Development of a Rapid Fluorescence-Based High-Throughput Screening Assay to Identify Novel Kynurenine 3-Monooxygenase Inhibitor Scaffolds

被引：10
作者：
Jacobs, K. R. [1 ]
Guillemin, G. J. [1 ]
Lovejoy, D. B. [1 ]
机构：
[1] Macquarie Univ, Fac Med & Hlth Sci, Dept Biomed Sci, Neuroinflammat Grp,Ctr Motor Neuron Dis Res, Sydney, NSW, Australia
来源：
SLAS DISCOVERY | 2018年 / 23卷 / 06期
关键词：
kynurenine; 3-monooxygenase; high-throughput screening; 3-hydroxykynurenine; neurodegeneration; NADPH fluorescence; HUNTINGTONS-DISEASE; QUINOLINATE LEVELS; PATHWAY; 3-HYDROXYKYNURENINE; ENZYME; BRAIN; ACID; MICE;
D O I：
10.1177/2472555218757180
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Kynurenine 3-monooxygenase (KMO) is a well-validated therapeutic target for the treatment of neurodegenerative diseases, including Alzheimer's disease (AD) and Huntington's disease (HD). This work reports a facile fluorescence-based KMO assay optimized for high-throughput screening (HTS) that achieves a throughput approximately 20-fold higher than the fastest KMO assay currently reported. The screen was run with excellent performance (average Z' value of 0.80) from 110,000 compounds across 341 plates and exceeded all statistical parameters used to describe a robust HTS assay. A subset of molecules was selected for validation by ultra-high-performance liquid chromatography, resulting in the confirmation of a novel hit with an IC50 comparable to that of the well-described KMO inhibitor Ro-61-8048. A medicinal chemistry program is currently underway to further develop our novel KMO inhibitor scaffolds.
引用
收藏
页码：554 / 560
页数：7
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