Antibodies to carcinoembryonic antigen-related cell adhesion molecule (ceacam)

Abstract

The present invention provides antibodies, as well as molecules having at least the antigen-binding portion of an antibody, recognizing a specific epitope of the protein CEACAM1 and optionally binds also other subtypes of the CEACAM protein family. Disclosed antibodies and antibody fragments are characterized by specific CDR sequences. Methods of production and use in therapy and diagnosis, of such antibodies and antibody fragments are also provided.

1 . A monoclonal antibody or an antibody fragment thereof which recognizes human CEACAM1 having a heavy-chain CDR1 comprising a sequence set forth in SEQ ID NO: 1, a heavy-chain CDR2 comprising a sequence set forth in SEQ ID NO: 2 a heavy-chain CDR3 comprising a sequence set forth in SEQ ID NO: 3, a light-chain CDR1 comprising a sequence set forth in SEQ ID NO: 4, a light-chain CDR2 comprising a sequence set forth in SEQ ID NO: 5 and a light-chain CDR3 comprising a sequence set forth in SEQ ID NO: 6, and analogs and derivatives thereof. 
     
     
         2 . The monoclonal antibody or fragment thereof according to  claim 1 :
 (i) comprising heavy chain CDR1 having the sequence set forth in SEQ ID NO: 7, heavy chain CDR2 having the sequence set forth in SEQ ID NO: 8 and heavy chain CDR3 having the sequence set forth in SEQ ID NO: 9;   (ii) comprising heavy chain CDR1 having the sequence set forth in SEQ ID NO: 13, heavy chain CDR2 having the sequence set forth in SEQ ID NO: 14 and heavy chain CDR3 having the sequence set forth in SEQ ID NO: 15;   (iii) comprising light chain CDR1 having the sequence set forth in SEQ ID NO: 10, light chain CDR2 having the sequence set forth in SEQ ID NO: 11 and light chain CDR3 having the sequence set forth in SEQ ID NO: 12;   (iv) comprising light chain CDR1 having the sequence set forth in SEQ ID NO: 16, light chain CDR2 having the sequence set forth in SEQ ID NO: 17, and light chain CDR3 having the sequence set forth in SEQ ID NO: 18;   (v) having CDR sequences set forth in SEQ ID NOs: 13, 14, 15, 16, 17, and 18;   (vi) having CDR sequences set forth in SEQ ID NOs: 7, 8, 9, 10, 11, and 12;   (vii) comprising a heavy chain variable domain sequence having a sequence set forth in SEQ ID NO: 26, or an analog or derivative thereof having at least 97% sequence identity with said heavy chain sequence;   (viii) comprising a light chain variable domain sequence having a sequence set forth in SEQ ID NO: 28, or an analog or derivative thereof having at least 97% sequence identity with said light chain sequence;   (ix) comprising a heavy chain variable domain having a sequence set forth in SEQ ID NO: 26 and a light chain variable domain having a sequence set forth in SEQ ID NO: 28, or an analog or derivative thereof having at least 97% sequence identity with the antibody or fragment sequence; or   (x) Comprising a framework sequence selected from the group consisting of: mouse IgG2a, mouse IgG2b, mouse IgG3, human IgG, human IgG2, human IgG3; and six CDRs having sequences set forth in SEQ ID NOs: 13, 14, 15, 16, 17, and 18; or six CDRs having sequences set forth in SEQ ID NOs: 7, 8, 9, 10, 11, and 12; and analogs and derivatives thereof having at least 97% sequence identity with said CDR sequences, wherein the monoclonal antibody or fragment binds with an affinity of at least about 5×10 −7 M to at least two CEACAM subtypes.   
     
     
         3 . A monoclonal antibody which recognizes CEACAM1, or a fragment thereof comprising at least the antigen-binding portion, which is capable of binding the same epitope on the CEACAM1 molecule to which a monoclonal antibody having the six CDR sequences set forth in SEQ ID NOs: 7, 8, 9, 10, 11 and 12, or the six CDR sequences set forth in SEQ ID NOs: 13, 14, 15, 16, 17 and 18, binds. 
     
     
         4 . The monoclonal antibody or fragment thereof of  claim 3 , wherein the antibody is reactive with an epitope within residues 17-29 and 68-79 of human CEACAM1 having the sequences VLLLVHNLPQQLF (SEQ ID NO:32) and YPNASLLIQNVT (SEQ ID NO:33) respectively. 
     
     
         5 . The monoclonal antibody or fragment thereof according to  claim 4 , wherein:
 (i) the epitope on the CEACAM1 molecule to which the monoclonal antibody binds is an epitope comprising amino acid residues within the sequences VLLLVHNLPQQLF (SEQ ID NO: 32) and YPNASLLIQNVT (SEQ ID NO: 33);   (ii) the epitope on the CEACAM1 molecule to which the monoclonal antibody binds comprises at least four amino acids of the sequence VLLLVHNLPQQLF (SEQ ID NO: 32); or   (iii) the epitope on the CEACAM1 molecule to which the monoclonal antibody binds is na epitope within sequences VLLLVHNLPQQLF (SEQ ID NO: 32) and PNASLLI (SEQ ID NO: 34).   
     
     
         6 . An isolated polynucleotide encoding a monoclonal antibody or antibody fragment according to  claim 1 . 
     
     
         7 . A plasmid comprising at least one isolated polynucleotide sequence according to  claim 6 . 
     
     
         8 . A pharmaceutical composition comprising a therapeutically effective amount of a monoclonal antibody or fragment according to  claim 1 ; and a pharmaceutically acceptable carrier. 
     
     
         9 . A diagnostic composition comprising at least one monoclonal antibody or antibody fragment according to  claim 1 ; and an optional carrier or excipient. 
     
     
         10 . A method of preventing, attenuating or treating a disease or disorder associated with expression, activation or function of CEACAM, comprising administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutical composition according to  claim 8 . 
     
     
         11 . The method of  claim 10 , wherein the disease or disorder is a cancer. 
     
     
         12 . The method of  claim 10  wherein the disease or disorder is a viral infection. 
     
     
         13 . The method according to  claim 10 , wherein the isolated antibody or antibody fragment contained in the pharmaceutical composition is attached to a cytotoxic moiety and the disease is cancer. 
     
     
         14 . The method of  claim 10 , further comprising administering to the subject CEACAM1-expressing lymphocytes. 
     
     
         15 . The method of  claim 14  wherein the lymphocytes comprise T cells, NK cells or Tumor Infiltrating Lymphocyte. 
     
     
         16 . A method of immunomodulation, the method comprising contacting a CEACAM-expressing lymphocyte with the antibody or antibody fragment according to  claim 1 . 
     
     
         17 . A method of inhibiting migration of a CEACAM expressing tumor cell, the method comprising contacting the CEACAM expressing tumor cell with the antibody or antibody fragment according to  claim 1 , thereby inhibiting migration of a CEACAM expressing tumor cell. 
     
     
         18 . A method of inhibiting CEACAM homotypic or heterotypic protein-protein interaction, the method comprising contacting a CEACAM1-expressing lymphocyte with the antibody or antibody fragment according to  claim 1 , thereby inhibiting CEACAM1 homotypic or heterotypic protein-protein interaction. 
     
     
         19 . A method for increasing the duration or progression of response or survival of a subject having cancer, comprising administering to the subject effective amounts of a composition comprising a monoclonal antibody or antibody fragment according to  claim 1 , and an anti-neoplastic composition, wherein said anti-neoplastic composition comprises at least one chemotherapeutic agent, whereby the co-administration of the antibody and the anti-neoplastic composition effectively increases the duration or progression of survival. 
     
     
         20 . A method for diagnosing a cancer in a subject in need thereof, the method comprising contacting a biological sample derived from the subject with the diagnostic composition of  claim 9 , wherein a complex formation beyond a predetermined threshold is indicative of the cancer in the subject. 
     
     
         21 . A method for diagnosing a disease or disorder associated with CEACAM expression, comprising the steps of:
 i. incubating a biological sample with a monoclonal antibody or antibody fragment according to  claim 1 ;   ii. detecting the bound CEACAM using a detectable probe;   iii. comparing the amount of (ii) to a standard curve obtained from reference samples containing known amounts of CEACAM;   iv. calculating the amount of the CEACAM in the biological sample from the standard curve; and   v. comparing the amount of (iv) to a normal CEACAM amount.   
     
     
         22 . A monoclonal antibody or an antibody fragment thereof according to  claim 1  having:
 a heavy-chain CDR1 selected from a sequence set forth in SEQ ID NO: 7 and 
 a sequence set forth in SEQ ID NO: 13; 
 a heavy-chain CDR2 selected from a sequence set forth in SEQ ID NO: 8 and 
 a sequence set forth in SEQ ID NO: 14; 
 a heavy-chain CDR3 selected from a sequence set forth in SEQ ID NO: 9 and 
 a sequence set forth in SEQ ID NO: 15; 
 a light-chain CDR1 selected from a sequence set forth in SEQ ID NO: 10 and 
 a sequence set forth in SEQ ID NO: 16; 
 a light-chain CDR2 selected from a sequence set forth in SEQ ID NO: 11 and 
 a sequence set forth in SEQ ID NO: 17; and 
 a light-chain CDR3 selected from a sequence set forth in SEQ ID NO: 12 and 
 a sequence set forth in SEQ ID NO: 18.