基于血清谱效学和PK-PD模型的苗药云实皮抗炎药效物质基础研究

Caesalpinia decapetala（Roxb.）Alston, a Miao national herb in Guizhou, is commonly used for treatment and prevention of bronchitis and influenza, and a number of listed ethnic preparations contain C. decapetala. Although its chemical components and pharmacological activity have been reported, its anti-inflammatory therapeutic material basis and mechanism remain unclear. In our previous study, it was the first time to found that the water soluble fraction, of which 11 were found from this species for the first time, also had a significant effect on the inflammatory model. Based on the previous systematic research of chemical components, pharmacological action and effective soluble fraction, the project will focus on those migrating into the blood by means of serum spectrum-activity analysis and the fingerprint of UPLC/Q-TOF MS technology. Furthermore, the potential relevance between these constituents and pharmacological action will be put into the study to explore the anti-inflammation ones, using inflammatory models. Thereafter, the serial research will be conducted on ADME and anti-inflammation test by PK markers, using animal disease model and cell model, so as to build the PK-PD model which may demonstrate the action between the multicomponents and the pharmacological activities (Aiming at the relevance of the Concentration, Time and Effectiveness). Through the metabolism fate study of the anti-inflammation effective constituents and their dynamic transformation in vivo, it can help find out the therapeutic material basis in the herb and to lay a theoretical and experimental foundation for further development and application of the medicinal plant.

云实皮为贵州苗族常用药材，对于支气管炎、流行性感冒的治疗有独特疗效，已有多个民族药品种上市。课题组首次研究发现云实皮乙醇提取物的水溶性部位具有显著的抗炎作用（首次分离得到11个化合物），但其药效物质基础、作用机制及体内过程尚不明确。本研究旨在以云实皮有效组分为研究对象，以血清谱效学为手段，以UPLC/Q-TOF MS技术建立指纹图谱，全面分析云实皮入血成分，基于炎症模型探讨体内移行成分与抗炎活性的相关性，从病理状态和细胞药代动力学的角度，建立PK-PD模型，开展云实皮在炎症疾病动物模型和细胞模型下的药代动力学和抗炎作用相关性研究（即药物浓度-时间-效应关系），揭示其抗炎作用的效应成分及体内动态变化规律，最终阐明云实皮抗炎作用的物质基础，为其深层次开发利用奠定理论和试验基础。

云实皮主产贵州，为贵州苗族常用药材，具有袪风除湿，解毒消肿的功效，具有较强地域特色资源优势和发展潜力。 但云实皮的药效物质基础、作用机制以及体内过程研究尚属空白， 难以保证云实皮及其相关产品的质量和疗效， 并已成为制约云实皮产品升级换代和产业链做大做强的瓶颈问题。本项目通过云实皮化学成分研究分离得到4对二苯并氧辛类新化合物；采用UHPLC/Q Exactive Plus MS技术结合Compound Discoverer 3.1数据分析软件对云实皮化学成分和体内的移行吸收入血成分进行全面表征。从云实皮中分析鉴定了21个化学成分，主要包括原苏木素类、黄酮类、萜类、查耳酮类等。在大鼠血清中共检测到53个体内移行成分，其中8个成分为原型成分，21个为原型成分的代谢产物，通过抗炎效应成分血清谱效关系分析确定了PK markers。完成了正常和AA大鼠单次和多次口服给予云实皮提取物后主要活性成分的药动学特征研究及其差异性分析，发现3个活性成分在AA大鼠体内的吸收量增高，清除率降低，并研究建立了PK-PD结合模型，提示药效指标IL-1β、RF、IL-6与云实皮活性成分的血药浓度存在良好相关性。基于LPS诱导RAW264.7炎症细胞模型探讨了云实皮提取物的抗炎作用机制，云实皮能抑制p65入核、TNF-α蛋白的表达和AKT、IκB、P65蛋白的磷酸化以及AKT通路的激活，从而发挥抗炎作用，表明其抗炎作用机制与TNF-NF-κB-AKT信号通路有关。从细胞药代动力学的角度建立了云实皮提取物5个主要活性成分的PK-PD结合模型，研究提示云实皮活性成分发挥抗炎的作用可能与下调炎症细胞中NO和TNF-α分泌有关， NO和TNF-α可能为云实皮提取物作用于炎症相关作用靶点。建立了基于云实皮药效物质的多指标质量控制新方法。上述研究为云实皮创新药物研制及药材资源深度开发提供了科学依据。