MUTATED ANTI-TNFa ANTIBODIES AND METHODS OF THEIR USE

Abstract

The present invention is directed to modified antibodies, including anti-TNFα antibodies, in which C-terminal amino acids of heavy chain sequences are modified from a native sequence of proline-glycine-lysine (“PGK”) to one that includes a proline positioned between the glycine and lysine, resulting in a C-terminal sequence of proline-glycine-proline-lycine (“PGPK”). The invention further provides methods of producing and using such antibodies.

1 . An antibody, or antigen-binding portion thereof, comprising a C-terminal heavy chain sequence of proline-glycine-proline-lysine (PGPK) (SEQ ID NO:9). 
     
     
         2 . The antibody, or antigen-binding portion thereof, of  claim 1 , wherein the antibody, or antigen-binding portion thereof, is resistant to C-terminal processing by a carboxypeptidase. 
     
     
         3 . The antibody, or antigen-binding portion thereof, of  claim 1 , wherein the antibody, or antigen-binding portion thereof, is an anti-TNFα antibody, or antigen-binding portion thereof. 
     
     
         4 . The antibody, or antigen-binding portion thereof, of  claim 1 , wherein the antibody, or antigen-binding portion thereof, exhibits increased tissue penetration, increased TNFα affinity, reduced tissue destruction, reduced bone erosion, reduced synovial proliferation, reduced cell infiltration, reduced chondrocyte death, reduced proteoglycan loss, increased protection against the development of arthritic scores when administered to an animal model of arthritis, or increased protection against the development of histopathology scores when administered to an animal model of arthritis as compared to an antibody, or antigen-binding portion thereof, comprising a C-terminal heavy chain sequence of proline-glycine-lysine (PGK) (SEQ ID NO:10). 
     
     
         5 . A composition comprising an antibody, or antigen-binding portion thereof, wherein the composition comprises less than about 50% lysine variant species that lack a C-terminal lysine (Lys 0). 
     
     
         6 . The composition of  claim 5 , wherein the composition comprises less than about 25% lysine variant species that have one C-terminal lysine (Lys 1). 
     
     
         7 . The composition of  claim 5 , wherein the composition comprises at least about 70% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         8 . The composition of  claim 7 , wherein the composition comprises at least about 80% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         9 . The composition of  claim 8 , wherein the composition comprises at least about 90% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         10 . The composition of  claim 9 , wherein the composition comprises at least about 95% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         11 . The composition of any  claim 5 , wherein the composition comprises less than about 10% acidic species, wherein the acidic species comprise a first acidic species region (AR1) and a second acidic species region (AR2). 
     
     
         12 . The composition of  claim 11 , wherein the composition comprises about 0% AR1. 
     
     
         13 . The composition of  claim 11 , wherein the composition comprises less than about 4% AR2. 
     
     
         14 . The composition of  claim 11 , wherein the composition comprises about 0% AR1 and about 3% AR2. 
     
     
         15 . A composition comprising an antibody, or antigen-binding portion thereof, wherein the composition comprises at least about 70% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         16 . The composition of  claim 15 , wherein the composition comprises at least about 80% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         17 . The composition of  claim 15 , wherein the composition comprises at least about 90% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         18 . The composition of  claim 15 , wherein the composition comprises at least about 100% lysine variant species that have two C-terminal lysines (Lys 2). 
     
     
         19 . The composition of any  claim 15 , wherein the composition comprises less than about 10% acidic species, wherein the acidic species comprise a first acidic species region (AR1) and a second acidic species region (AR2). 
     
     
         20 . The composition of  claim 19 , wherein the composition comprises about 0% AR1. 
     
     
         21 . The composition of  claim 19 , wherein the composition comprises less than about 4% AR2. 
     
     
         22 . The composition of  claim 19 , wherein the composition comprises about 0% ARI and about 3% AR2. 
     
     
         23 . The composition of  claim 15 , wherein the antibody, or antigen-binding portion thereof, is an anti-TNFα antibody, or antigen-binding portion thereof. 
     
     
         24 . The composition of  claim 23 , wherein the antibody, or antigen-binding portion thereof, comprises a light chain variable region (LCVR) having a CDR1 domain comprising the amino acid sequence of SEQ ID NO:7, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO:3; and a heavy chain variable region (HCVR) having a CDR1 domain comprising the amino acid sequence of SEQ ID NO:8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:6, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO:4. 
     
     
         25 . The composition of  claim 24 , wherein the antibody, or antigen-binding portion thereof, comprises a LCVR comprising the amino acid sequence set forth in SEQ ID NO:1 and a HCVR comprising the amino acid sequence set forth in SEQ ID NO:2. 
     
     
         26 . The composition of  claim 25 , wherein the antibody, or antigen-binding portion thereof, comprises adalimumab, or an antigen-binding portion thereof. 
     
     
         27 . The composition of  claim 15 , wherein the antibody, or antigen-binding portion thereof, is resistant to C-terminal processing by a carboxypeptidase. 
     
     
         28 . The composition of  claim 15 , wherein the antibody, or antigen-binding portion thereof, exhibits increased tissue penetration, increased TNFα affinity, reduced tissue destruction, reduced bone erosion, reduced synovial proliferation, reduced cell infiltration, reduced chondrocyte death, reduced proteoglycan loss, increased protection against the development of arthritic scores when administered to an animal model of arthritis, or increased protection against the development of histopathology scores when administered to an animal model of arthritis as compared to an antibody, or antigen-binding portion thereof, comprising a C-terminal heavy chain sequence of proline-glycine-lysine (PGK) (SEQ ID NO:10). 
     
     
         29 . A method of treating a subject having a disorder in which TNFα activity is detrimental, the method comprising administering a therapeutically effective amount of the antibody, or antigen-binding portion thereof, of  claim 1  to the subject, thereby treating the TNFα-associated disease or disorder. 
     
     
         30 . The method of  claim 29 , wherein the disorder in which TNFα activity is detrimental is selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's Disease plaque psoriasis, active axial spondyloarthritis and non-radiographic axial spondyloarthritis.