Bi-Specific Monovalent Fc Diabodies That Are Capable of Binding CD32B and CD79b and Uses Thereof

Abstract

The present invention is directed to bi-specific monovalent diabodies that comprise an immunoglobulin Fc Domain (“bi-specific monovalent Fc diabodies”) and are composed of three polypeptide chains and which possess at least one binding site specific for an epitope of CD32B and one binding site specific for an epitope of CD79b (i.e., a “CD32B×CD79b bi-specific monovalent Fc diabody”). The bi-specific monovalent Fc diabodies of the present invention are capable of simultaneous binding to CD32B and CD79b. The invention is directed to such compositions, to pharmaceutical compositions that contain such bi-specific monovalent Fc diabodies and to methods for their use in the treatment of inflammatory diseases or conditions, and in particular, systemic lupus erythematosus (SLE) and graft vs. host disease.

What is claimed is: 
     
         1 . A method of treating an inflammatory disease or condition comprising administering an effective amount of a bispecific monovalent Fc diabody to a subject in need thereof,
 wherein said bi-specific monovalent Fc diabody is provided in unit dosage form of at least 5 μg, at least 10 μg, at least 15 μg, at least 25 μg, at least 50 μg, at least 100 μg, or at least 200 μg;   wherein said bi-specific monovalent Fc diabody is capable of specific binding to an epitope of CD32B and to an epitope of CD79b, and possesses an IgG Fc Domain, wherein said bi-specific monovalent Fc diabody comprises a first polypeptide chain, a second polypeptide chain and a third polypeptide chain, wherein said first and second polypeptide chains are covalently bonded to one another and said first and third polypeptide chains are covalently bonded to one another, and wherein:   
       A. the first polypeptide chain comprises, in the N-terminal to C-terminal direction:
 i. a Domain 1, comprising:
 (1) a sub-Domain (1A), which comprises a cysteine-containing peptide (SEQ ID NO:1); and 
 (2) a sub-Domain (1B), which comprises a polypeptide portion of an IgG Fc Domain having CH2 and CH3 domains of an IgG immunoglobulin Fc region; 
 
 ii. a Domain 2, comprising:
 (1) a sub-Domain (2A), which comprises a VL Domain of a monoclonal antibody capable of binding to CD32B (VL CD32B ) (SEQ ID NO:11); and 
 (2) a sub-Domain (2B), which comprises a VH Domain of a monoclonal antibody capable of binding to CD79b (VH CD79b ) (SEQ ID NO:14), 
 wherein said sub-Domains (2A) and (2B) are separated from one another by a peptide linker (SEQ ID NO:4); 
 
 iii. a Domain 3, wherein said Domain 3 is an E-coil Domain (SEQ ID NO:7) or a K-coil Domain (SEQ ID NO:8), wherein said Domain 3 is separated from said Domain 2 by a peptide linker (SEQ ID NO:5); and 
 iv. a C-terminal spacer peptide (SEQ ID NO:6); 
 
       B. the second polypeptide chain comprises, in the N-terminal to C-terminal direction:
 i. a Domain 1, comprising:
 (1) a sub-Domain (1A), which comprises a VL Domain of a monoclonal antibody capable of binding to CD79b (VL CD79b ) (SEQ ID NO:13); and 
 (2) a sub-Domain (1B), which comprises a VH Domain of a monoclonal antibody capable of binding to CD32B (VH CD32B ) (SEQ ID NO:12); 
 wherein said sub-Domains (1A) and (1B) are separated from one another by a peptide linker (SEQ ID NO:4); 
 
 ii. a Domain 2, wherein said Domain 2 is a K-coil Domain (SEQ ID NO:8) or an E-coil Domain (SEQ ID NO:7), 
 wherein said Domain 2 is separated from said Domain 1 by a peptide linker (SEQ ID NO:5); and wherein said Domain 3 of said first polypeptide chain and said Domain 2 of said second polypeptide chain are not both E-coil Domains or both K-coil Domains; and 
 
       C. the third polypeptide chain comprises, in the N-terminal to C-terminal direction, a Domain 1 comprising:
 (1) a sub-Domain (1A), which comprises a cysteine-containing peptide (SEQ ID NO:1); and 
 (2) a sub-Domain (1B), which comprises a polypeptide portion of an IgG Fc Domain having CH2 and CH3 domains of an IgG immunoglobulin Fc region; 
 
       and wherein: 
       (a) said polypeptide portions of the IgG Fc Domains of said first and third polypeptide chains form said IgG Fc Domain; 
       (b) said VL Domain of said first polypeptide chain and said VH Domain of said second polypeptide chain form an Antigen-Binding Domain capable of specific binding to an epitope of CD32B; and 
       (c) said VH Domain of said first polypeptide chain and said VL Domain of said second polypeptide chain form an Antigen-Binding Domain capable of specific binding to an epitope of CD79b. 
     
     
         2 . The method of  claim 1 , wherein said sub-Domain (1B) of said first polypeptide chain comprises a sequence different from that of said sub-Domain (1B) of said third polypeptide chain. 
     
     
         3 . The method of  claim 1 , wherein said sub-Domain (1B) of said first polypeptide chain comprises the amino acid sequence of SEQ ID NO:9, and said sub-Domain (1B) of said third polypeptide chain comprises the amino acid sequence of SEQ ID NO:10. 
     
     
         4 . The method of  claim 1 , wherein said sub-Domain (1B) of said first polypeptide chain comprises the amino acid sequence of SEQ ID NO:10, and said sub-Domain (1B) of said third polypeptide chain comprises the amino acid sequence of SEQ ID NO:9. 
     
     
         5 . The method of  claim 1 , wherein said Domain 1 of said first polypeptide chain and/or said Domain 1 of said third polypeptide chain comprises a variant CH2-CH3 sequence that exhibits altered binding to an Fcγ receptor. 
     
     
         6 . The method of  claim 2 , wherein said Domain 1 of said first polypeptide chain and/or said Domain 1 of said third polypeptide chain comprises a variant CH2-CH3 sequence that exhibits altered binding to an Fcγ receptor. 
     
     
         7 . The method of  claim 1 , wherein said Domain 3 of said first polypeptide chain comprises an E-coil (SEQ ID NO:7), and said Domain 2 of said second polypeptide chain comprises a K-coil (SEQ ID NO:8). 
     
     
         8 . The method of  claim 1 , wherein said Domain 3 of said first polypeptide chain comprises a K-coil (SEQ ID NO:8), and said Domain 2 of said second polypeptide chain comprises an E-coil (SEQ ID NO:7). 
     
     
         9 . The method of  claim 1 , wherein the unit dosage form is administered to the subject at least about 0.01 μg/kg, at least about 0.05 μg/kg, at least about 0.1 μg/kg, at least about 0.2 μg/kg, at least about 0.5 μg/kg, at least about 1 μg/kg, at least about 2 μg/kg, at least about 5 μg/kg, at least about 10 μg/kg, at least about 20 μg/kg, at least about 50 μg/kg, at least about 0.1 mg/kg, at least about 1 mg/kg, at least about 5 mg/kg, at least about 10 mg/kg, at least about 30 mg/kg, at least about 50 mg/kg, at least about 75 mg/kg, at least about 100 mg/kg, at least about 125 mg/kg, at least about 150 mg/kg or more of the subject's body weight. 
     
     
         10 . The method of  claim 1 , wherein the unit dosage form is administered once a week, twice a week, once every two weeks, once a month, once every six weeks, once every two months, twice a year or once per year. 
     
     
         11 . The method of  claim 1 , wherein said inflammatory disease or condition is an autoimmune disease. 
     
     
         12 . The method of  claim 11 , wherein said autoimmune disease is systemic lupus erythematosus. 
     
     
         13 . The method of  claim 1 , wherein said inflammatory disease or condition is graft vs. host disease. 
     
     
         14 . A method of treating an inflammatory disease or condition comprising administering an effective amount of a bispecific monovalent Fc diabody to a subject in need thereof,
 wherein said bi-specific monovalent Fc diabody is provided in unit dosage form of at least 5 μg, at least 10 μg, at least 15 μg, at least 25 μg, at least 50 μg, at least 100 μg, or at least 200 μg;   wherein said bi-specific monovalent Fc diabody is capable of specific binding to an epitope of CD32B and to an epitope of CD79b, and possesses an IgG Fc Domain, wherein said bi-specific monovalent Fc diabody comprises:   
       (1) a first polypeptide chain comprising the amino acid sequence of SEQ ID NO:15; 
       (2) a second polypeptide chain comprising the amino acid sequence of SEQ ID NO:16; and 
       (3) a third polypeptide chain comprising the amino acid sequence of SEQ ID NO:17, wherein amino acid residues 1-10 of said third polypeptide chain are Peptide 1 (SEQ ID NO:1), and amino acid residues 11-227 of said third polypeptide chain are the CH2 and CH3 domains of an IgG antibody Fc region (SEQ ID NO:10); 
       wherein said first and said second polypeptide chains are covalently bonded to one another by a first disulfide bond and said first and third polypeptide chains are covalently bonded to one another by a second disulfide bond. 
     
     
         15 . The method of  claim 14 , wherein the unit dosage form is administered to the subject at least about 0.01 μg/kg, at least about 0.05 μg/kg, at least about 0.1 μg/kg, at least about 0.2 μg/kg, at least about 0.5 μg/kg, at least about 1 μg/kg, at least about 2 μg/kg, at least about 5 μg/kg, at least about 10 μg/kg, at least about 20 μg/kg, at least about 50 μg/kg, at least about 0.1 mg/kg, at least about 1 mg/kg, at least about 5 mg/kg, at least about 10 mg/kg, at least about 30 mg/kg, at least about 50 mg/kg, at least about 75 mg/kg, at least about 100 mg/kg, at least about 125 mg/kg, at least about 150 mg/kg or more of the subject's body weight. 
     
     
         16 . The method of  claim 14 , wherein the unit dosage form is administered once a week, twice a week, once every two weeks, once a month, once every six weeks, once every two months, twice a year or once per year. 
     
     
         17 . The method of  claim 14 , wherein said inflammatory disease or condition is an autoimmune disease. 
     
     
         18 . The method of  claim 17 , wherein said autoimmune disease is systemic lupus erythematosus. 
     
     
         19 . The method of  claim 14 , wherein said inflammatory disease or condition is graft vs. host disease.