，应用代谢组学和生物信息学方法构建中药多组分动态的靶向效应PK-PD结合模型，通过获取并"识译"体内与炎症靶点(COX及LOX)密切相关的内源性生物标志物-花生四烯酸代谢性变化，从而在整体动物、分子水平上阐明HLXLD抗炎药效成分体内的动态变化规律，明确HLXLD的药效（靶效）物质基础，多层次揭示复方配伍规律与科学内涵，进而建立起一种基于复方中药抗炎调控靶点和活性分子评价的抗炎有效成分筛选新方法。

本课题体外研究证实活络效灵丹加减方(HLXLD)与炎症靶点COX、LOX之间存在较好的结合和抑制效应。本研究选择了上述双靶点, 基于代谢组学方法，利用UHPLC-MS/MS 方法和手段，对类风湿性关节炎（RA）模型大鼠与正常大鼠体内血浆、组织中花生四烯酸（AA）代谢轮廓进行了定量分析，寻找了AA代谢通路中COX和LOX途径与RA有关的潜在生物标志物，深入探讨了药物干预下机体炎症代谢的关键靶标、作用机制。研究结果表明HLXLD通过双重抑制COX和LOX代谢通路，调节AA代谢轮廓紊乱发挥抗炎作用。鉴于复方中存在一定数量的抑制上述靶点的化合物小分子，以内源性生物标志物-花生四烯酸AA 及其代谢产物代替药效指标以构建PK-PD 模型，动态分析给予HLXLD复方、各单味药及组合后血药浓度和药理效应随时间变化的规律，阐明了 HLXLD抗炎靶效成分以及复方的配伍规律。