Methods of treatment using ctla4 mutant molecules

Abstract

The present invention provides soluble CTLA4 mutant molecules which bind with greater avidity to the CD80 and/or CD86 antigen than wild type CTLA4 or non-mutated CTLA4Ig. The soluble CTLA4 molecules have a first amino acid sequence comprising the extracellular domain of CTLA4, where certain amino acid residues within the S25-R33 region and M97-G107 region are mutated. The mutant molecules of the invention may also include a second amino acid sequence which increases the solubility of the mutant molecule.

1 . A method for treating graft versus host disease in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule binds CD80 and/or CD86 and comprises an extracellular domain of CTLA4 as shown in SEQ ID NO:8 beginning with alanine at position 26 or methionine at position 27 and ending with aspartic acid at position 150, or a portion thereof, wherein in the extracellular domain or portion thereof, an alanine at position 55 is substituted with a tyrosine, and a leucine at position 130 is substituted with a glutamic acid. 
     
     
         2 . A method for treating graft versus host disease in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
 (a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4.   
     
     
         3 . A method for treating graft versus host disease in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
 (a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4 or a portion thereof that binds CD80 and/or CD86, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4 or a portion thereof that binds CD80 and/or CD86.   
     
     
         4 . The method of  claim 2  further comprising an amino acid sequence which alters the solubility or affinity of the CTLA4 mutant molecule. 
     
     
         5 . The method of  claim 4 , wherein the amino acid sequence which alters the solubility or affinity comprises an immunoglobulin. 
     
     
         6 . The method of  claim 5 , wherein the immunoglobulin is an immunoglobulin constant region or portion thereof. 
     
     
         7 . The method of  claim 6 , wherein the immunoglobulin constant region or portion thereof is mutated to reduce effector function. 
     
     
         8 . The method of  claim 6 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule. 
     
     
         9 . The method of  claim 7 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule. 
     
     
         10 . A method for treating graft versus host disease in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
 (a) an amino acid sequence beginning with methionine at position 27 and ending with lysine at position 383 of SEQ ID NO:4, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with lysine at position 383 of SEQ ID NO:4.   
     
     
         11 . A method for treating graft versus host disease in a subject comprising administering to the subject a CTLA4 mutant molecule encoded by the nucleic acid molecule designated ATCC No. PTA-2104. 
     
     
         12 . The method of  claims 2 , and  10 - 11 , wherein the CTLA4 mutant molecule is administered concomitantly or in sequence with one or more agents selected from the group consisting of corticosteroids, azathioprine, tacrolimus, basiliximab, soluble gp39 (also known as CD40 ligand (CD40L), CD154, T-BAM, TRAP), soluble CD29, soluble CD40, soluble CD80, soluble CD86, soluble CD28, soluble CD56, soluble Thy-1, soluble CD3, soluble TCR, soluble VLA-4, soluble VCAM-1, soluble LECAM-1, soluble ELAM-1, soluble CD44, antibodies reactive with gp39, antibodies reactive with CD40, antibodies reactive with B7, antibodies reactive with CD28, antibodies reactive with LFA-1, antibodies reactive with LFA-2, antibodies reactive with IL-2, antibodies reactive with IL-12, antibodies reactive with IFN-gamma, antibodies reactive with CD2, antibodies reactive with CD48, antibodies reactive with any ICAM (e.g., ICAM-2), antibodies reactive with CTLA4, antibodies reactive with Thy-1, antibodies reactive with CD56, antibodies reactive with CD3, antibodies reactive with CD29, antibodies reactive with TCR, antibodies reactive with VLA-4, antibodies reactive with VCAM-1, antibodies reactive with LECAM-1, antibodies reactive with ELAM-1, antibodies reactive with CD44, a calcineurin inhibitor, e.g. cyclosporin A or FK506; an immunosuppressive macrolide, e.g. rapamycine or a derivative thereof; e.g. 40-O-(2-hydroxy)ethyl-rapamycin, a lymphocyte horning agent, e.g. FTY720 or an analog thereof; cyclophosphamide; methotrexate; leflunomide or an analog thereof; mizoribine; mycophenolic acid; mycophenolate mofetil; 15-deoxyspergualine or an analog thereof; immunosuppressive monoclonal antibodies, e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3, CD4, CD 11a/CD18, CD7, CD25, CD 27, B7, CD40, CD45, CD58, CD 137, ICOS, CD150 (SLAM), OX40, 4-1BB or their ligands; or other immunomodulatory compounds, e.g. CTLA4/CD28-Ig, or other adhesion molecule inhibitors, e.g. mAbs or low molecular weight inhibitors including LFA-1 antagonists, Selectin antagonists and VLA-4 antagonists. 
     
     
         13 . The method of treating an autoimmune disease in a subject comprising administering to the subject a CTLA4 molecule, wherein the CTLA4 molecule comprises:
 (a) an amino acid sequence beginning with, methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:8, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:8, and   
       wherein said autoimmune disease is selected from the group consisting of psoriasis, Hashimoto's thyroiditis, primary myxedema, Graves' disease, pernicious anemia, autoimmune atrophic gastritis, Addison's disease, diabetes, insulin dependent diabetes mellitis, type I diabetes mellitis, good pasture's syndrome, myasthenia gravis, pemphigus, Crohn's disease, sympathetic ophthalmia, autoimmune uveitis, multiple sclerosis, autoimmune hemolytic anemia, idiopathic thrombocytopenia, primary biliary cirrhosis, chronic action hepatitis, ulceratis colitis, Sjogren's syndrome, polymyositis, scleroderma, and mixed connective tissue disease. 
     
     
         14 . The method of treating an immunoproliferative disease in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
 (a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4, and   
       wherein said immunoproliferative disease is selected from the group consisting of T cell lymphoma, T cell acute lymphoblastic leukemia, testicular angiocentric T cell lymphoma and benign lymphocytic angiitis. 
     
     
         15 . The method of  claim 13  further comprising an amino acid sequence which alters the solubility or affinity of the CTLA4 mutant molecule. 
     
     
         16 . The method of  claim 15 , wherein the amino acid sequence which alters the solubility or affinity comprises an immunoglobulin. 
     
     
         17 . The method of  claim 16 , wherein the immunoglobulin is an immunoglobulin constant region or portion thereof. 
     
     
         18 . The method of  claim 17 , wherein the immunoglobulin constant region or portion thereof is mutated to reduce effector function. 
     
     
         19 . The method of  claim 17 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule. 
     
     
         20 . The method of  claim 18 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule. 
     
     
         21 . The method of  claim 13 , wherein the CTLA4 molecule comprises:
 (a) an amino acid sequence beginning with methionine at position 27 and ending with lysine at position 383 of SEQ ID NO: 8, or   (b) an amino acid sequence beginning with alanine at position 26 and ending with lysine at position 383 of SEQ ID NO: 8.