Composition Exhibiting a von Willebrand Factor (vWF) Protease Activity Comprising a Polypeptide Chain with the Amino Acid Sequence AAGGILHLELLV

Abstract

The invention relates to vWF cleaving entities having a molecular weight of 180 kD, 170 kD, 160 kD, 120 kD or 110 kD and an N-terminal amino acid sequence of AAGGILHLELLV, vWF cleaving complexes and methods for their production.

What is claimed: 
     
         1 . An pharmaceutical composition comprising an effective dose of a recombinant polypeptide for the treatment of a disease in which the patient has a supranormal vWF content or an increased level of high-molecular weight vWF, wherein the polypeptide exhibits vWF protease activity and vWF binding activity, is produced in an in vitro mammalian cell culture system, and has a molecular weight between about 180 kD and about 120 kD as determined by SDS-PAGE under reducing conditions and comprising the amino acid sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 4) 
                 
                     
                   AAGGILHLELLVAVGPDVFQAHQEDTERYVLTNLNIGAELLRDPS 
                 
                     
                     
                 
                     
                   LGAQFRVHLVKMVILTEPEGAPNITANLTSSLLSVCGWSQTINPE 
                 
                     
                     
                 
                     
                   DDTDPGHADLVLYITRFDLELPDGNRQVRGVTQLGGACSPTWSCL 
                 
                     
                     
                 
                     
                   ITEDTGFDLGVTI. 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the in vitro mammalian cell culture system is a CHO cell culture system. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the disease is thrombotic thrombocytopenic purpura (TTP), Henoch-Schonlein purpura, preeclampsia, neonatal thrombocytopenia, or hemolytic-uremic syndrome. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the recombinant polypeptide comprises the amino acid sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 4) 
                 
                     
                   AAGGILHLELLVAVGPDVFQAHQEDTERYVLTNLNIGAELLRDPS 
                 
                     
                     
                 
                     
                   LGAQFRVHLVKMVILTEPEGAPNITANLTSSLLSVCGWSQTINPE 
                 
                     
                     
                 
                     
                   DDTDPGHADLVLYITRFDLELPDGNRQVRGVTQLGGACSPTWSCL 
                 
                     
                     
                 
                     
                   ITEDTGFDLGVTI. 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . The pharmaceutical composition of  claim 3 , wherein the disease is thrombotic thrombocytopenic purpura (TTP), Henoch-Schonlein purpura, preeclampsia, neonatal thrombocytopenia, or hemolytic-uremic syndrome. 
     
     
         6 . A method of treating a disease in which a patient has a supranormal vWF content or an increased level of high-molecular weight vWF, wherein the method comprises administering a pharmaceutical composition comprising an effective dose of an isolated polypeptide that has vWF protease activity and comprises a peptide chain that has a molecular weight from about 180 kD to about 120 kD as determined by SDS-PAGE under reducing conditions and comprises the amino acid sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 4) 
                 
                     
                   AAGGILHLELLVAVGPDVFQAHQEDTERYVLTNLNIGAELLRDPS 
                 
                     
                     
                 
                     
                   LGAQFRVHLVKMVILTEPEGAPNITANLTSSLLSVCGWSQTINPE 
                 
                     
                     
                 
                     
                   DDTDPGHADLVLYITRFDLELPDGNRQVRGVTQLGGACSPTWSCL 
                 
                     
                     
                 
                     
                   ITEDTGFDLGVTI. 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         7 . The method of  claim 7 , wherein the disease is thrombotic throbocytic purpura (TTP), Henoch-Schonlein purpura, preeclampsia, neonatal thrombocytopenia or hemolyticuremic syndrome.