J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use

Abstract

In one embodiment, a minibody monomer that binds PSMA is provided. The minibody monomer is encoded by a nucleotide sequence comprising, from N-terminus to C-terminus, an scFv sequence that can bind PSMA, an artificial hinge sequence, and a human IgG CH3 sequence. In another embodiment, a CysDB monomer that binds PSMA is provided. The CysDB monomer may be encoded by a nucleotide sequence comprising, from N-terminus to C-terminus, an scFv sequence that can bind PSMA and a cysteine tail. In other embodiments, methods for diagnosing or treating a cancer associated with PSMA expression in a subject are provided.

1 .- 14 . (canceled) 
     
     
         15 . A method for diagnosing a cancer associated with PSMA expression in a subject, the method comprising:
 administering an anti-PSMA minibody or an anti-PSMA cys-diabody that is conjugated to a diagnostic agent and can bind PSMA to a subject having or suspected of having a cancer associated with PSMA expression;   exposing the subject to an imaging method to visualize the labeled minibody or cys-diabody in vivo; and   determining that the subject has a cancer associated with PSMA expression when the labeled minibody or cys-diabody localizes to a tumor site.   
     
     
         16 . The method of  claim 15 , wherein the minibody comprises SEQ ID NO:10 or SEQ ID NO:11. 
     
     
         17 . The method of  claim 15 , wherein cys-diabody, comprises SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14 or SEQ ID NO:15. 
     
     
         18 . The method of  claim 15 , wherein the anti-PSMA minibody or anti-PSMA cys-diabody targets neovasculature of a solid tumor. 
     
     
         19 . The method of  claim 15 , wherein the cancer associated with PSMA expression in a subject is prostate cancer, lung cancer, colorectal cancer, breast cancer, renal cancer, liver cancer, bladder cancer, pancreatic cancer or melanoma. 
     
     
         20 . A method for treating a cancer associated with PSMA expression in a subject, the method comprising:
 administering a therapeutically effective amount of a pharmaceutical composition to the subject, the composition comprising an anti-PSMA minibody or an anti-PSMA cys-diabody.   
     
     
         21 . The method of  claim 20 , wherein the minibody comprises SEQ ID NO:10 or SEQ ID NO:11. 
     
     
         22 . The method of  claim 20 , wherein the cys-diabody, comprises SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14 or SEQ ID NO:15. 
     
     
         23 . The method of  claim 20 , wherein the anti-PSMA minibody or anti-PSMA cys-diabody is conjugated to a therapeutic agent. 
     
     
         24 . The method of  claim 23 , wherein the therapeutic agent is selected from the group consisting of chemotherapeutic agents, therapeutic antibodies and antibody fragments, toxins, radioisotopes, enzymes, nucleases, hormones, immunomodulators, antisense oligonucleotides, chelators, boron compounds, photoactive agents and dyes. 
     
     
         25 . The method of  claim 20 , wherein the anti-PSMA minibody or anti-PSMA cys-diabody targets neovasculature of a solid tumor. 
     
     
         26 . The method of  claim 19 , wherein the cancer associated with PSMA expression in a subject is prostate cancer, lung cancer, colorectal cancer, breast cancer, renal cancer, liver cancer, bladder cancer, pancreatic cancer or melanoma. 
     
     
         27 . The method of  claim 15 , wherein:
 (a) the minibody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 10; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 10; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 10; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 10; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 10; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 10; 
   (b) the minibody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 11; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 11; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 11; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 11; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 11; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 11; 
   (c) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 12; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 12; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 12; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 12; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 12; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 12; 
   (d) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 13; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 13; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 13; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 13; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 13; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 13; 
   (e) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 14; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 14; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 14; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 14; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 14; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 14; or 
   (f) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 15; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 15; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 15; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 15; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 15; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 15. 
   
     
     
         28 . The method of  claim 20 , wherein:
 (a) the minibody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 10; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 10; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 10; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 10; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 10; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 10; 
   (b) the minibody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 11; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 11; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 11; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 11; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 11; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 11; 
   (c) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 12; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 12; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 12; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 12; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 12; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 12; 
   (d) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 13; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 13; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 13; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 13; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 13; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 13; 
   (e) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 14; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 14; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 14; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 14; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 14; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 14; or 
   (f) the cys-diabody comprises:
 a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 15; 
 a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 15; 
 a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 15; 
 a light chain CDR1 that is a CDR1 in SEQ ID NO: 15; 
 a light chain CDR2 that is a CDR2 in SEQ ID NO: 15; and 
 a light chain CDR3 that is a CDR3 in SEQ ID NO: 15. 
   
     
     
         29 . The method of  claim 15 , wherein the minibody comprises:
 the heavy chain CDR1 comprises amino acid residues 51 to 55 of SEQ ID NO: 10,   the heavy chain CDR2 comprises amino acid residues 70 to 86 of SEQ ID NO: 10, and   the heavy chain CDR3 comprises amino acid residues 119 to 124 of SEQ ID NO: 10.   
     
     
         30 . The method of  claim 15 , wherein the minibody comprises:
 the light chain CDR1 comprises amino acid residues 177 to 187 of SEQ ID NO: 10,   the light chain CDR2 comprises amino acid residues 203 to 209 of SEQ ID NO: 10, and   the light chain CDR3 comprises amino acid residues 242 to 250 of SEQ ID NO: 10.   
     
     
         31 . The method of  claim 15 , wherein the minibody comprises:
 the heavy chain CDR1 comprises amino acid residues 51 to 55 of SEQ ID NO: 10),   the heavy chain CDR2 comprises amino acid residues 70 to 86 of SEQ ID NO: 10,   the heavy chain CDR3 comprises amino acid residues 119 to 124 of SEQ ID NO: 10,   the light chain CDR1 comprises amino acid residues 177 to 187 of SEQ ID NO: 10,   the light chain CDR2 comprises amino acid residues 203 to 209 of SEQ ID NO: 10, and   the light chain CDR3 comprises amino acid residues 242 to 250 of SEQ ID NO: 10.   
     
     
         32 . The method of  claim 15 , wherein the minibody comprises:
 the heavy chain CDR1 comprises amino acid residues 46 to 52 of SEQ ID NO: 10,   the heavy chain CDR2 comprises amino acid residues 72 to 77 of SEQ ID NO: 10, and   the heavy chain CDR3 comprises amino acid residues 119 to 224 of SEQ ID NO: 10.   
     
     
         33 . The method of  claim 15 , wherein the minibody comprises:
 the light chain CDR1 comprises amino acid residues 177 to 187 of SEQ ID NO: 10,   the light chain CDR2 comprises amino acid residues 203 to 209 of SEQ ID NO: 10, and   the light chain CDR3 comprises amino acid residues 242 to 250 of SEQ ID NO: 10.   
     
     
         34 . The method of  claim 15 , wherein the minibody comprises:
 the heavy chain CDR1 comprises amino acid residues 46 to 52 of SEQ ID NO: 10,   the heavy chain CDR2 comprises amino acid residues 72 to 77 of SEQ ID NO: 10,   the heavy chain CDR3 comprises amino acid residues 119 to 224 of SEQ ID NO: 10,   the light chain CDR1 comprises amino acid residues 177 to 187 of SEQ ID NO: 10,   the light chain CDR2 comprises amino acid residues 203 to 209 of SEQ ID NO: 10, and   the light chain CDR3 comprises amino acid residues 242 to 250 of SEQ ID NO: 10.