Fluorescent diphenylphosphonate-based probes for detection of serine protease activity during inflammation

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS

卷:27

Article

Edgington-Mitchell, Laura E.¹  Barlow, Nicholas²  Aurelio, Luigi²  Samha, Aminath¹  Szabo, Monika²  Graham, Bim²  Bunnett, Nigel^1,3 

[1] Monash Univ, Drug Discovery Biol, Parkville, Vic, Australia

[2] Monash Univ, Monash Inst Pharmaceut Sci, Med Chem, Parkville, Vic, Australia

[3] Univ Melbourne, Dept Pharmacol & Therapeut, Parkville, Vic, Australia

关键词: Fluorescent probes;    Activity-based probes;    Trypsin;    Elastase;    Protease;    Pancreatitis;    Colitis;    Inflammation;    Protease inhibitors;    Diphenylphosphonate;   

DOI  :  10.1016/j.bmcl.2016.11.064

来源: Elsevier

【 摘 要 】

Activity-based probes are small molecules that covalently bind to the active site of a protease in an activity -dependent manner. We synthesized and characterized two fluorescent activity-based probes that target serine proteases with trypsin-like or elastase-like activity. We assessed the selectivity and potency of these probes against recombinant enzymes and demonstrated that while they are efficacious at labeling active proteases in complex protein mixtures in vitro, they are less valuable for in vivo studies. We used these probes to evaluate serine protease activity in two mouse models of acute inflammation, including pancreatitis and colitis. As anticipated, the activity of trypsin-like proteases was increased during pancreatitis. Levels of elastase-like proteases were low in pancreatic lysates and colonic luminal fluids, whether healthy or inflamed. Exogenously added recombinant neutrophil elastase was inhibited upon incubation with these samples, an effect that was augmented in inflamed samples compared to controls. These data suggest that endogenous inhibitors and elastase-degrading proteases are upregulated during inflammation. (C) 2016 Elsevier Ltd. All rights reserved.

【 授权许可】

Free