Multispecific heavy chain antibodies with modified heavy chain constant regions

Abstract

Multispecific, human heavy chain antibodies (e.g., UniAbs™) that have modified heavy chain constant regions that impart advantageous properties are provided. The invention further concerns methods of making such antibodies, compositions, including pharmaceutical compositions, comprising such antibodies, and their use to treat disorders that are characterized by expression of one or more of the binding targets described herein.

1 . An isolated multispecific antibody comprising:
 a first heavy chain polypeptide subunit comprising a mutated human IgG4 constant region comprising mutations S228P, F234A, L235A, and T366W; and   a second heavy chain polypeptide subunit comprising a mutated human IgG4 constant region comprising mutations S228P, F234A, L235A, T366S, L368A, and Y407V.   
     
     
         2 . The isolated multispecific antibody of  claim 1 , wherein the mutated human IgG4 constant region of the first heavy chain polypeptide subunit or the mutated human IgG4 constant region of second heavy chain polypeptide subunit lacks a CH1 domain. 
     
     
         3 . The isolated multispecific antibody of  claim 1  or  2 , wherein the mutated human IgG4 constant region of the first heavy chain polypeptide subunit comprises a sequence of SEQ ID NO: 73 or 55, and the mutated human IgG4 constant region of the second heavy chain polypeptide subunit comprises a sequence of SEQ ID NO: 72 or 54. 
     
     
         4 . The isolated multispecific antibody of any one of  claims 1 - 3 , further comprising a first binding moiety that has binding specificity for CD3, comprising:
 a heavy chain variable domain comprising a CDR1 sequence comprising a sequence of SEQ ID NO: 36, a CDR2 sequence comprising a sequence of SEQ ID NO: 37, and a CDR3 sequence comprising a sequence of SEQ ID NO: 38; and   a light chain variable domain comprising a CDR1 sequence comprising a sequence of SEQ ID NO: 39, a CDR2 sequence comprising a sequence of SEQ ID NO: 40, and a CDR3 sequence comprising a sequence of SEQ ID NO: 41.   
     
     
         5 . The isolated multispecific antibody of  claim 4 , wherein:
 the CDR1, CDR2 and CDR3 sequences in the heavy chain variable domain of the first binding moiety are present in a human VH framework; and   the CDR1, CDR2 and CDR3 sequences in the light chain variable domain of the first binding moiety are present in a human Vkappa framework.   
     
     
         6 . The isolated multispecific antibody of  claim 5 , wherein:
 the heavy chain variable domain of the first binding moiety comprises a sequence having at least 95% identity to SEQ ID NO: 42; and   the light chain variable domain of the first binding moiety comprises a sequence having at least 95% identity to SEQ ID NO: 43.   
     
     
         7 . The isolated multispecific antibody of  claim 6 , wherein:
 the heavy chain variable domain of the first binding moiety comprises the sequence of SEQ ID NO: 42; and   the light chain variable domain of the first binding moiety comprises the sequence of SEQ ID NO: 43.   
     
     
         8 . The isolated multispecific antibody of any one of  claims 1 - 7 , further comprising a second binding moiety having binding specificity to a protein other than CD3. 
     
     
         9 . The isolated multispecific antibody of  claim 8 , wherein the second binding moiety comprises a single heavy chain variable region, in a monovalent or bivalent configuration. 
     
     
         10 . The isolated multispecific antibody of  claim 9 , wherein the first binding moiety comprises a light chain polypeptide subunit and a heavy chain polypeptide subunit, and wherein the second binding moiety comprises a heavy chain polypeptide subunit. 
     
     
         11 . The isolated multispecific antibody of  claim 10 , wherein the light chain polypeptide subunit of the first binding moiety comprises a light chain constant domain. 
     
     
         12 . The isolated multispecific antibody of any one of  claims 8 - 11 , wherein the protein other than CD3 is a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA). 
     
     
         13 . The isolated multispecific antibody of  claim 12 , wherein the TAA is B-cell maturation antigen (BCMA). 
     
     
         14 . The isolated multispecific antibody of  claim 12 , wherein the TAA is CD19. 
     
     
         15 . The isolated multispecific antibody of  claim 12 , wherein the TAA is prostate specific membrane antigen (PSMA). 
     
     
         16 . A pharmaceutical composition comprising a multispecific antibody according to any one of  claims 1 - 15 . 
     
     
         17 . A polynucleotide encoding a multispecific antibody according to any one of  claims 1 - 15 . 
     
     
         18 . A vector comprising a polynucleotide according to  claim 17 . 
     
     
         19 . A cell comprising a vector according to  claim 18 . 
     
     
         20 . A method of producing a multispecific antibody according to any one of  claims 1 - 15 , comprising growing a cell according to  claim 19  under conditions permissive for expression of the multispecific antibody, and isolating the multispecific antibody from the cell. 
     
     
         21 . A method of treatment, comprising administering to an individual in need an effective dose of the multispecific antibody of any one of  claims 1 - 15 , or the pharmaceutical composition of  claim 16 . 
     
     
         22 . Use of a multispecific antibody of any one of  claims 1 - 15  in the preparation of a medicament for the treatment of a disease or disorder in an individual in need. 
     
     
         23 . The multispecific antibody of any one of  claims 1 - 15 , or the pharmaceutical composition of  claim 16 , for use in therapy in an individual in need. 
     
     
         24 . A method for treating a disease or condition characterized by expression of BCMA, comprising administering to an individual in need an effective dose of the multispecific antibody according to  claim 13 , or a pharmaceutical composition comprising the multispecific antibody of  claim 13 . 
     
     
         25 . The method of  claim 24 , wherein the disease is an autoimmune disease. 
     
     
         26 . The method of  claim 24 , wherein the disease is a cancer. 
     
     
         27 . The method of  claim 26 , wherein the cancer is a myeloma. 
     
     
         28 . The method of  claim 27 , wherein the myeloma is multiple myeloma. 
     
     
         29 . A method for treating a disease or condition characterized by expression of PSMA, comprising administering to an individual in need an effective dose of the multispecific antibody according to  claim 15 , or a pharmaceutical composition comprising the multispecific antibody of  claim 15 . 
     
     
         30 . The method of  claim 29 , wherein the disease is a cancer. 
     
     
         31 . The method of  claim 29 , wherein the cancer is prostate cancer. 
     
     
         32 . A method for treating a disease or condition characterized by expression of CD19, comprising administering to an individual in need an effective dose of the multispecific antibody according to  claim 14 , or a pharmaceutical composition comprising the multispecific antibody of  claim 14 . 
     
     
         33 . The method of  claim 32 , wherein the disorder is diffuse large B-cell lymphoma (DLBCL). 
     
     
         34 . The method of  claim 32 , wherein the disorder is acute lymphoblastic leukemia (ALL). 
     
     
         35 . The method of  claim 32 , wherein the disorder is non-Hodgkin's lymphoma (NHL). 
     
     
         36 . The method of  claim 32 , wherein the disorder is systemic lupus erythematosus (SLE). 
     
     
         37 . The method of  claim 32 , wherein the disorder is rheumatoid arthritis (RA). 
     
     
         38 . The method of  claim 32 , wherein the disorder is multiple sclerosis (MS). 
     
     
         39 . A kit for treating a disease or disorder in an individual in need, comprising a multispecific antibody of any one of  claims 1 - 15 , or the pharmaceutical composition of  claim 16 , and instructions for use. 
     
     
         40 . The kit of  claim 39 , further comprising at least one additional reagent. 
     
     
         41 . The kit of  claim 40 , wherein the at least one additional reagent comprises a chemotherapeutic drug. 
     
     
         42 . A bispecific three-chain antibody like molecule comprising:
 a first polypeptide subunit comprising:   a light chain variable domain (VL) comprising a sequence of SEQ ID NO: 43; and   a light chain constant domain (CL);   a second polypeptide subunit comprising:   a heavy chain variable domain (VH) comprising a sequence of SEQ ID NO: 42; and   a heavy chain constant domain (CH) comprising a sequence of SEQ ID NO: 72 or 73;   wherein the light chain variable domain and the heavy chain variable domain together form a first binding moiety that binds to CD3; and   a third polypeptide subunit comprising:   a heavy chain-only variable region, in a monovalent or bivalent configuration, that binds to a protein other than CD3; and   a heavy chain constant domain (CH) comprising a sequence of SEQ ID NO: 54 or 55.   
     
     
         43 . The bispecific three-chain antibody like molecule of  claim 42 , wherein the third polypeptide subunit comprises a heavy chain-only variable region in a bivalent configuration that binds to BCMA. 
     
     
         44 . The bispecific three-chain antibody like molecule of  claim 43 , comprising:
 a first polypeptide subunit comprising a sequence of SEQ ID NO: 49;   a second polypeptide subunit comprising a sequence of SEQ ID NO: 56; and   a third polypeptide subunit comprising a sequence of SEQ ID NO: 58.   
     
     
         45 . A pharmaceutical composition comprising the bispecific three-chain antibody like molecule of any one of  claims 42 - 44 . 
     
     
         46 . A polynucleotide encoding a bispecific three-chain antibody like molecule according to any one of  claims 42 - 44 . 
     
     
         47 . A vector comprising a polynucleotide according to  claim 46 . 
     
     
         48 . A cell comprising a vector according to  claim 47 . 
     
     
         49 . A method of producing a bispecific three-chain antibody like molecule according to any one of  claims 42 - 44 , comprising growing a cell according to  claim 48  under conditions permissive for expression of the bispecific three-chain antibody like molecule, and isolating the bispecific three-chain antibody like molecule from the cell. 
     
     
         50 . A method of treatment, comprising administering to an individual in need an effective dose of the bispecific three-chain antibody like molecule of any one of  claims 42 - 44 , or the pharmaceutical composition of  claim 45 . 
     
     
         51 . Use of a bispecific three-chain antibody like molecule of any one of  claims 42 - 44  in the preparation of a medicament for the treatment of a disease or disorder in an individual in need. 
     
     
         52 . The bispecific three-chain antibody like molecule of any one of  claims 42 - 44 , or the pharmaceutical composition of  claim 45 , for use in therapy in an individual in need. 
     
     
         53 . A method for treating a disease or condition characterized by expression of BCMA, comprising administering to an individual in need an effective dose of the bispecific three-chain antibody like molecule according to any one of  claims 42 - 44 , or the pharmaceutical composition of  claim 45 . 
     
     
         54 . The method of  claim 53 , wherein the disease is an autoimmune disease. 
     
     
         55 . The method of  claim 53 , wherein the disease is a cancer. 
     
     
         56 . The method of  claim 55 , wherein the cancer is a myeloma. 
     
     
         57 . The method of  claim 56 , wherein the myeloma is multiple myeloma. 
     
     
         58 . A kit for treating a disease or disorder in an individual in need, comprising a bispecific three-chain antibody like molecule of any one of  claims 42 - 44 , or the pharmaceutical composition of  claim 45 , and instructions for use. 
     
     
         59 . The kit of  claim 58 , further comprising at least one additional reagent. 
     
     
         60 . The kit of  claim 59 , wherein the at least one additional reagent comprises a chemotherapeutic drug.