组织蛋白酶K抑制剂调控的骨代谢过程及相关动态机制和分子机理研究

Osteoporosis is a debilitating disease when bone resorption exceeds bone formation and it seriously endangers the health of the elderly in global. Cathepsin K (Cath K) has been shown to play a key role in bone metabolism, degrading bone matrix in bone resorption and regulating the bone mineralization in bone formation. Therefore, by using the already achieved highly potent and selective azapeptides nitrile Cath K inhibitors as sensors to regulate the activity of cathepsin K, multiple chemical and biological combined methods and technologies will be performed to the fields as: (1) Construction the cell models of osteoblasts and osteoclasts, and using them to monitor bone metabolism processes as bone mineralization and resorption under the contral of cathepsin K and its inhibitors. The purpose is to screen out inhibitors having high selectivity on Cath K in osteoclasts, and essentially showing promotion of bone mineralization in osteoblasts; (2) Building SaOS-2 derived extracellular matrix (ECM) as resorption model, and chicken embryo derived matrix vesicle (MVs) as minaliztion model to realize the in situ monitoring and kinetics measuring of the two processes, in final to reveal the involved molecule mechanism of bone metabolism and, essentailly, the reason induce promotion of bone mineralization; (3) Revealing the key points and other regulation essence of bone metabolism, providing new research highlight and foundamental basis for the development of new drug candidates, in targeting cathepsin K for the treatment of osteoporosis and other bone diseases.

骨质疏松症是骨形成与骨吸收之间动态平衡失衡后产生的一种骨骼疾病，它严重危害着全球中老年人的健康。组织蛋白酶K (Cath K）是参与骨代谢的主要酶：它既在骨吸收过程中参与骨基质降解,又和骨形成过程中的骨矿化密切相关。因此，我们将以肼腈类Cath K抑制剂为调控探针，拟利用化学和生物学多种技术相结合方法, 通过对Cath K活性的调控开展如下研究： (1)建立成骨细胞和破骨细胞研究模型，在细胞层次上研究抑制剂对骨吸收和骨矿化过程的调控和影响，以优化出既具有细胞水平高选择性又能促骨矿化的小分子抑制剂；（2）分别建立可用于体外原位跟踪检测的骨吸收和骨矿化仿生物模型，以实现对相关动力学机制的原位检测和在分子水平上揭示相关的分子机理；(3)揭示抑制剂具有抑骨吸收和促骨矿化协同效应的关键因素和其它可能调控因子，为开发以Cath K为靶标的骨疾病新药提供新的研究思路和理论基础。

骨质疏松症是骨形成与骨吸收之间动态平衡失衡后产生的一种骨骼疾病，它严重危害着全球中老年人的健康。组织蛋白酶K (Cath K）是参与骨代谢的主要酶：它既在骨吸收过程中参与骨基质降解,又与骨形成过程中的骨矿化密切相关。我们以肼腈类Cath K抑制剂为调控探针，利用多种化学和生物学技术相结合方法,开展了如下研究： (1) 利用小鼠骨髓提取液分别建立了成骨细胞和破骨细胞研究模型;基于此系统研究了系列I抑制剂对骨代谢过程的调控和影响; 揭示了其中的分子机理, 并优化出2个高效小分子抑制剂CKI-8与CKI-13；（2）建立了可用于体外原位跟踪检测骨吸收和骨矿化进程的骨片模型，系统地研究了相关动力学机制、并在分子水平上揭示对应的分子机理；牛腿骨骨片的骨吸收检测模型结果表明，CKI-8与CKI-13不仅能够抑制骨吸收效应，还能对细胞在骨表面的迁移产生抑制作用。 (3) 揭示了肼腈类Cath K抑制剂构-效关系本质，并基于此重新设计合成了系列II和III新型抑制剂，为开发以Cath K为靶标的骨疾病新药提供新的研究思路和理论基础。