Anticoagulation agent and uses thereof

Abstract

The present invention provides an anticoagulant agent including a first element capable of inhibiting coagulation and a second element capable of targeting an activated platelet wherein upon administration of the agent to a subject the second element directs the first element to the activated platelet. Also provided is a probe for detecting a blood vessel abnormality including (a) a binding element capable of targeting an activated platelet and (b) a label. Applicant has shown that agents and probes directed to activated platelets are useful in the diagnosis and therapy of coagulation disorders.

1 - 50 . (canceled)  
   
   
       51 . A composition comprising: 
 a) a single chain antibody or derivative thereof, wherein, 
 i) the single chain antibody or derivative thereof specifically binds the activated state of the platelet integrin receptor GP IIb/IIIa; and  
 ii) the single chain antibody or derivative thereof has substantially no effect upon thrombosis when bound to the activated state of the platelet integrin receptor GP IIb/IIIa.  
   
   
   
       52 . The composition of  claim 51 , further comprising a label bound to the single chain antibody or derivative thereof.  
   
   
       53 . The composition of  claim 52 , wherein the label comprises a contrast agent for magnetic resonance imaging.  
   
   
       54 . The composition of  claim 52 , wherein the label comprises a paramagnetic bead.  
   
   
       55 . The composition of  claim 52 , wherein a histidine-tag is bound to the antibody or derivative thereof.  
   
   
       56 . The composition of  claim 55 , wherein a label is conjugated to the histidine-tag.  
   
   
       57 . The composition of  claim 56 , wherein the label comprises a paramagnetic bead.  
   
   
       58 . The composition of  claim 56 , wherein the label comprises a superparamagnetic iron oxide particle (SPIO).  
   
   
       59 . The composition of  claim 56 , wherein the label comprises a micron-sized paramagnetic iron oxide (MPIO).  
   
   
       60 . The composition of  claim 59 , wherein the label comprises an autofluorescent cobalt-functionalized MPIO.  
   
   
       61 . The composition of  claim 51 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a heavy variable chain, the heavy variable chain comprising an amino acid sequence selected from the group consisting of GFTFSSYIMS (SEQ ID NO: 7), TIRSGGDNTYYPDSVKG (SEQ ID NO: 8), and YYGNYGGLAY (SEQ ID NO: 9).  
   
   
       62 . The composition of  claim 51 , wherein the heavy variable chain comprises the amino acid sequence beginning at position 36 and ending at position 158 of SEQ ID NO: 2.  
   
   
       63 . The composition of  claim 51 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a light variable chain, the light variable chain comprising an amino acid sequence selected from the group consisting of RASGNIHNYLA (SEQ ID NO: 10), NAKTLAD (SEQ ID NO: 11), and QHFWSTPYT (SEQ ID NO: 12).  
   
   
       64 . The composition of  claim 51 , wherein the light variable chain comprises the amino acid sequence beginning at position 168 and ending at position 284 of SEQ ID NO: 2.  
   
   
       65 . A method of assaying for an activated blood platelet comprising: 
 a) contacting blood or tissue with a single chain antibody or derivative thereof, wherein, 
 i) the single chain antibody or derivative thereof specifically binds the activated state of the platelet integrin receptor GP IIb/IIIa; and  
 ii) the single chain antibody or derivative thereof has substantially no effect upon thrombosis when bound to the activated state of the platelet integrin receptor GP IIb/IIIa  
   under conditions that provide for specific binding of the antibody or derivative thereof with the activated state of the platelet integrin receptor GP IIb/IIIa to form a bound complex; and    b) imaging the bound complex, wherein an image of the bound complex signals the detection of an activated blood platelet.    
   
   
       66 . The method of  claim 65 , wherein the bound complex is imaged by a second antibody specific to the single chain antibody or derivative thereof.  
   
   
       67 . The method of  claim 65 , wherein the single chain antibody or derivative thereof is coupled to a label.  
   
   
       68 . The method of  claim 67 , wherein the label comprises a contrast agent for magnetic resonance imaging.  
   
   
       69 . The method of  claim 67 , wherein the label comprises a paramagnetic bead.  
   
   
       70 . The method of  claim 67 , wherein a histidine-tag is bound to the antibody or derivative thereof.  
   
   
       71 . The method of  claim 70 , wherein the bound complex is imaged by detection of the histidine-tag.  
   
   
       72 . The method of  claim 70 , wherein a label is conjugated to the histidine-tag.  
   
   
       73 . The method of  claim 72 , wherein the label comprises a paramagnetic bead.  
   
   
       74 . The method of  claim 73 , wherein the label comprises a superparamagnetic iron oxide particle (SPIO).  
   
   
       75 . The method of  claim 73 , wherein the label comprises a micron-sized paramagnetic iron oxide (MPIO).  
   
   
       76 . The method of  claim 75 , wherein the label comprises an autofluorescent cobalt-functionalized MPIO.  
   
   
       77 . The method of  claim 65 , wherein the imaging comprises magnetic resonance imaging (MRI).  
   
   
       78 . The method of  claim 65 , wherein the platelet integrin receptor GP IIb/IIIa is activated by the binding of fibrinogen or its mimetics.  
   
   
       79 . The method of  claim 65 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a heavy variable chain, the heavy variable chain comprising an amino acid sequence selected from the group consisting of GFTFSSYIMS (SEQ ID NO: 7), TIRSGGDNTYYPDSVKG (SEQ ID NO: 8), and YYGNYGGLAY (SEQ ID NO: 9).  
   
   
       80 . The method of  claim 65 , wherein the heavy variable chain comprises the amino acid sequence beginning at position 36 and ending at position 158 of SEQ ID NO: 2.  
   
   
       81 . The method of  claim 65 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a light variable chain, the light variable chain comprising an amino acid sequence selected from the group consisting of RASGNIHNYLA (SEQ ID NO: 10), NAKTLAD (SEQ ID NO: 11), and QHFWSTPYT (SEQ ID NO: 12).  
   
   
       82 . The method of  claim 65 , wherein the light variable chain comprises the amino acid sequence beginning at position 168 and ending at position 284 of SEQ ID NO: 2.  
   
   
       83 . The method of  claim 65 , wherein the activated platelet is in vivo.  
   
   
       84 . The method of  claim 65 , wherein the activated platelet is ex vivo.  
   
   
       85 . A therapeutic composition comprising: 
 a) a single chain antibody or derivative thereof, wherein, 
 i) the single chain antibody or derivative thereof specifically binds the activated state of the platelet integrin receptor GP IIb/IIIa; and  
 ii) the single chain antibody or derivative thereof has substantially no effect upon thrombosis when bound to the activated state of the platelet integrin receptor GP IIb/IIIa; and  
   b) a pharmaceutically active element bound to the single chain antibody or derivative thereof.    
   
   
       86 . The composition of  claim 85 , wherein the pharmaceutically active element comprises an anticoagulant.  
   
   
       87 . The composition of  claim 86 , wherein the pharmaceutically active element is tick anticoagulant protein (TAP).  
   
   
       88 . The composition of  claim 86 , wherein the pharmaceutically active element is hirudin or a hirudin derivative.  
   
   
       89 . The composition of  claim 85 , wherein a histidine-tag is bound to the antibody or derivative thereof.  
   
   
       90 . The composition of  claim 89 , wherein the pharmaceutically active element is bound to the histidine-tag.  
   
   
       91 . The composition of  claim 88 , wherein the pharmaceutically active element is tick anticoagulant protein (TAP).  
   
   
       92 . The composition of  claim 88 , wherein the pharmaceutically active element is hirudin or a hirudin derivative.  
   
   
       93 . The composition of  claim 85 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a heavy variable chain, the heavy variable chain comprising an amino acid sequence selected from the group consisting of GFTFSSYIMS (SEQ ID NO: 7), TIRSGGDNTYYPDSVKG (SEQ ID NO: 8), and YYGNYGGLAY (SEQ ID NO: 9).  
   
   
       94 . The composition of  claim 85 , wherein the heavy variable chain comprises the amino acid sequence beginning at position 36 and ending at position 158 of SEQ ID NO: 2.  
   
   
       95 . The composition of  claim 85 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a light variable chain, the light variable chain comprising an amino acid sequence selected from the group consisting of RASGNIHNYLA (SEQ ID NO: 10), NAKTLAD (SEQ ID NO: 11), and QHFWSTPYT (SEQ ID NO: 12).  
   
   
       96 . The composition of  claim 85 , wherein the light variable chain comprises the amino acid sequence beginning at position 168 and ending at position 284 of SEQ ID NO: 2.  
   
   
       97 . A method of treating a coagulation disorder comprising administering a single chain antibody or derivative thereof, wherein, 
 i) the single chain antibody or derivative thereof specifically binds the activated state of the platelet integrin receptor GP IIb/IIIa; and    ii) the single chain antibody or derivative thereof has substantially no effect upon thrombosis when bound to the activated state of the platelet integrin receptor GP IIb/IIIa; and    b) a pharmaceutically active element bound to the single chain antibody or derivative thereof.    
   
   
       98 . The method of  claim 97 , wherein the pharmaceutically active element comprises an anticoagulant.  
   
   
       99 . The method of  claim 98 , wherein the pharmaceutically active element is tick anticoagulant protein (TAP).  
   
   
       100 . The method of  claim 99 , wherein the pharmaceutically active element is hirudin or a hirudin derivative.  
   
   
       101 . The method of  claim 97 , wherein a histidine-tag is bound to the antibody or derivative thereof.  
   
   
       102 . The method of  claim 101 , wherein the pharmaceutically active element is bound to the histidine-tag.  
   
   
       103 . The method of  claim 102 , wherein the pharmaceutically active element is tick anticoagulant protein (TAP).  
   
   
       104 . The method of  claim 103 , wherein the pharmaceutically active element is hirudin or a hirudin derivative.  
   
   
       105 . The method of  claim 97 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a heavy variable chain, the heavy variable chain comprising an amino acid sequence selected from the group consisting of GFTFSSYIMS (SEQ ID NO: 7), TIRSGGDNTYYPDSVKG (SEQ ID NO: 8), and YYGNYGGLAY (SEQ ID NO: 9).  
   
   
       106 . The method of  claim 97 , wherein the heavy variable chain comprises the amino acid sequence beginning at position 36 and ending at position 158 of SEQ ID NO: 2.  
   
   
       107 . The method of  claim 97 , wherein the single chain antibody or derivative thereof comprises a segment of an immunoglobulin comprising a light variable chain, the light variable chain comprising an amino acid sequence selected from the group consisting of RASGNIHNYLA (SEQ ID NO: 10), NAKTLAD (SEQ ID NO: 11), and QHFWSTPYT (SEQ ID NO: 12).  
   
   
       108 . The method of  claim 97 , wherein the light variable chain comprises the amino acid sequence beginning at position 168 and ending at position 284 of SEQ ID NO: 2.  
   
   
       109 . An isolated nucleic acid sequence encoding a single chain antibody or derivative thereof, wherein, the single chain antibody or derivative thereof specifically binds the activated state of the platelet integrin receptor GP IIb/IIa; and the single chain antibody or derivative thereof has substantially no effect upon thrombosis when bound to the activated state of the platelet integrin receptor GP IIb/IIIa.  
   
   
       110 . An isolated nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 2.  
   
   
       111 . An isolated and purified polypeptide comprising the sequence of SEQ ID NO: 2.