高通量赖氨酰羟化酶（LH）测定法的建立及 LH2 小分子抑制剂的鉴定。
Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2.
机构信息
1 Targeted Therapeutic Drug Discovery and Development Program, College of Pharmacy, The University of Texas at Austin, TX, USA.
2 Department of Thoracic Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
出版信息
SLAS Discov. 2019 Apr;24(4):484-491. doi: 10.1177/2472555218817057. Epub 2018 Dec 27.
Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.
赖氨酰羟化酶-2（LH2）催化胶原蛋白末端肽赖氨酸残基的羟化，形成稳定的胶原交联，连接胶原分子并稳定细胞外基质。研究报道，在形成和稳定羟赖氨酸醛衍生的胶原交联（HLCCs）中，LH2 的水平较高，导致某些组织中的纤维化和癌症转移。鉴定针对 LH2 活性的小分子抑制剂需要一个强大且合适的测定系统，而目前这方面的系统还很缺乏。因此，尽管 LH2 是这些疾病的一个有前途的靶点，但针对 LH2 的小分子抑制剂尚未被报道。因此，我们开发了一种基于发光的策略来监测 LH 活性，并通过对大约 65000 种化合物对 LH2 的筛选，验证了其识别新抑制剂的能力。使用针对 mimiviral L230 的相同 LH 测定法对主要命中化合物进行了确认。新开发的 LH 测定法具有稳健性、适用于高通量筛选，并且能够鉴定出 LH2 的有效特异性抑制剂。
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