Compositions and methods for counteracting factor xa inhibition

Abstract

The disclosure provides compositions and methods for counteracting the effects of direct activated Factor X (FXa) inhibitors in a subject by administering a variant of FXa.

What is claimed is: 
     
         1 . A method for reducing or preventing bleeding in a subject being treated with a direct Factor Xa inhibitor, comprising administering to said subject a Factor Xa variant that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         2 . A method for reducing or preventing bleeding in a subject being treated with rivaroxaban or apixaban, comprising administering to said subject a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         3 . A pharmaceutical composition for reducing or preventing bleeding in a subject being treated with a direct Factor Xa inhibitor, comprising a Factor Xa variant that that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         4 . A pharmaceutical composition for reducing or preventing bleeding in a subject being treated with rivaroxaban or apixaban, comprising a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         5 . Use of a Factor Xa variant in the production of a medicament for reducing or preventing bleeding in a subject being treated with a direct Factor Xa inhibitor, wherein the Factor Xa variant contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         6 . Use of a Factor Xa variant in the production of a medicament for reducing or preventing bleeding in a subject being treated with rivaroxaban or apixaban, wherein the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         7 . The method, composition or use of any one of  claims 1 - 6 , wherein there is a reduction in bleeding of at least about 5%-10%, 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95%, or 95%-100%. 
     
     
         8 . A method for increasing the amount of thrombin produced in the presence of a direct Factor Xa inhibitor in a subject in need thereof, comprising administering to said subject a Factor Xa variant that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         9 . A method for increasing the amount of thrombin produced in the presence of rivaroxaban or apixaban in a subject in need thereof, comprising administering to said subject a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         10 . A pharmaceutical composition for increasing the amount of thrombin produced in the presence of rivaroxaban or apixaban in a subject in need thereof, comprising a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         11 . A pharmaceutical composition for increasing the amount of thrombin produced in the presence of a direct Factor Xa inhibitor in a subject in need thereof, comprising a Factor Xa variant that that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         12 . Use of a Factor Xa variant in the production of a medicament for increasing the amount of thrombin produced in the presence of rivaroxaban or apixaban, wherein the Factor Xa variant contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         13 . Use of a Factor Xa variant in the production of a medicament for increasing the amount of thrombin produced in the presence of a direct Factor Xa inhibitor, wherein the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         14 . The method, composition or use of any one of  claims 8 - 13 , wherein the amount of thrombin produced increases at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 100%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, or 50-fold. 
     
     
         15 . A method for decreasing clotting time in the presence of a direct Factor Xa inhibitor in a subject in need thereof, comprising administering to said subject a Factor Xa variant that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         16 . A method for decreasing clotting time in the presence of rivaroxaban or apixaban in a subject in need thereof, comprising administering to said subject a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         17 . A pharmaceutical composition for decreasing clotting time in the presence of a direct Factor Xa inhibitor in a subject in need thereof, comprising a Factor Xa variant that that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         18 . A pharmaceutical composition for decreasing clotting time in the presence of rivaroxaban or apixaban in a subject in need thereof, comprising a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         19 . Use of a Factor Xa variant in the production of a medicament for decreasing clotting time in the presence of a direct Factor Xa inhibitor in a subject in need thereof, wherein the Factor Xa variant contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         20 . Use of a Factor Xa variant in the production of a medicament for decreasing clotting time in the presence of rivaroxaban or apixaban in a subject in need thereof, wherein the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         21 . The method, composition or use of any one of  claims 15 - 20 , wherein there is a reduction in clotting time of at least about 5%-10%, 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95%, or 95%-100%. 
     
     
         22 . The method, composition or use of any one of  claims 15 - 20 , wherein the reduction in clotting time is measured using prothrombin time (PT). 
     
     
         23 . The method of  claim 22 , wherein said PT in said subject is about 25 seconds, 24 seconds, 23 seconds, 22 seconds, 21 seconds, 20 seconds, 19 seconds, 18 seconds, 17 seconds, 16 seconds, 15 seconds, 14 seconds, 13 seconds, 12 seconds, 11 seconds, or 10 seconds. 
     
     
         24 . The method of  claim 22 , wherein the International Normalized Ratio (INR) in said subject is about 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, or 0.7. 
     
     
         25 . The method of any one of  claim 22 ,  23 , or  24 , wherein PT is determined 15 mins, 20 mins, 30 mins, 40 mins, 45 mins, 50 mins, 60 mins, 75 min, or 90 min after administration of the FXa variant. 
     
     
         26 . A pharmaceutical composition comprising a Factor Xa variant that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
       wherein said Factor Xa variant counters the effect of a direct Factor Xa inhibitor at a plasma concentration of at least 100-fold lower than the plasma concentration of the direct Factor Xa inhibitor. 
     
     
         27 . A pharmaceutical composition comprising a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr, and wherein the Factor Xa variant counters the effect of rivaroxaban or apixaban at a plasma concentration of at least 100-fold lower than the plasma concentration of the rivaroxaban or apixaban. 
     
     
         28 . The method, composition or use of any of the previous claims, wherein the Factor Xa variant counters the effect of a direct Factor Xa inhibitor at a plasma concentration of at least 100-fold lower than the plasma concentration of the direct Factor Xa inhibitor. 
     
     
         29 . The method, composition or use of any of the previous claims, wherein the Factor Xa variant is administered before a planned surgery, after an injury or after a direct Factor Xa inhibitor overdose. 
     
     
         30 . The method, composition or use of any of the previous claims, wherein Factor Xa variant is administered more than one time. 
     
     
         31 . The method, composition or use of any of the previous claims, wherein at least one additional procoagulant is administered. 
     
     
         32 . The method, composition or use of  claim 30 , wherein the procoagulant is selected from the group consisting of: a different Factor Xa variant, Factor IX, Factor XIa, Factor XIIa, Factor VIII, Factor VIIa, FEIBA and prothrombin complex concentrate (PCC). 
     
     
         33 . The method, composition or use of any of the previous claims, wherein the plasma concentration of the direct FXa inhibitor is a supratherapeutic amount. 
     
     
         34 . The method, composition or use of any of the previous claims, wherein the direct FXa inhibitor is rivaroxaban and wherein the plasma concentration of rivaroxaban is at least about 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, or 800 nM. 
     
     
         35 . The method, composition or use of any of the previous claims, wherein the direct FXa inhibitor is apixaban and wherein the plasma concentration of apixaban is at least about 50 nM, 100 nM, 150 nM, 200nM, 250 nM, 300 nM, 350 nM, or 400 nM. 
     
     
         36 . A method for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, comprising administering to said subject a Factor Xa variant that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         37 . A method for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, comprising administering to said subject a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         38 . A pharmaceutical composition for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, comprising a Factor Xa variant that that contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         39 . A pharmaceutical composition for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, comprising a Factor Xa variant in which the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr. 
     
     
         40 . Use of a Factor Xa variant in the production of a medicament for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, wherein the Factor Xa variant contains at least one modification selected from the group consisting of:
 a) the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Thr, Leu, Phe, Asp or Gly; and   b) the amino acid at the position corresponding to 236 in SEQ ID NO:1 is substituted with Leu, Ala, or Gly.   
     
     
         41 . Use of a Factor Xa variant in the production of a medicament for effecting the urgent reversal of acquired coagulopathy due to FXa inhibition therapy in a subject with acute major bleeding, wherein the amino acid at the position corresponding to 235 in SEQ ID NO:1 is substituted with Leu or Thr.