Anti-dr5 antibodies and methods of use thereof

Abstract

The present invention relates to monospecific or bispecific antibody molecules that specifically bind the human DR5 antigen. The invention relates in particular to DR5-specific antibody molecules of the IgG1 isotype having a mutation in the Fc region that enhances clustering of IgG molecules after cell-surface antigen binding leading to the induction of DR5 signalling, apoptosis and cell death. The invention further relates to a combination of antibody molecules binding different epitopes on DR5. The invention also relates to pharmaceutical compositions containing these molecules and the treatment of cancer using these compositions.

1 . An antibody comprising a Fc region of a human immunoglobulin IgG and an antigen binding region binding to human DR5, wherein the Fc region comprises a mutation of an amino acid position corresponding to E430, E345 or S440 in human IgG1, wherein the numbering is according to the EU Index. 
     
     
         2 - 4 . (canceled) 
     
     
         5 . The antibody according to  claim 1 , wherein the antigen binding region binds to an epitope on human DR5 comprising or requiring
 a) one or more amino acid residues located within amino acid residues 116-138 and one or more amino acid residues located within amino acid residues 139-166 of SEQ ID NO 46, or   b) one or more amino acid residues located within amino acid residues 79-138 of SEQ ID NO 46.   
     
     
         6 . (canceled) 
     
     
         7 . The antibody according to  claim 1 , wherein the antibody comprises a variable heavy chain region comprising CDR1, CDR2 and CDR3 domains and a variable light chain region comprising CDR1, CDR2 and CDR3 domains having the amino acid sequences selected from the group consisting of:
 a) SEQ ID NOs: 1, 2, 3 and SEQ ID NOs: 5, FAS, 6, respectively;   b) SEQ ID NOs: 1, 8, 3 and SEQ ID NOs: 5, FAS, 6, respectively;   c) SEQ ID NOs: 10, 2, 11 and SEQ ID NOs: 13, RTS, 14, respectively;   d) SEQ ID NOs: 16, 17, 18 and SEQ ID NOs: 21, GAS, 22, respectively; and   e) the heavy and light chain CDR1, CDR2, CDR3 sequences as defined in any one of a) to d) above having one to five mutations or substitutions in total across said CDR sequences.   
     
     
         8 . The antibody according to  claim 1 , wherein the antigen binding region comprises a variable heavy chain region and a variable light chain region having the amino acid sequences selected from the group consisting of:
 a) SEQ ID NO:4 and SEQ ID NO:7, respectively;   b) SEQ ID NO:9 and SEQ ID NO:7, respectively;   c) SEQ ID NO:12 and SEQ ID NO:15, respectively;   d) SEQ ID NO:19 and SEQ ID NO:23, respectively;   e) SEQ ID NO:20 and SEQ ID NO:23, respectively; and   f) the heavy and light chain variable region sequences as defined in any one of a) to e) above having one to five mutations or substitutions in total across said sequences.   
     
     
         9 . The antibody of  claim 1 , wherein the antibody is an IgG1, IgG2, IgG3, IgG4, IgE, IgD or IgM isotype. 
     
     
         10 - 11 . (canceled) 
     
     
         12 . The antibody according to  claim 1 , wherein the Fc region comprises an amino acid sequence selected from the group consisting of:
 a) SEQ ID NO:29;   b) SEQ ID NO:30;   c) SEQ ID NO:31;   d) SEQ ID NO:32; and   e) an amino acid sequence as defined in any one of a) to d) above having one to five mutations or substitutions in total across said sequence   
     
     
         13 . The antibody according to  claim 1 , comprising a heavy chain (HC) and a light chain (LC), wherein the LC comprises the sequence of SEQ ID NO:39 and wherein the HC comprises a sequence selected from the group consisting of:
 a) SEQ ID NO:33;   b) SEQ ID NO:34;   c) SEQ ID NO:35;   d) SEQ ID NO:36;   e) SEQ ID NO:37;   f) SEQ ID NO:38; and   g) the sequence as defined in any one of a) to f) above having one to five mutations or substitutions in total across said sequence.   
     
     
         14 . The antibody according to  claim 1 , comprising a heavy chain (HC) and a light chain (LC), wherein the LC comprises the sequence of SEQ ID NO:43 and wherein the HC comprises a sequence selected from the group consisting of:
 a) SEQ ID NO:40;   b) SEQ ID NO:41;   c) SEQ ID NO:42; or   d) the sequence as defined in any one of a) to c) above having one to five mutations or substitutions in total across said sequence.   
     
     
         15 . (canceled) 
     
     
         16 . The antibody according to  claim 1 , wherein the antibody is human, humanized or chimeric. 
     
     
         17 . The antibody according to  claim 1 , wherein the antibody is agonistic. 
     
     
         18 . The antibody according to  claim 1 , wherein the antibody
 (a) induces programmed cell death in a target cell, such as caspase dependent cell death:,   (b) induces apoptosis in a target cell expressing DR5; and/or   (c) reduces cell viability.   
     
     
         19 - 20 . (canceled) 
     
     
         21 . A composition comprising at least one antibody according to  claim 1  and a carrier. 
     
     
         22 . (canceled) 
     
     
         23 . The composition according to  claim 21 , which comprises a first antibody and a second antibody, wherein both the first antibody and second antibody comprise a Fc region of a human immunoglobulin IgG and an antigen binding region binding to human DR5, wherein the Fc region comprises a mutation of an amino acid position corresponding to E430, E345 or S440 in human IgG1. 
     
     
         24 - 31 . (canceled) 
     
     
         32 . A bispecific antibody comprising one or more antigen binding regions according to  claim 1 . 
     
     
         33 - 38 . (canceled) 
     
     
         39 . The bispecific antibody according to  claim 32 , wherein (a) the bispecific antibody comprises an Fc region comprising a first and a second heavy chain, said first heavy chain comprises a mutation corresponding to F405L in human IgG1 and said second heavy chain comprises a mutation corresponding to K409R in human IgG1, or (b) wherein the bispecific antibody comprises an Fc region comprising a first and a second heavy chain, said first heavy chain comprises a mutation corresponding to K409R in human IgG1 and said second heavy chain comprises a mutation corresponding to F405L in human IgG1. 
     
     
         40 - 43 . (canceled) 
     
     
         44 . A method of treating an infectious disease, autoimmune disease or cardiovascular anomalies comprising administering to a subject in need thereof an effective amount of the composition of  claim 21 . 
     
     
         45 . A method of treating a solid tumors tumor and/or hematological tumor comprising administering to a subject in need thereof an effective amount of the composition of  claim 21 . 
     
     
         46 . The method according to  claim 45 , wherein the solid tumor is selected from the group consisting of colorectal cancer, bladder cancer, osteosarcoma, chondrosarcoma, breast cancer, cancers of the central nervous system, cervical cancer, endometrium cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, sarcoma, and skin cancer; and the hematological tumor is selected from the group consisting of leukemia, lymphoma, and multiple myeloma. 
     
     
         47 - 48 . (canceled) 
     
     
         49 . A method for inducing apoptosis in DR5 expressing tumors comprising administering to a subject in need thereof an effective amount of the composition of  claim 21 . 
     
     
         50 . (canceled) 
     
     
         51 . A nucleic acid construct encoding an antibody according to  claim 1 . 
     
     
         52 . An expression vector comprising one or more nucleic acid constructs according to  claim 51 . 
     
     
         53 . A host cell comprising the expression vector of  claim 52 . 
     
     
         54 . (canceled) 
     
     
         55 . A method of treating cancer comprising administering to a subject in need thereof an effective amount of the antibody of  claim 1 . 
     
     
         56 . The method according to claim further comprising administering an additional therapeutic agent. 
     
     
         57 . The method according to  claim 56 , wherein the additional therapeutic agent is one or more anti-cancer agent(s) selected from the group consisting of:
 chemotherapeutics, kinase inhibitors, apoptosis-modulating agents, RAS inhibitors, proteasome inhibitors, histone deacetylase inhibitors, nutraceuticals, cytokines, antibodies or antibody mimetics, and antibody-drug conjugates.   
     
     
         58 . A kit comprising the antibody of  claim 1 , and instructions for use. 
     
     
         59 - 60 . (canceled)