Novel immunotherapy against several tumors including gastrointestinal and gastric cancer

Abstract

A pharmaceutical composition contains an antibody or a fragment thereof specific for COL6A3 for the treatment of a cancer. A method of treating a cancer includes administering to a subject in need thereof the pharmaceutical composition. A kit includes a container that contains the pharmaceutical composition. A method of producing an antibody or a fragment thereof against a peptide or a MHC/peptide complex. A method for detecting a diseased tissue includes administering to a subject in need thereof an antibody or a fragment thereof conjugated to a radioisotope and detecting a signal from the radioisotope in the subject. A method for treating a diseased tissue includes administering to a subject in need thereof an antibody or a fragment thereof conjugated to a toxin.

1 . A pharmaceutical composition comprising an antibody or a fragment thereof specific for COL6A3, wherein the antibody or the fragment thereof specifically binds to an epitope of COL6A3 selected from the group consisting of SYLQAANAL (SEQ ID NO: 23), FLLDGSANV (SEQ ID NO: 68), NLLDLDYEL (SEQ ID NO: 69), and FLIDSSEGV (SEQ ID NO: 70). 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the epitope is FLLDGSANV (SEQ ID NO: 68). 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the fragment thereof is an Fab fragment. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the antibody or the fragment thereof is a polyclonal antibody, a monoclonal antibody, or a humanized chimeric antibody. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the antibody or the fragment thereof is conjugated to a radioisotope and/or a toxin. 
     
     
         6 . The pharmaceutical composition of  claim 1 , further comprises a pharmaceutically acceptable carrier. 
     
     
         7 . A method of treating gastric cancer comprising administering to a subject in need thereof the pharmaceutical composition of  claim 1 . 
     
     
         8 . The method of  claim 7 , wherein the administering is by intravenous, intraperitoneal, subcutaneous, or intramuscular injection. 
     
     
         9 . A kit comprising
 (a) a container comprising the pharmaceutical composition of  claim 1  in solution or in lyophilized form;   (b) optionally a second container containing a diluent or reconstituting solution for the lyophilized formulation; and   (c) optionally, instructions for (i) use of the solution or (ii) reconstitution and/or use of the lyophilized formulation.   
     
     
         10 . The kit of  claim 9 , further comprises one or more of (iii) a buffer, (iv) a diluent, (v) a filter, (vi) a needle, or (v) a syringe. 
     
     
         11 . The kit of  claim 9 , wherein the pharmaceutical composition is in lyophilized form. 
     
     
         12 . A method of producing an antibody or a fragment thereof of  claim 1 , comprising
 (a) immunizing an animal with the peptide,   (b) obtaining antibody-producing cells from the immunized animal,   (c) fusing tumor cells with the antibody-producing cells to produce hybridomas,   (d) screening the hybridomas to identify a clone that produces the antibody or the fragment thereof specifically binding the peptide,   (e) expanding the clone, and   (f) isolating the antibody or the fragment thereof from the expanded clone,   wherein the peptide is selected from the group consisting of SYLQAANAL (SEQ ID NO: 23), FLLDGSANV (SEQ ID NO: 68), NLLDLDYEL (SEQ ID NO: 69), and FLIDSSEGV (SEQ ID NO: 70).   
     
     
         13 . The method of  claim 12  further comprising
 (g) isolating DNA encoding the antibody or the fragment thereof, 
 (h) expressing the isolated DNA in a host cell, and 
 (i) isolating the antibody or the fragment thereof produced by the host cell. 
 
     
     
         14 . The method of  claim 13 , further comprising
 (j) substituting at least one complementarity-determining region (CDR) sequence of the isolated DNA for the corresponding sequence of a human antibody.   
     
     
         15 . The method of  claim 14 , further comprising
 (k) conjugating the antibody or the fragment thereof to a radioisotope and/or a toxin.   
     
     
         16 . The method of  claim 13 , wherein the epitope is FLLDGSANV (SEQ ID NO: 68). 
     
     
         17 . The method of  claim 13 , wherein the fragment thereof is an Fab fragment. 
     
     
         18 . A method of producing an antibody or a fragment thereof of  claim 1  against a MHC/peptide complex, comprising
 (a) immunizing an animal with the MHC/peptide complex, 
 (b) obtaining antibody-producing cells from the immunized animal, 
 (c) fusing tumor cells with the antibody-producing cells to produce hybridomas, 
 (d) screening the hybridomas to identify a clone that produces the antibody or the fragment thereof specifically binding the MHC/peptide complex, 
 (e) expanding the clone, and 
 (f) isolating the antibody or the fragment thereof from the expanded clone, 
 wherein the peptide is selected from the group consisting of SYLQAANAL (SEQ ID NO: 23), FLLDGSANV (SEQ ID NO: 68), NLLDLDYEL (SEQ ID NO: 69), and FLIDSSEGV (SEQ ID NO: 70). 
 
     
     
         19 . The method of  claim 18 , further comprising
 (g) isolating DNA encoding the antibody or the fragment thereof,   (h) expressing the isolated DNA in a host cell, and   (i) isolating the antibody or the fragment thereof produced by the host cell.   
     
     
         20 . The method of  claim 19 , further comprising
 (j) substituting at least one complementarity-determining region (CDR) sequence of the isolated DNA for the corresponding sequence of a human antibody.   
     
     
         21 . The method of  claim 20 , further comprising
 (k) conjugating the antibody or the fragment thereof to a radioisotope and/or a toxin.   
     
     
         22 . The method of  claim 18 , wherein the peptide is FLLDGSANV (SEQ ID NO: 68). 
     
     
         23 . The method  claim 18 , wherein the fragment thereof is an Fab fragment. 
     
     
         24 . The method of  claim 18 , wherein the MHC is a MHC class-I or class-II molecule. 
     
     
         25 . An antibody or a fragment thereof produced by the method of  claim 12 . 
     
     
         26 . A method for detecting a diseased tissue, comprising
 (a) administering to a subject in need thereof an antibody or a fragment thereof conjugated to a radioisotope produced by the method of  claim 21 , and   (b) detecting a signal from the radioisotope in the subject.   
     
     
         27 . The method of  claim 26 , wherein the detecting is by positron emission tomography (PET). 
     
     
         28 . The method of  claim 26 , wherein the detecting is to determine metastasis or size and location of the diseased tissue. 
     
     
         29 . A method for treating a diseased tissue, comprising administering to a subject in need thereof an antibody or a fragment thereof conjugated to a toxin produced by the method of  claim 21 . 
     
     
         30 . The method of  claim 28 , wherein the diseased tissue is a cancer. 
     
     
         31 . The method of  claim 30 , wherein the cancer is gastric cancer. 
     
     
         32 . The method of  claim 26 , wherein the administering is by intravenous, intraperitoneal, subcutaneous, or intramuscular injection