A Novel Glycomimetic Compound (GMI-1757) with Dual Functional Antagonism to E-Selectin and Galectin-3 Demonstrates Inhibition of Thrombus Formation in an Inferior Vena Cava Model

▪ E-selectin functions in venous thrombosis by binding and activating host cells to initiate the coagulation cascade. The E-selectin antagonist, Uproleselan (GMI-1271), has been shown to attenuate thrombus formation following electrical stimulation in a preclinical inferior vena cava (IVC) model without significantly affecting hemostasis (Culmer DL et al. Thromb Haemost. 117:1171-1181, 2017). A recent study has reported that galectin-3 (gal-3) and gal-3 binding protein are associated with murine thrombogenesis where they interact at the thrombus-vein wall interface, and that gal-3 may be contributing to thrombosis through proinflammatory mechanisms (Elise P et al. Blood 125:1813-1821, 2015). Collectively these studies suggest that pharmacologic targeting of both E-selectin and gal-3 function may afford a new class of therapeutics for treatment of venous thrombosis. In the present studies, we report on the activity of a novel glycomimetic compound with dual functional antagonism of E-selectin and gal-3, and demonstrate its anti-thrombotic activity in an IVC model. Binding affinities of GMI-1757 in solution or to immobilized E-selectin or gal-3 were determined by surface plasmon resonance (SPR) or microscale thermophoresis (MST), respectively. By SPR the mean (+/- SD) KD of GMI-1757 was 61 +/- 4 nM and 276 +/- 98 nM for E-selectin and gal-3, respectively (N=2 determinations). By MST the mean KD of GMI-1757 was 563 +/- 158 nM and 83 +/- 9 nM for E-selectin and gal-3, respectively (N=2-3 determinations). GMI-1757 was subsequently evaluated for its ability to inhibit binding of (a) sialyl Lea to immobilized human E-selectin and (b) gal-3 to an immobilized Galb1-3GlcNAc carbohydrate structure. The median IC50 of GMI-1757 for ligand binding of E-selectin or gal-3 was 800 nM (n=9 independent assays) and 110 nM (n=7 independent assays), respectively. The pharmacokinetics of GMI-1757 was evaluated following intravenous (IV) administration in male CD-1 mice at 5 mg/kg. Blood samples were collected up to 24 hours post dose, and GMI-1757 concentrations were determined with a qualified LC-MS/MS method. Following IV dosing of GMI 1757 at 5 mg/kg, the average half-life was 3.5 hours. Its average clearance rate was 1.39 +/- 0.180 L/hr/kg and the average volume of distribution was 1.35 +/- 0.651 L/kg. The in vivo activity of GMI-1757 following intraperitoneal (IP) administration was assessed in an acute model of IVC thrombosis.Immediately following the induction of a non-oc