Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9

Abstract

The invention relates to new bispecific antigen binding molecules, comprising at least one antigen binding domain capable of specific binding to 4-1BB, at least one moiety capable of specific binding to a target cell antigen, and a Fc domain composed of a first and a second subunit capable of stable association, and to methods of producing these molecules and to methods of using the same.

1 . A bispecific antigen binding molecule, comprising at least one antigen binding domain capable of specific binding to 4-1BB, at least one antigen binding domain capable of specific binding to a target cell antigen and a Fc domain composed of a first and a second subunit capable of stable association,
 wherein the antigen binding domain capable of specific binding to 4-1BB comprises a heavy chain variable region (V H 4-1BB) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:1, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:2, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:3, and a light chain variable region (V L 4-1BB) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:4, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:5, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:6.   
     
     
         2 . The bispecific antigen binding molecule of  claim 1 , wherein the antigen binding domain capable of specific binding to 4-1BB comprises a heavy chain variable region (V H 4-1BB) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:7 and a light chain variable region (V L 4-1BB) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:8. 
     
     
         3 . The bispecific antigen binding molecule of  claim 1 , wherein antigen binding domain capable of specific binding to 4-1BB is a Fab fragment. 
     
     
         4 . The bispecific antigen binding molecule of  claim 1 , wherein the target cell antigen is selected from the group consisting of Fibroblast Activation Protein (FAP), Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Carcinoembryonic Antigen (CEA), CD19, CD20 and CD33. 
     
     
         5 . The bispecific antigen binding molecule of  claim 1 , wherein the antigen binding domain capable of specific binding to a target cell antigen is an antigen binding domain capable of specific binding to Fibroblast Activation Protein (FAP). 
     
     
         6 . The bispecific antigen binding molecule of  claim 5 , wherein the antigen binding domain capable of specific binding to Fibroblast Activation Protein (FAP) comprises
 (a) a heavy chain variable region (V H FAP) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:9, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:10, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:11, and a light chain variable region (V L FAP) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:12, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:13, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:14, or   (b) a heavy chain variable region (V H FAP) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:15, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:16, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, and a light chain variable region (V L FAP) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:18, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:19, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:20.   
     
     
         7 . The bispecific antigen binding molecule of  claim 5 , wherein the antigen binding domain capable of specific binding to Fibroblast Activation Protein (FAP) comprises
 (a) a heavy chain variable region (V H FAP) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:21, and a light chain variable region (V L FAP) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:22, or   (b) a heavy chain variable region (V H FAP) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:23, and a light chain variable region (V L FAP) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:24.   
     
     
         8 . The bispecific antigen binding molecule of  claim 1 , wherein the antigen binding domain capable of specific binding to a target cell antigen is an antigen binding domain capable of specific binding to Carcinoembryonic Antigen (CEA). 
     
     
         9 . The bispecific antigen binding molecule of  claim 8 , wherein the antigen binding domain capable of specific binding to Carcinoembryonic Antigen (CEA) comprises
 (a) a heavy chain variable region (V H CEA) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:25, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:26, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:27, and a light chain variable region (V L CEA) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:28, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:29, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:30, or   (b) a heavy chain variable region (V H CEA) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:33, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:34, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:35, and a light chain variable region (V L CEA) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:36, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:37, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:38, or   (c) a heavy chain variable region (V H CEA) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:179, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:180, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:181, and a light chain variable region (V L CEA) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:182, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:183, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:184, or   (d) a heavy chain variable region (V H CEA) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:187, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:188, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:189, and a light chain variable region (V L CEA) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:190, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:191, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:192.   
     
     
         10 . The bispecific antigen binding molecule of  claim 8 , wherein the antigen binding domain capable of specific binding to Carcinoembryonic Antigen (CEA) comprises (a) a heavy chain variable region (V H CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:31, and a light chain variable region (V L CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:32, or (b) a heavy chain variable region (V H CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:39, and a light chain variable region (V L CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:40, or (c) a heavy chain variable region (V H CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:185, and a light chain variable region (V L CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:186, or (d) a heavy chain variable region (V H CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:193, and a light chain variable region (V L CEA) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:194. 
     
     
         11 . The bispecific antigen binding molecule of  claim 1 , wherein the antigen binding domain capable of specific binding to a target cell antigen is an antigen binding domain capable of specific binding to CD19. 
     
     
         12 . The bispecific antigen binding molecule of  claim 11 , wherein the antigen binding domain capable of specific binding to CD19 comprises
 (a) a heavy chain variable region (V H CD19) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:155, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:156, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:157, and a light chain variable region (V L CD19) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:158, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:159, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:160, or   
       (b) a heavy chain variable region (V H CD19) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:163, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:164, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:165, and a light chain variable region (V L CD19) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:166, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:167, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:168. 
     
     
         13 . The bispecific antigen binding molecule of  claim 11 , wherein the antigen binding domain capable of specific binding to CD19 comprises (a) a heavy chain variable region (V H CD19) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:161, and a light chain variable region (V L CD19) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:162, or (b) a heavy chain variable region (V H CD19) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:169, and a light chain variable region (V L CD19) comprising an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:170. 
     
     
         14 . The bispecific antigen binding molecule of  claim 1 , wherein the bispecific antigen binding molecule comprises an IgG Fc domain. 
     
     
         15 . The bispecific antigen binding molecule of  claim 1 , wherein the Fc domain comprises one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function. 
     
     
         16 . The bispecific antigen binding molecule of  claim 1 , comprising
 (a) two antigen binding domains capable of specific binding to 4-1BB,   (b) one antigen binding domain capable of specific binding to a target cell antigen, and   (c) an IgG Fc domain comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function.   
     
     
         17 . The bispecific antigen binding molecule of  claim 1 , comprising
 (a) two light chains and two heavy chains of an antibody comprising two Fab fragments capable of specific binding to 4-1BB and the Fc domain, and   (b) a VH and VL domain capable of specific binding to a target cell antigen, wherein the VH domain is connected via a peptide linker to the C-terminus of one of the two heavy chains and wherein the VL domain is connected via a peptide linker to the C-terminus of the second heavy chain.   
     
     
         18 . The bispecific antigen binding molecule of  claim 1 , comprising
 (a) two light chains and two heavy chains of an antibody comprising two Fab fragments capable of specific binding to 4-1BB and the Fc domain, and   (b) a crossFab fragment capable of specific binding to the target cell antigen which is connected via a peptide linker to the C-terminus of one of the two heavy chains.   
     
     
         19 . The bispecific antigen binding molecule of  claim 1 , comprising
 (a) four antigen binding domains capable of specific binding to 4-1BB,   (b) one antigen binding domain capable of specific binding to a target cell antigen, and   (c) an IgG Fc domain comprising one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function.   
     
     
         20 . A polynucleotide encoding the bispecific antigen binding molecule of  claim 1 . 
     
     
         21 . A host cell comprising the polynucleotide of  claim 20 . 
     
     
         22 . A method of producing the bispecific antigen binding molecule comprising culturing the host cell of  claim 21  under conditions suitable for the expression of the bispecific antigen binding molecule. 
     
     
         23 . A pharmaceutical composition comprising a bispecific antigen binding molecule of  claim 1  and at least one pharmaceutically acceptable excipient. 
     
     
         24 .- 29 . (canceled) 
     
     
         30 . A method of inhibiting the growth of tumor cells in an individual comprising administering to the individual an effective amount of the bispecific antigen binding molecule of any one of  claim 1 , to inhibit the growth of the tumor cells. 
     
     
         31 . A method of treating cancer or an infectious disease comprising administering to the individual a therapeutically effective amount of the bispecific antigen binding molecule of  claim 1 . 
     
     
         32 . A bispecific antigen binding molecule, comprising at least one antigen binding domain capable of specific binding to mouse 4-1BB, at least one antigen binding domain capable of specific binding to a target cell antigen and a Fc domain composed of a first and a second subunit capable of stable association,
 wherein the antigen binding domain capable of specific binding to 4-1BB comprises a heavy chain variable region (V H 4-1BB) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:171, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:172, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:173, and a light chain variable region (V L 4-1BB) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:174, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:175, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:176.   
     
     
         33 . The bispecific antigen binding molecule of  claim 32 , wherein the antigen binding domain capable of specific binding to a target cell antigen binds to Fibroblast Activation Protein (FAP) and comprises
 (a) a heavy chain variable region (V H FAP) comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:9, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:10, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:11, and a light chain variable region (V L FAP) comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:12, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:13, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:14.   
     
     
         34 . The bispecific antigen binding molecule of  claim 32 , wherein the first subunit of the Fc domain comprises the amino acid substitutions K392D and K409D (numbering according to Kabat EU index) and the second subunit of the Fc region comprises the amino acid substitutions E356K and D399K (numbering according to Kabat EU index). 
     
     
         35 . The method of  claim 31 , wherein the bispecific antigen binding molecule is administered in combination with a chemotherapeutic agent, radiation therapy and/or other agents for use in cancer immunotherapy. 
     
     
         36 . The bispecific antigen binding molecule of  claim 14 , wherein the IgG Fc domain comprises an IgG1 Fc domain or an IgG4 Fc domain. 
     
     
         37 . The bispecific antigen binding molecule of  claim 15 , wherein the one or more amino acid substitution that reduces the binding affinity of the antigen binding molecule to an Fc receptor and/or effector function comprise particularly the amino acid mutations L234A, L235A and P329G (numbering according to Kabat EU index).