Background
Human factor XIa (FXIa) is an actively pursued target for development of safer anticoagulants. Our longâstanding hypothesis has been that allosterism originating from heparinâbinding site(s) on coagulation enzymes is a promising approach to yield safer agents.
Objectives
To develop a synthetic heparin mimetic as an inhibitor of FXIa so as to reduce clot formation in vivo but not carry high bleeding risk.
Methods
We employed a gamut of methods involving synthetic chemistry, biophysical biochemistry, enzyme assays, blood and plasma coagulation assays, and in vivo thrombosis models in this work.
Results
Sulfated chiroâinositol (SCI), a nonâsaccharide mimetic of heparin, was synthesized in three steps in overall yields of >50%. SCI inhibited FXIa with potency of 280Â nmol/L and preferentially engaged FXIa's heparinâbinding site to conformationally alter its active site. SCI inhibition of FXIa could be rapidly reversed by common antidotes, such as protamine. SCI preferentially prolonged plasma clotting initiated with recalcification, rather than thromboplastin, alluding to its intrinsic pathwayâbased mechanism. Human blood thromboelastography indicated good ex vivo anticoagulation properties of SCI. Rat tail bleeding and maximumâdoseâtolerated studies indicated that no major bleeding or toxicity concerns for SCI suggesting a potentially safer anticoagulation outcome. FeCl3âinduced arterial and thromboplastinâinduced venous thrombosis model studies in the rat showed reduced thrombus formation by SCI at 250Â ÎŒg/animal, which matched enoxaparin at 2500Â ÎŒg/animal.
Conclusions
Overall, SCI is a highly promising, allosteric inhibitor of FXIa that induces potent anticoagulation in vivo. Further studies are necessary to assess SCI in animal models mimicking human clinical indications.