Anti-Integrin Immunoconjugates, Methods and Uses

Abstract

The invention relates to conjugates of anti-integrin specific antibodies with cytotoxic compounds, the synthesis, selection, and use of such conjugates for use in cancer therapy or other diseases mediated by cell proliferation, cell migration, or inflammation and which pathology involves angiogenesis or neovascularization of new tissue. In addition the invention relates to combination therapy of such diseases wherein the treatment comprises use of said conjugates in combination with one or more other treatment modalities including but not limited to: chemotherapy, surgery or radiation therapy. The preferred conjugates contain maytansinoid compounds linked to the antibody by a disulfide linkage, and preferred chemotherapeutic agents are doxorubicin, a taxane, a camptothecin, a podophyllotoxin, a nucleoside analog, or a pyrimidine analog.

What is claimed is: 
     
         1 . An antibody-drug conjugate of the formula: 
       
         
           
           
               
               
           
         
         wherein the antibody is a monoclonal antibody, or antigen-binding fragment thereof, that competes with CNTO 95 for binding human αV integrin; and wherein the maytansinol is esterified at C-3; R 1  and R 2  are Me; X 1  and X 2  are H; n is 2; p is 2; and m is 3-4, and the pharmaceutically acceptable salts and esters thereof. 
       
     
     
         2 . The antibody conjugate of  claim 1 , wherein the antibody binds an epitope not shared by one or more of the antibodies m7E3 IgG; anti-αVβ3, clone LM609; anti-αVβ5, clone P1F6; anti-β3; or anti-alphaV, clone VNR139. 
     
     
         3 . The antibody conjugate of  claim 1 , wherein the antibody, or antigen-binding fragment thereof, has the epitopic specificity of CNTO 95. 
     
     
         4 . The antibody conjugate of  claim 1 , wherein the antibody, or antigen-binding fragment thereof, comprises the six complementarity determining region (CDR) amino acid sequences of CNTO 95 as shown in SEQ ID NOs: 1-6, or six CDR amino acid sequences with conservative sequence modifications to one or more of the sequences as shown in SEQ ID NOs: 1-6. 
     
     
         5 . The antibody conjugate of  claim 1 , wherein the antibody, or antigen-binding fragment thereof, comprises the six CDRs of CNTO 95 as shown in SEQ ID NOs: 1-6. 
     
     
         6 . The antibody-drug conjugate of  claim 1 , wherein the antibody is a humanized or chimeric antibody. 
     
     
         7 . A method of production of an antibody-drug conjugate according to  claim 1 , comprising the steps: (a) introducing one or more free or protected thiol groups into the antibody by treating the antibody with a bifunctional coupling reagent represented by one of the following structural formulas: 
       
         
           
           
               
               
           
         
       
       to form an activated antibody; (b) reacting the activated antibody with DM4 to form the antibody-drug conjugate; and (c) recovering the antibody-drug conjugate. 
     
     
         8 . A method of treatment of cancer in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a conjugate according to  claim 1 , wherein the cancer expresses αV integrin on its cell surface. 
     
     
         9 . The method of  claim 8 , wherein the cancer is melanoma, lung cancer, or colon cancer. 
     
     
         10 . A method for inhibiting the growth of cancer cells in a mammal in need thereof comprising administering to the mammal a monoclonal antibody conjugate according to  claim 1  in an amount effective to inhibit the growth of said cancer cells in said mammal, wherein the cancer expresses αV integrin on its cell surface. 
     
     
         11 . The method of  claim 10 , wherein the cancer is melanoma, lung cancer, or colon cancer. 
     
     
         12 . The method according to  claim 9 , in which the antibody conjugate is administered intravenously. 
     
     
         13 . The method according to  claim 12 , in which the antibody conjugate is administered in the amount of from about 0.05 mg/kg to about 12.0 mg/kg body weight. 
     
     
         14 . The method according to  claim 10 , in which the mammal is a human. 
     
     
         15 . A pharmaceutical composition comprising a conjugate according to  claim 1  and a pharmaceutically acceptable carrier, diluent, or excipient. 
     
     
         16 . An article of manufacture comprising the conjugate composition of  claim 1  and a container, and further comprising a package insert or label indicating that the composition can be used to treat a disease characterized by cells expressing αV integrin. 
     
     
         17 . An article of manufacture according to  claim 16 , wherein the container comprises a package insert that indicates that the composition can be used to treat cancer. 
     
     
         18 . An antibody conjugate according to  claim 1 , wherein at least one of the heavy chain variable region sequences of the antibody comprises the heavy chain variable region sequence of CNTO 95 as shown in SEQ ID NO: 7. 
     
     
         19 . An antibody conjugate according to  claim 1 , wherein at least one of the light chain variable region sequences of the antibody comprises the light chain variable region sequence of CNTO 95 as shown in SEQ ID NO: 8. 
     
     
         20 . An antibody conjugate according to  claim 1 , wherein at least one of the heavy chain variable region sequences of the antibody comprises the heavy chain variable region sequence of CNTO 95 as shown in SEQ ID NO: 7 and at least one of the light chain variable region sequences of the antibody comprises the light chain variable region sequence of CNTO 95 as shown in SEQ ID NO: 8.