Modulated lysine variant species compositions and methods for producing and using the same

Abstract

The instant invention relates to modulated lysine variant species compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, and methods, e.g., cell culture and/or protein purification methods, for producing such modulated lysine variant species compositions. Methods for using such compositions to treat a disorder, e.g., a disorder in which TNFα is detrimental, are also provided.

1 . A method for treating a subject having a disorder in which TNFα activity is detrimental, the method comprising administering a therapeutically effective amount of a composition comprising a human anti-TNFα antibody, wherein less than 65% of the lysine variant species in said composition have zero C-terminal lysines (Lys 0),
 wherein the lysine variant species include the main peak and peaks that elute at a relative residence time later than the main peak, as detected using weak cation-exchange chromatography, and 
 wherein the human anti-TNFα antibody 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; 
 thereby treating the TNFα-associated disease or disorder. 
 
     
     
         2 . The method of  claim 1 , wherein the disorder in which TNFα activity is detrimental is selected from the group consisting of rheumatoid arthritis (RA), juvenile idiopathic arthritic, psoriatic arthritis, ankylosing spondylitis, Crohn's Disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa (HS), uveitis, active axial spondyloarthritis (active axSpA) and non-radiographic axial spondyloarthritis (nr-axSpA). 
     
     
         3 . The method of  claim 1 , wherein said human anti-TNFα antibody is adalimumab. 
     
     
         4 . The method of  claim 3 , wherein less than 60% of the lysine variant species in said composition have zero C-terminal lysines (Lys 0). 
     
     
         5 . The method of  claim 3 , wherein 50-60% of the lysine variant species in said composition have zero C-terminal lysines (Lys 0). 
     
     
         6 . The method of  claim 3 , wherein less than 55% of the lysine variant species in said composition have zero C-terminal lysines (Lys 0). 
     
     
         7 . The method of  claim 1 , wherein said composition is lyophilized. 
     
     
         8 . The method of  claim 3 , wherein said adalimumab is produced in a mammalian host cell grown in cell culture. 
     
     
         9 . The method of  claim 8 , wherein the mammalian host cell is selected from the group consisting of a CHO cell, an NSO cell, a COS cell, and an SP2 cell. 
     
     
         10 . The method of  claim 1 , wherein the composition is a pharmaceutical composition further comprising pharmaceutically acceptable carrier. 
     
     
         11 . The method of  claim 10 , wherein adalimumab is present in said pharmaceutical composition at a concentration of 25-100 mg/ml. 
     
     
         12 . The method of  claim 10 , wherein said pharmaceutical composition comprises one or more excipient selected from the group consisting of a buffering agent, a surfactant and a polyol, or a combination thereof. 
     
     
         13 . A method for treating a subject having a disorder in which TNFα activity is detrimental, the method comprising administering a therapeutically effective amount of a composition comprising a human anti-TNFα antibody, wherein the sum of the lysine variant species having one C-terminal lysine (Lys 1) and the lysine variant species having two C-terminal lysines (Lys 2) in said composition is greater than 35%,
 wherein the lysine variant species include the main peak and peaks that elute at a relative residence time later than the main peak, as detected using weak cation-exchange chromatography, 
 and wherein the human anti-TNFα antibody 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; 
 thereby treating the TNFα-associated disease or disorder. 
 
     
     
         14 . The method of  claim 13 , wherein the disorder in which TNFα activity is detrimental is selected from the group consisting of rheumatoid arthritis (RA), juvenile idiopathic arthritic, psoriatic arthritis, ankylosing spondylitis, Crohn's Disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa (HS), uveitis, active axial spondyloarthritis (active axSpA) and non-radiographic axial spondyloarthritis (nr-axSpA). 
     
     
         15 . The method of  claim 13 , wherein said human anti-TNFα antibody is adalimumab. 
     
     
         16 . The method of  claim 15 , wherein the sum of the lysine variant species having one C-terminal lysine (Lys 1) and the lysine variant species having two C-terminal lysines (Lys 2) in said composition is greater than 40%. 
     
     
         17 . The method of  claim 15 , wherein the sum of the lysine variant species having one C-terminal lysine (Lys 1) and the lysine variant species having two C-terminal lysines (Lys 2) in said composition is 40-50%. 
     
     
         18 . The method of  claim 15 , wherein the sum of the lysine variant species having one C-terminal lysine (Lys 1) and the lysine variant species having two C-terminal lysines (Lys 2) in said composition is greater than 50%. 
     
     
         19 . The method of  claim 15 , wherein greater than 25% of the lysine variant species in said composition have one C-terminal lysine (Lys 1). 
     
     
         20 . The method of  claim 15 , wherein greater than 30% of the lysine variant species in said composition have one C-terminal lysine (Lys 1). 
     
     
         21 . The method of  claim 13 , wherein said composition is lyophilized. 
     
     
         22 . The method of  claim 15 , wherein said adalimumab is produced in a mammalian host cell grown in cell culture. 
     
     
         23 . The method of  claim 22 , wherein the mammalian host cell is selected from the group consisting of a CHO cell, an NSO cell, a COS cell, and an SP2 cell. 
     
     
         24 . The method of  claim 13 , wherein the composition is a pharmaceutical composition further comprising pharmaceutically acceptable carrier. 
     
     
         25 . The method of  claim 24 , wherein adalimumab is present in said pharmaceutical composition at a concentration of 25-100 mg/ml. 
     
     
         26 . The method of  claim 24 , wherein said pharmaceutical composition comprises one or more excipient selected from the group consisting of a buffering agent, a surfactant and a polyol, or a combination thereof.