Small molecule inhibitors of urokinase-type plasminogen activator

The urokinase-type plasminogen activator (uPA) protein is a multifunctional protein involved in a myriad of biological activities including extracellular matrix degradation and cell invasion. Active uPA is a 411 amino acid protein consisting of 3 domains, each of which confers a particular biological function to the overall protein. The amino terminal domain or growth factor domain (GFD), comprised of amino acid residues 1 – 48, is involved in uPA interaction with its cell surface receptor, urokinase-type plasminogen activator receptor (UPAR). The interaction of uPA with UPAR promotes, in part, cell adhesion, migration and invasion. A second domain is the kringle domain, comprising amino acid residues 49 – 135. Initially thought to bind heparin, the kringle domain has more recently been shown to possess antiangiogenic activity. A third domain comprising amino acid residues 159 – 411, the serine protease domain, is involved in the proteolytic activation of plasminogen to plasmin. The production of plasmin by uPA begins a cascade of events manifested by extracellular matrix degradation. The recent patent literature describes small molecule compounds, which inhibit the interaction of uPA with UPAR, inhibit the proteolytic activity of the uPA serine protease domain and inhibit the interaction of uPA with its natural inhibitor, plasminogen activator inhibitor-1 (PAI-1). Small peptides encompassing residues 19 – 31 of the GFD have been developed which exhibit potent inhibition of the uPA–UPAR interaction and show efficacy in tumour-bearing animal models. Small molecules have been disclosed by Corvas, which are reported to be inhibitors of PAI-1. Finally, two approaches toward the development of inhibitors of the uPA serine protease domain have been described in the recent patent literature. The first approach describes non-covalent peptidederived inhibitors discovered by phage display techniques, which bind in the substrate-binding groove of the uPA active site. An alternative approach describes non-covalent small molecule inhibitors, which bind in the enzyme active site in a slightly different binding mode than the peptide-derived inhibitors. These small molecule non-peptide analogues inhibit the uPA proteolytic activity quite effectively and are reported to possess excellent enzyme selectivity and highly improved oral activity. The clinical utility of small molecule uPA enzyme inhibitor analogues awaits the results of a preliminary clinical evaluation of compounds described by Wilex.