基于分子烙印技术的波罗蜜属植物中异戊烯基黄酮类组织蛋白酶K抑制剂的快速发现

Osteoporosis (OP) is a common and frequently occurring illness which threatens the health of the aged heavily. However, no ideal drugs have been developed to cure it so far. Recently cathepsin K (CatK) has been confirmed to be a certain and effective target for OP and CatK inhibitors is reported to be efficacy for the treatment of OP. Previous investigation and the claimer's data indicate that the isoprenylated flavonoids from Artocarpus plants are an important natural source of CatK inhibitor. However, the efficient dicover of CatK inhibitor from extracts of Artocarpus plants is difficult by employing conventional phytochemical methods. In view of this, this project is designed to perform a series of studies as following: The preparation of AA-MIP with artonin A, an isoprenylated flavonoids CatK inhibitor isolated from an Artocarpus plant in claimer's previous study, as the template molecular, followed by the establish of AA-MIP chromatogram system; The recognization and separation of isoprenylated flavonoids from five Artocarpus plants by application of AA-MIP chromatogram, followed by further isolation and structure elucidation of compound; Studies on the relevance of the AA-MIP chromatographic retention characteristics of compound with their CatK selective inhibition activity. This project aims to establish a new AA-MIP chromatogram technique system for the efficient discovering of isoprenylated flavonoids CatK inhibitor from Artocarpus. Meanwhile, the execution of this project will provide lead compounds with selectivity inhibitory activity against CatK for the R & D of new drugs used for the treatment of OP.

骨质疏松症(OP)是严重威胁中老年人健康的常见、多发病，目前尚无理想的治疗药物。组织蛋白酶K(CatK)近年来被确认为治疗OP的新靶点，其抑制剂对OP有明显的治疗效果。文献报道及申请者前期的研究提示，波罗蜜属植物中的异戊烯基黄酮类成分为天然CatK抑制剂的重要来源，然而目前传统的植物化学方法难以高效快速的获得。基于此，本项目拟进行以下研究：以前期分离的异戊烯基黄酮类CatK抑制剂-artonin A为模板分子制备分子烙印聚合物(AA-MIP)，并以其为固定相建立AA-MIP色谱；以AA-MIP色谱对5种波罗蜜属植物的异戊烯基黄酮类成分进行识别分离，纯化并鉴定单体结构；研究单体的色谱保留行为与其活性相关性规律。本项目旨在构建快速识别、分离异戊烯基黄酮类CatK抑制剂的AA-MIP色谱新技术，获得具有CatK选择性抑制活性的异戊烯基黄酮类成分，为新型抗OP药物研究提供先导化合物。

骨质疏松症(OP)是严重威胁中老年人健康的常见、多发病，目前尚无理想的治疗药物。组织蛋白酶K(CatK)近年来被确认为治疗OP的新靶点，其抑制剂对OP有明显的治疗效果。文献报道及申请者前期的研究提示，波罗蜜属植物中的异戊烯基黄酮类成分为天然CatK抑制剂的重要来源，然而目前传统的植物化学方法难以高效快速的获得。基于此，本项目进行了以下研究：○1 优化了以artonin A为模板的分子烙印聚合物(AA-MIP)的制备工艺参数：最优功能单体为4-vp，致孔剂为丙酮，交联剂为EDMA，最佳物料比为n(AA):n(4-Vp):n(EDMA)=l:5:25；○2 通过吸附动力分析、吸附等温线分析表征了AA-MIP聚合物的吸附性能，scatchard分析表明AA-MIP对模板分子有单一的作用位点；○3 建立了AA-MIP柱色谱体系，利用其对两种波罗蜜属植物-波罗蜜和二色波罗蜜的总黄酮进行了识别富集，并建立了两个富集物的HPLC指纹图谱；○4 HPLC指纹图谱指导下的化学成分研究从两个总黄酮部位分离获得了101个单体成分。采用光谱法鉴定了所有单体的化学结构，包括异戊烯基黄酮40个，异戊烯基2-芳基苯并呋喃20个、异戊烯基二苯乙烯13个及其它酚性化合物28个；○5 评价了所分离单体对CatK的抑制活性，其中的5个异戊烯基黄酮对CatK具有选择性抑制活性，并测定了它们对CatK的解离常数Kd。本项目探讨了一种快速发现CatK抑制剂的分子烙印色谱技术，为新型抗OP药物研究提供了可能的先导化合物。