Single domain binding molecule

Abstract

The present invention provides a single domain specific binding molecule having the structure FW1-CDR1-FW2-HV2-FW3a-HV4-FW3b-CDR3-FW4 in which the Framework Regions FW1, FW2, FW3a, FW3b, and FW4, the Complementarity Determining Regions CDR1 and CDR3, and the Hypervariable Regions HV2, and HV4 have amino acid sequences as defined which provide a high affinity anti-human serum albumin (HSA) binding domain.

1 - 16 . (canceled) 
     
     
         17 . A single domain specific binding molecule comprising the structure
   FW1-CDR1-FW2-HV2-FW3a-HV4-FW3b-CDR3-FW4   in which the Framework Regions FW1, FW2, FW3a, FW3b, and FW4, the Complementarity Determining Regions CDR1 and CDR3, and the Hypervariable Regions HV2, and HV4 comprise amino acid sequences in which   FW1 comprises a sequence at least 50% identical to the sequence TRVDQTPRTATRETGESLTINCVLT;   FW2 comprises a sequence at least 50% identical to the sequence TYWYRKNPGS;   FW3a comprises a sequence at least 50% identical to the sequence GRYVESVN;   FW3b comprises a sequence at least 50% identical to the sequence FSLRIKDLTVADSATYICRA;   FW4 comprises a sequence at least 50% identical to the sequence GAGTVLTVN;   CDR1 comprises a sequence at least 50% identical to the sequence DTSYPLYS;   CDR3 comprises a sequence at least 50% identical to a sequence selected from   
       
         
           
                 
                 
               
                   (i) 
                   the sequence MGTNIWTGD, 
                 
                     
                 
                   (ii) 
                   a sequence selected from MATNIWTGD, 
                 
                     
                   MGTDSWTGD, MGTNSWTGD, MSTNIWTGD, 
                 
                     
                   ITTDSWTSD, MGANSWTGD, MGTNGWTGD, SDIAMGTYD, 
                 
                     
                   ITTHSWSGD, LSTYMEAGD, and MDTSAGVVD, 
                 
                     
                 
                   (iii) 
                   a sequence selected from 
                 
                     
                   ESPPICTSQGIAAVTKYYD, YTIHIKLEGH, 
                 
                     
                   HAGYGVWNRGLQWRGYDGYD, YTPGREDY, 
                 
                     
                   EKGRKGSAITSCRRSSYYD, and QSLAISTRSYWYD, 
                 
                     
                   or 
                 
                     
                 
                   (iv) 
                   the sequence GVAGGYCDYALCSSRYAE; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         HV2 comprises a sequence at least 50% identical to the sequence SNKEQISIS; and 
         HV4 comprises a sequence at least 50% identical to the sequence KGTKS. 
       
     
     
         18 . A single domain specific binding molecule as claimed in  claim 17  comprising the structure
   FW1-CDR1-FW2-HV2-FW3a-HV4-FW3b-CDR3-FW4 
 in which the Framework Regions FW1, FW2, FW3a, FW3b, and FW4, the Complementarity Determining Regions CDR1 and CDR3, and the Hypervariable Regions HV2, and HV4 comprise amino acid sequences in which 
 FW1 comprises the sequence TRVDQTPRTATRETGESLTINCVLT; 
 FW2 comprises the sequence TYWYRKNPGS; 
 FW3a comprises the sequence GRYVESVN; 
 FW3b comprises the sequence FSLRIKDLTVADSATYICRA; 
 FW4 comprises the sequence GAGTVLTVN; 
 CDR1 comprises the sequence DTSYPLYS; 
 CDR3 comprises a sequence selected from 
 
       
         
           
                 
                 
               
                   (i) 
                   the sequence MGTNIWTGD, 
                 
                     
                 
                   (ii) 
                   a sequence selected from MATNIWTGD, 
                 
                     
                   MGTDSWTGD, MGTNSWTGD, MSTNIWTGD, 
                 
                     
                   ITTDSWTSD, MGANSWTGD, MGTNGWTGD, SDIAMGTYD, 
                 
                     
                   ITTHSWSGD, LSTYMEAGD, and MDTSAGVVD, 
                 
                     
                 
                   (iii) 
                   a sequence selected from 
                 
                     
                   ESPPICTSQGIAAVTKYYD, YTIHIKLEGH, 
                 
                     
                   HAGYGVWNRGLQWRGYDGYD, YTPGREDY, 
                 
                     
                   EKGRKGSAITSCRRSSYYD, and QSLAISTRSYWYD, 
                 
                     
                   or 
                 
                     
                 
                   (iv) 
                   the sequence GVAGGYCDYALCSSRYAE; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         HV2 comprises the sequence SNKEQISIS; and 
         HV4 comprises the sequence KGTKS. 
       
     
     
         19 . The single domain specific binding molecule as claimed in  claim 17 , which is humanized. 
     
     
         20 . A single domain specific binding molecule comprising an amino acid sequence at least 50% identical to an amino acid sequence selected from E06, BB11 and H08, as shown in  FIG. 1 . 
     
     
         21 . A single domain specific binding molecule as claimed in  claim 20 , comprising an amino acid sequence selected from E06, BB11 and H08, as shown in  FIG. 1 . 
     
     
         22 . A single domain specific binding molecule comprising an amino acid sequence at least 50% identical to an amino acid sequence selected from: huE06 v1.1, huH08 v1.1, huE06 v1.2, huE06 v1.3, huE06 v1.4, huE06 v1.5, huE06 v1.6, huE06 v1.7, huE06 v1.8, huE06 v1.9, huE06 v1.10, AC9, AD4, AG11, AH7, BA11, BB10, BC3, BD12, BE4, and BH4, as shown in  FIG. 2 . 
     
     
         23 . The single domain specific binding molecule as claimed in  claim 22 , comprising an amino acid sequence selected from: huE06 v1.1, huH08 v1.1, huE06 v1.2, huE06 v1.3, huE06 v1.4, huE06 v1.5, huE06 v1.6, huE06 v1.7, huE06 v1.8, huE06 v1.9, huE06 v1.10, AC9, AD4, AG11, AH7, BA11, BB10, BC3, BD12, BE4, and BH4, as shown in  FIG. 2 . 
     
     
         24 . A fusion protein comprising a single domain specific binding molecule as claimed in  claim 17 . 
     
     
         25 . The fusion protein as claimed in  claim 24 , in which the single domain specific binding molecule is fused to a biologically active protein. 
     
     
         26 . A nucleic acid encoding a single domain specific binding molecule as claimed in  claim 17 . 
     
     
         27 . A nucleic acid construct comprising the nucleic acid as claimed in  claim 26 . 
     
     
         28 . A host cell comprising the nucleic acid construct as claimed in  claim 27 . 
     
     
         29 . A process for the production of a single domain specific binding molecule, comprising culturing the host cell as claimed in  claim 28  under conditions suitable for production of the single domain specific binding molecule. 
     
     
         30 . A pharmaceutical composition comprising the single domain specific binding molecule as claimed in  claim 17 . 
     
     
         31 . The pharmaceutical composition as claimed in  claim 30 , further comprising at least one of a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, a pharmaceutically acceptable adjuvant, and a pharmaceutically acceptable buffer solution. 
     
     
         32 . A method of treating a disease in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition as claimed in  claim 30  to the patient. 
     
     
         33 . A method of imaging a site of disease in a subject, comprising administering the single domain specific binding molecule as claimed in  claim 17  to the subject and imaging a site of disease in the subject. 
     
     
         34 . A method of diagnosing a disease or medical condition in a subject, comprising administering the single domain specific binding molecule as claimed in  claim 17  to the subject and analyzing a sample obtained from the subject.