Lyophilized formulations for factor xa antidote

Abstract

The present disclosure relates to solutions and methods of preparing lyophilized formulations of factor Xa (fXa) antidotes. A suitable aqueous formulation suitable for lyophilization can include a fXa antidote, a solubilizing agent, and a stabilizer, wherein the formulation does not collapse during lyophilization.

1 . A lyophilized composition obtainable by lyophilizing an aqueous formulation, wherein:
 the aqueous formulation comprises a stabilizer, from 25 mM to 110 mM arginine, and at least 15 mg/mL of a two-chain polypeptide comprising a first chain comprising the amino acid sequence of SEQ ID NO. 4, and a second chain comprising the amino acid sequence of SEQ ID NO. 5, wherein the polypeptide cannot assemble into a prothrombinase complex;   the aqueous formulation has a pH from 7.5 to 8;   the stabilizer comprises from 2% to 7% sucrose (w/v) and from 0% to 5% mannitol (w/v); and   the molar ratio of the stabilizer to the polypeptide is at least 100.   
     
     
         2 . The composition of  claim 1 , wherein the aqueous formulation comprises from 40 mM to 50 mM arginine, from 5.5% to 7.5% sucrose (w/v), and 20 mg/mL of the polypeptide. 
     
     
         3 . The composition of  claim 1 , wherein the aqueous formulation comprises from 90 mM to 110 mM arginine, from 5.5% to 7.5% sucrose (w/v), and at least 30 mg/mL of the polypeptide. 
     
     
         4 . The composition of  claim 3 , wherein the aqueous formulation comprises 45 mg/mL of the polypeptide. 
     
     
         5 . The composition of any of the above claims, wherein the aqueous formulation comprises no mannitol. 
     
     
         6 . The composition of  claim 1 , wherein the aqueous formulation comprises no mannitol and 40 mg/mL of the polypeptide. 
     
     
         7 . The composition of any of the above claims, wherein the polypeptide comprises an amino acid residue that is modified to be different from natural amino acids. 
     
     
         8 . The composition of  claim 6 , wherein residue Asp29 of the first chain is modified to (3R)-3-hydroxyAsp at Asp29. 
     
     
         9 . The composition of any of  claims 1 - 6 , wherein the polypeptide comprises at least an intra-chain disulfide bond for each of the first and second chains. 
     
     
         10 . A lyophilized composition obtainable by lyophilizing an aqueous formulation, wherein:
 the aqueous formulation comprises about 45 mM arginine, about 6% sucrose (w/v), 0% mannitol, and about 20 mg/mL of a two-chain polypeptide comprising a first chain comprising the amino acid sequence of SEQ ID NO. 4, and a second chain comprising the amino acid sequence of SEQ ID NO. 5, wherein the polypeptide cannot assemble into a prothrombinase complex, and   the aqueous formulation has a pH of about 7.8.   
     
     
         11 . A lyophilized composition obtainable by lyophilizing an aqueous formulation, wherein:
 the aqueous formulation comprises about 100 mM arginine, about 6% sucrose (w/v), 0% mannitol, and about 40 mg/mL of a two-chain polypeptide comprising a first chain comprising the amino acid sequence of SEQ ID NO. 4, and a second chain comprising the amino acid sequence of SEQ ID NO. 5, wherein the polypeptide cannot assemble into a prothrombinase complex; and   the aqueous formulation has a pH of about 7.8.   
     
     
         12 . The composition of  claim 11 , wherein the two-chain polypeptide has modifications to the Gla domain and the active site as compared to the wild-type fXa protein, is able to bind to a fXa inhibitor but does not assemble into a prothrombinase complex. 
     
     
         13 . The composition of  claim 1 , wherein the glass transition temperature is from about −27° C. to about −37° C. 
     
     
         14 . The composition of any of the above claims, wherein the aqueous formulation further comprises a surfactant and a buffer. 
     
     
         15 . A method of preparing the lyophilized, amorphous composition of any one of  claims 1 - 14 . 
     
     
         16 . A method of reducing bleeding in a subject undergoing anticoagulant therapy with a factor Xa inhibitor comprising administering to the subject a solution prepared by dissolving the composition of any one of  claims 1 - 14  in an aqueous solvent. 
     
     
         17 . The method of  claim 16 , wherein the factor Xa inhibitor is apixaban, rivaroxaban or betrixaban.