Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents.

Abstract

Thrombosis, or blood clot formation, and its sequelae remain a leading cause of morbidity and mortality, and recurrent thrombosis is common despite current optimal therapy. Protein disulfide isomerase (PDI) is an oxidoreductase that has recently been shown to participate in thrombus formation. While currently available antithrombotic agents inhibit either platelet aggregation or fibrin generation, inhibition of secreted PDI blocks the earliest stages of thrombus formation, suppressing both pathways. Here, we explored extracellular PDI as an alternative target of antithrombotic therapy. A high-throughput screen identified quercetin-3-rutinoside as an inhibitor of PDI reductase activity in vitro. Inhibition of PDI was selective, as quercetin-3-rutinoside failed to inhibit the reductase activity of several other thiol isomerases found in the vasculature. Cellular assays showed that quercetin-3-rutinoside inhibited aggregation of human and mouse platelets and endothelial cell-mediated fibrin generation in human endothelial cells. Using intravital microscopy in mice, we demonstrated that quercetin-3-rutinoside blocks thrombus formation in vivo by inhibiting PDI. Infusion of recombinant PDI reversed the antithrombotic effect of quercetin-3-rutinoside. Thus, PDI is a viable target for small molecule inhibition of thrombus formation, and its inhibition may prove to be a useful adjunct in refractory thrombotic diseases that are not controlled with conventional antithrombotic agents.

• Authors: Reema Jasuja, Freda H Passam, Daniel R Kennedy, Sarah H Kim, Lotte van Hessem, Lin Lin, Sheryl R Bowley, Sucharit S Joshi, James R Dilks, Bruce Furie, Barbara C Furie, Robert Flaumenhaft
• Journal: The Journal of clinical investigation (J Clin Invest) Vol. 122 Issue 6 Pg. 2104-13 (Jun 2012) ISSN: 1558-8238 [Electronic] United States
• PMID: 22565308 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Enzyme Inhibitors
• Fibrinolytic Agents
• Recombinant Proteins
• Rutin
• Fibrin
• Protein Disulfide-Isomerases

Topics

• Animals
• Blood Platelets (metabolism)
• Enzyme Inhibitors (pharmacology)
• Fibrin (genetics, metabolism)
• Fibrinolytic Agents (pharmacology)
• Humans
• Mice
• Platelet Aggregation (drug effects)
• Protein Disulfide-Isomerases (adverse effects, antagonists & inhibitors, genetics, pharmacology)
• Recombinant Proteins (adverse effects, antagonists & inhibitors, genetics, pharmacology)
• Rutin (pharmacology)
• Thrombosis (chemically induced, drug therapy, enzymology)