Blood-brain-barrier dual variable domain immunoglobulins and uses thereof

Abstract

The present invention relates to engineered multivalent and multispecific binding proteins, methods of making, and specifically to their uses in the prevention, diagnosis, and/or treatment of disease.

1 . A binding protein comprising a polypeptide chain, wherein said polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein;
 VD1 is a first heavy chain variable domain;   VD2 is a second heavy chain variable domain;   C is a heavy chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 is an Fc region; and   n is 0 or 1;   
       wherein the binding protein is capable of binding a pair of antigens and is able to cross the blood brain barrier (BBB). 
     
     
         2 . The binding protein according to  claim 1 , the binding protein binds a BBB antigen selected from the group consisting of: insulin receptor, transferrin receptor, LRP, and RAGE. 
     
     
         3 . A binding protein comprising a polypeptide chain, wherein said polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein;
 VD1 is a first light heavy chain variable domain;   VD2 is a second light heavy chain variable domain;   C is a light chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 does not comprise an Fc region; and   n is 0 or 1;   
       wherein the binding protein is capable of binding a pair of antigens and is able to cross the blood brain barrier (BBB). 
     
     
         4 . The binding protein according to  claim 3 , wherein the binding protein binds a BBB antigen selected from the group consisting of: insulin receptor, transferrin receptor, LRP, and RAGE. 
     
     
         5 . (canceled) 
     
     
         6 . A binding protein comprising first and second polypeptide chains, wherein said first polypeptide chain comprises a first VD1-(X1)n-VD2-C-(X2)n, wherein
 VD1 is a first heavy chain variable domain;   VD2 is a second heavy chain variable domain;   C is a heavy chain constant domain;   X1 is a linker with the proviso that it is not CH1; and   X2 is an Fc region; and   wherein said second polypeptide chain comprises a second VD1-(X1)n-VD2-C-(X2)n, wherein   VD1 is a first light chain variable domain;   VD2 is a second light chain variable domain;   C is a light chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 does not comprise an Fc region; and   n is 0 or 1, wherein the binding protein is capable of binding a pair of antigens and crosses the blood brain barrier (BBB).   
     
     
         7 . The binding protein according to  claim 6 , wherein the binding protein binds a BBB antigen selected from the group consisting of: insulin receptor, transferrin receptor, LRP, and RAGE. 
     
     
         8 - 9 . (canceled) 
     
     
         10 . The binding protein according to  claim 6 , wherein the Fc region is selected from the group consisting of native sequence Fc region and a variant sequence Fc region. 
     
     
         11 - 19 . (canceled) 
     
     
         20 . The binding protein according to  claim 6 , wherein said first parent antibody or antigen binding portion thereof, and said second parent antibody or antigen binding portion thereof, are selected from the group consisting of: a human antibody; a CDR grafted antibody; a humanized antibody; a Fab fragment; a F(ab′) 2  fragment; a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a dAb fragment; an isolated complementarity determining region (CDR); a single chain antibody; and a diabody. 
     
     
         21 - 23 . (canceled) 
     
     
         24 . A binding protein capable of binding two antigens comprising four polypeptide chains, wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein
 VD1 is a first heavy chain variable domain;   VD2 is a second heavy chain variable domain;   C is a heavy chain constant domain;   X1 is a linker with the proviso that it is not CH1; and   X2 is an Fc region; and   wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein   VD1 is a first light chain variable domain;   VD2 is a second light chain variable domain;   C is a light chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 does not comprise an Fc region; and   n is 0 or 1;   wherein the binding protein crosses the blood brain barrier (BBB).   
     
     
         25 - 26 . (canceled) 
     
     
         27 . The binding protein according to  claim 24 , wherein said binding protein binds a BBB antigen selected from the group consisting of: insulin receptor, transferrin receptor, LRP, and RAGE. 
     
     
         28 . (canceled) 
     
     
         29 . A binding protein conjugate comprising a binding protein according to  claim 24 , wherein said binding protein conjugate further comprising an agent selected from the group consisting of; an immunoadhesion molecule, an imaging agent, a therapeutic agent, and a cytotoxic agent. 
     
     
         30 - 32 . (canceled) 
     
     
         33 . The binding protein according to  claim 24 , wherein said binding protein is a crystallized binding protein. 
     
     
         34 - 36 . (canceled) 
     
     
         37 . An isolated nucleic acid encoding a binding protein amino acid sequence according to  claim 24 . 
     
     
         38 . A vector comprising an isolated nucleic acid according to  claim 37 . 
     
     
         39 . (canceled) 
     
     
         40 . A host cell comprising a vector according to  claim 38 . 
     
     
         41 - 50 . (canceled) 
     
     
         51 . A method of producing a binding protein, comprising culturing a host cell described in  claim 40  in culture medium under conditions sufficient to produce the binding protein 
     
     
         52 - 54 . (canceled) 
     
     
         55 . A protein produced according to the method of  claim 51 . 
     
     
         56 . A pharmaceutical composition comprising the binding protein of  claim 6 , and a pharmaceutically acceptable carrier. 
     
     
         57 - 58 . (canceled) 
     
     
         59 . A method for treating a subject for a disease or a disorder by administering to the subject the binding protein of  claim 6  such that treatment is achieved. 
     
     
         60 . (canceled) 
     
     
         61 . (canceled) 
     
     
         62 . A method for generating a Dual Variable Domain Immunoglobulin capable of binding two antigens comprising the steps of
 a) obtaining a first parent antibody or antigen binding portion thereof, capable of binding a first antigen;   b) obtaining a second parent antibody or antigen binding portion thereof, capable of binding a second antigen;   c) constructing first and third polypeptide chains according to  claim 24  comprising VD1-(X1)n-VD2-C-(X2)n, wherein   VD1 is a first heavy chain variable domain obtained from said first parent antibody or antigen binding portion thereof;   VD2 is a second heavy chain variable domain obtained from said second parent antibody or antigen binding portion thereof;   C is a heavy chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 is an Fc region; and   n is 0 or 1; and   d) constructing second and fourth polypeptide chains according to  claim 24  comprising VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain obtained from said first parent antibody or antigen binding portion thereof;   VD2 is a second light chain variable domain obtained from said second parent antibody or antigen binding thereof;   C is a light chain constant domain;   X1 is a linker with the proviso that it is not CH1;   X2 does not comprise an Fc region; and   n is 0 or 1; and   e) expressing said first, second, third and fourth polypeptide chains;   
       such that a Dual Variable Domain Immunoglobulin capable of binding said first and said second antigen is generated, wherein the binding protein is capable of binding a pair of antigens, wherein the binding protein crosses the blood brain barrier (BBB). 
     
     
         63 . The method of  claim 62 , wherein the binding protein binds a BBB antigen selected from the group consisting of: insulin receptor, transferrin receptor, LRP, and RAGE. 
     
     
         64 . (canceled) 
     
     
         65 . The method of  claim 62 , wherein said first parent antibody or antigen binding portion thereof, and said second parent antibody or antigen binding portion thereof, are selected from the group consisting of: a human antibody; a CDR grafted antibody; a humanized antibody; a Fab fragment, a F(ab′) 2  fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, a dAb fragment, an isolated complementarity determining region (CDR), a single chain antibody, and diabodies. 
     
     
         66 - 76 . (canceled)