Use of Anti-Alpha5Beta1 Antibodies to Inhibit Cancer Cell Proliferation

Abstract

The present invention provides methods for direct killing of cancer cells using anti-α5β1 antibodies. Generally, the method comprises contacting a cancer cell that expresses α5β1 on its surface with an anti-α5β1 antibody, and thereby inducing the death of the cancer cell. The methods of the invention may be employed at an early stage of cancer development in a patient to prevent tumor establishment. In addition, the methods may be used to treat previously formed tumors especially in cancer that have not proven susceptible to anti-angiogenesis therapy. The methods may be employed as a combination therapy of anti-α5β1 antibodies together with cancer chemotherapeutic agents or other molecular-based cancer therapeutic agents.

1 . A method of inhibiting the proliferation of a cancer cell in a patient wherein the cancer cell expresses α5β1 integrin on its surface, the method comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition comprising the liquid formulation comprising:
 about 1.0 mg/mL to 15 mg/mL anti-α5β1 antibody;   about 22 mM to 27 mM citrate;   about 145 mM to 165 mM sodium chloride;   about 0.04% to 0.06% polysorbate-80;   and a pH of about 5.5 to 7.5.   
     
     
         2 . The method of  claim 1 , wherein the antibody neutralizes at least one biological activity of α5β1 integrin. 
     
     
         3 . The method of  claim 1 , wherein the anti-α5β1 antibody binds to the same epitope of α6β1 integrin as a second antibody comprising a heavy chain variable sequence of SEQ ID NO:6 and a light chain variable sequence of SEQ ID NO:8. 
     
     
         4 . The method of  claim 1 , wherein the second antibody comprises a Fab fragment. 
     
     
         5 . The method of  claim 1 , wherein the anti-α5β1 antibody competitively inhibits binding of a second antibody, comprising a heavy chain variable sequence of SEQ ID NO:6 and a light chain variable sequence of SEQ ID NO: 8, to α5β1 integrin expressed on the cell surface. 
     
     
         6 . The method of  claim 1 , wherein the anti-α5β1 antibody comprises SEQ ID NOs: 2 and 4, or SEQ ID NOs: 6 and 8; or the Fab fragment of SEQ ID NO: 6, and SEQ ID NO:8. 
     
     
         7 . The method of  claim 1 , wherein the antibody competitively inhibits binding of a second antibody to α5β1 integrin on the cancer cell surface wherein the second antibody comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 2 and 4; or SEQ ID NOs: 6 and 8. 
     
     
         8 . The method of  claim 1 , wherein the cancer cell is selected from the group consisting of a bladder cancer cell, a breast cancer cell, a colon cancer cell, a fibrosarcoma cell, a lung cancer cell, a metastatic melanoma cancer cell, an ovarian cancer cell, a pancreatic cancer cell, a prostate cancer cell, a renal cell carcinoma cell, and a spleen cancer cell. 
     
     
         9 . The method of  claim 8 , wherein the cancer is lung, melanoma, ovarian, pancreatic or renal. 
     
     
         10 . The method of  claim 1 , wherein the therapeutically effective dose is about 10 mg/kg. 
     
     
         11 . The method of  claim 1 , further comprising administering a chemotherapeutic agent to the patient either serially or in combination with the antibody. 
     
     
         12 . The method of  claim 1 , wherein the liquid formulation comprises:
 about 10 mg/mL anti-α5β1 antibody;   about 25 mM citrate:   about 150 mM sodium chloride;   about 0.05% polysorbate-80;   and a pH of about 6.5.   
     
     
         13 . The method of  claim 1 , wherein the antibody comprises complementary determining region (CDR) sequences present in a second antibody comprising a heavy chain variable sequence of SEQ ID NO: 6 and a light chain variable sequence of SEQ ID NO: 8, and a human IgG constant region. 
     
     
         14 . The method of  claim 13 , wherein the IgG constant region is IgG4. 
     
     
         15 . The method of  claim 1 , wherein the antibody comprises 95% of the amino acid sequences identical to the CDR sequences present in SEQ ID NOs: 6 and 8.