Non-peptide-based fluorogenic small-molecule probe for elastase.
Human neutrophil elastase (HNE) has been identified as a potential therapeutic target for the discovery of anti-inflammatory drugs for decades. However, little progress has been made on assays measuring the activity of HNE, especially on synthetic substrates which play essential role in determination of HNE activity. Herein, a small-molecule compound, 2,2,3,3,3-pentafluoro-N-(2-oxo-4-(trifluoromethyl)-2H-chromen-7-yl)-propanamide (compound 4), has been successfully designed as the first ever non-peptide-based fluorogenic substrate for HNE. A "turn-on" fluorometric assay based on 4 has been successfully developed for rapid determination of HNE activity and the inhibitory kinetic study. Most importantly, the probe 4 shows highly specific response for HNE among seven tested hydrolases or proteins and can be directly used to detect the elevated HNE activity in the serum of chronic obstructive pulmonary disease (COPD) patients compared to that of healthy controls. This specific and cost-effective probe will facilitate future high-throughput discovery of HNE inhibitors and clinical diagnosis of elastase-related diseases.
Key Laboratory of Pesticide & Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University , Wuhan 430079, People's Republic of China.
Qi
Q
Sun
Jun
J
Li
Wan-Nian
WN
Liu
Qing-Jian
QJ
Dong
Wen-Chao
WC
Yang
Guang-Fu
GF
Yang
D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D010196:Pancreatic Elastase / Q000032:analysis*; D014961:X-Ray Diffraction / Q000379:methods
D005456:Fluorescent Dyes; D010196:Pancreatic Elastase