Herroon Mackenzie K, Sharma Rajgopal, Rajagurubandara Erandi, Turro Claudia, Kodanko Jeremy J, Podgorski Izabela
Biol Chem. 2016 Jun 1;397(6):571-82. doi: 10.1515/hsz-2015-0274.
Collagenolytic activity of cathepsin K is important for many physiological and pathological processes including osteoclast-mediated bone degradation, macrophage function and fibroblast-mediated matrix remodeling. Here, we report application of a light-activated inhibitor for controlling activity of cathepsin K in a 3D functional imaging assay. Using prostate carcinoma cell line engineered to overexpress cathepsin K, we demonstrate the utility of the proteolytic assay in living tumor spheroids for the evaluation and quantification of the inhibitor effects on cathepsin K-mediated collagen I degradation. Importantly, we also show that utilizing the ruthenium-caged version of a potent nitrile cathepsin K inhibitor (4), cis-Ru(bpy)2(4)22 (5), offers significant advantage in terms of effective concentration of the inhibitor and especially its light-activated control in the 3D assay. Our results suggest that light activation provides a suitable, attractive approach for spatial and temporal control of proteolytic activity, which remains a critical, unmet need in treatment of human diseases, especially cancer.
组织蛋白酶K的胶原酶活性对许多生理和病理过程都很重要，包括破骨细胞介导的骨降解、巨噬细胞功能和成纤维细胞介导的基质重塑。在此，我们报告了一种光激活抑制剂在三维功能成像分析中用于控制组织蛋白酶K活性的应用。利用经过基因工程改造以过度表达组织蛋白酶K的前列腺癌细胞系，我们证明了蛋白水解分析在活肿瘤球体中用于评估和定量抑制剂对组织蛋白酶K介导的I型胶原降解的作用。重要的是，我们还表明，使用一种强效腈类组织蛋白酶K抑制剂（4）的钌笼合物形式，即顺式-Ru(bpy)2(4)22（5），在抑制剂的有效浓度方面，尤其是在三维分析中的光激活控制方面具有显著优势。我们的结果表明，光激活为蛋白水解活性的时空控制提供了一种合适且有吸引力的方法，而这在人类疾病尤其是癌症的治疗中仍然是一个关键的、未得到满足的需求。
