Humanized anti-cd19 antibodies and their use in treatment of oncology, transplantation and autoimmune disease

Abstract

The present invention provides chimeric and humanized versions of anti-CD19 mouse monoclonal antibodies. The invention further relates to pharmaceutical compositions, immunotherapeutic compositions, and methods using therapeutic antibodies that bind to the human CD19 antigen and that may mediate ADCC, CDC, and/or apoptosis for the treatment of B cell diseases and disorders, such as, but not limited to, B cell malignancies, for the treatment and prevention of autoimmune disease, and for the treatment and prevention of graft-versus-host disease (GVHD), humoral rejection, and post-transplantation lymphoproliferative disorder in human transplant recipients.

What is claimed is: 
     
         1 . An isolated purified chimeric, humanized or human monoclonal antibody or fragment thereof comprising a VH comprising the amino acid sequence of SEQ ID NO.:237 or a VL comprising the amino acid sequence of SEQ ID NO.:238, wherein said antibody binds a human CD19 antigen. 
     
     
         2 . The antibody of  claim 1 , wherein said antibody comprises a VH comprising the amino acid sequence of SEQ ID NO.:237 and a VL comprising the amino acid sequence of SEQ ID NO.:238. 
     
     
         3 . The antibody of  claim 1 , wherein said VH comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 34, 44, 102, 103, 104, 105, 106, 107, 108, 109, 191, 192, and 236; and wherein said VK comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 52, 62, 68, 70, 74, 76, 78, 80, 110, 111, 193, 194, 195, 196, 917, 198, 199, 200, 201, 202, 203, 204, 205, 206, and 207. 
     
     
         4 . The antibody of  claim 2 , wherein said VH comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 34, 44, 102, 103, 104, 105, 106, 107, 108, 109, 191, 192, and 236; and wherein said VK comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 52, 62, 68, 70, 74, 76, 78, 80, 110, 111, 193, 194, 195, 196, 917, 198, 199, 200, 201, 202, 203, 204, 205, 206, and 207. 
     
     
         5 . The antibody of  claim 4 , wherein said antibody is selected from the group consisting of:
 a) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:34 and VL comprising the amino acid sequence of SEQ ID NO.: 68,   b) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:102 and VL comprising the amino acid sequence of SEQ ID NO.:110,   c) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:103 and VL comprising the amino acid sequence of SEQ ID NO.:111,   d) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:103 and VL comprising the amino acid sequence of SEQ ID NO.:113,   e) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:104 and VL comprising the amino acid sequence of SEQ ID NO.:112,   f) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:105 and VL comprising the amino acid sequence of SEQ ID NO.:111,   g) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:111,   h) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:107 and VL comprising the amino acid sequence of SEQ ID NO.:111,   i) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:108 and VL comprising the amino acid sequence of SEQ ID NO.:111,   j) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:109 and VL comprising the amino acid sequence of SEQ ID NO.:111,   k) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:111,   l) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:193,   m) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:194,   n) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:195,   o) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:192 and VL comprising the amino acid sequence of SEQ ID NO.:196,   p) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:197,   q) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:198,   r) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:199,   s) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:200,   t) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:201,   u) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:202,   v) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:203,   w) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:198,   x) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:192 and VL comprising the amino acid sequence of SEQ ID NO.:204,   y) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:205,   z) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:206,   aa) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:197, and   bb) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:207.   
     
     
         6 . The antibody of  claim 4 , wherein said antibody possesses a functional activity selected from the group consisting of: enhanced ADCC activity, induction of B cell apoptosis, inhibition of B cell proliferation, and in vivo B cell depletion. 
     
     
         7 . The antibody of  claim 4 , wherein said antibody has complex N-glycoside-linked sugar chains bound to the Fc region in which fucose is not bound to N-acetylglucosamine in the reducing end in the sugar chain. 
     
     
         8 . The antibody of  claim 7 , wherein said antibody efficiently mediates in vitro ADCC activity against Karpas-422, Karpas-1106P, and DB cell lines but not against Granta-519 cell line. 
     
     
         9 . The antibody of  claim 7 , wherein said antibody is capable of depleting B cells in an animal model, wherein said B cells are selected from a group consisting of: circulating B cells, blood B cells, splenic B cells, marginal zone B cells, follicular B cells, peritoneal B cells, and/or bone marrow B cells. 
     
     
         10 . The antibody of  claim 9 , wherein said depletion reduces B cell levels by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% seven days after administration of 2.5 mg/kg dose of said antibody. 
     
     
         11 . The antibody of  claim 4 , wherein said antibody is an Fc variant, wherein said variant Fc has an altered affinity for one or more Fc ligand selected from the group consisting of: Clq, FcγRI, FcγRIIA, FcγRIIB, FcγRIIIA and FcγRIV. 
     
     
         12 . The antibody of  claim 11 , wherein said Fc variant has an affinity for the Fc receptor FcγRIIIA that is at least about 5 fold lower than that of a comparable molecule, and wherein said Fc variant has an affinity for the Fc receptor FcγRI1B that is within about 2 fold of that of a comparable molecule. 
     
     
         13 . The antibody of  claim 12 , wherein said Fc variant comprises mutations that result in enhanced ADCC activity. 
     
     
         14 . An isolated nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 34, 44, 102, 103, 104, 105, 106, 107, 108, 109, 191, 192, 236, 52, 62, 68, 70, 74, 76, 78, 80, 110, 111, 193, 194, 195, 196, 917, 198, 199, 200, 201, 202, 203, 204, 205, 206, and 207. 
     
     
         15 . An isolated cell expressing the antibody of  claim 4 . 
     
     
         16 . A pharmaceutical composition comprising the antibody of  claim 4  in a pharmaceutically-acceptable carrier. 
     
     
         17 . A method of treating a B cell disease or disorder in a human comprising: administering to a human in need thereof a therapeutically-effective amount of an antibody,
 a) wherein said antibody is a chimeric, humanized or human monoclonal antibody or fragment thereof,   b) wherein said antibody comprises a VH or a VL wherein said VH comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 34, 44, 102, 103, 104, 105, 106, 107, 108, 109, 191, 192, and 236,   c) wherein said VL comprises an amino acid sequence selected from the group consisting of: SEQ ID NO.: 52, 62, 68, 70, 74, 76, 78, 80, 110, 111, 193, 194, 195, 196, 917, 198, 199, 200, 201, 202, 203, 204, 205, 206, and 207,   d) wherein said antibody binds a human CD19 antigen, and   e) wherein said disease or disorder is selected from a group consisting of: B cell malignancy, autoimmune disease, autoimmune disorder, humoral rejection in a human transplant patient, graft-versus-host disease (GVHD) and post-transplantation lymphoproliferative disorder in human transplant recipient.   
     
     
         18 . The method of  claim 17 , wherein said antibody is selected from the group consisting of:
 a) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:34 and VL comprising the amino acid sequence of SEQ ID NO.: 68,   b) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:102 and VL comprising the amino acid sequence of SEQ ID NO.:110,   c) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:103 and VL comprising the amino acid sequence of SEQ ID NO.:111,   d) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:103 and VL comprising the amino acid sequence of SEQ ID NO.:113,   e) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:104 and VL comprising the amino acid sequence of SEQ ID NO.:112,   f) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:105 and VL comprising the amino acid sequence of SEQ ID NO.:111,   g) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:111,   h) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:107 and VL comprising the amino acid sequence of SEQ TD NO.:111,   i) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:108 and VL comprising the amino acid sequence of SEQ ID NO.:111,   j) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:109 and VL comprising the amino acid sequence of SEQ ID NO.:111,   k) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:111,   l) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:193,   m) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:194,   n) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:195,   o) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:192 and VL comprising the amino acid sequence of SEQ ID NO.:196,   p) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:197,   q) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:198,   r) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:199,   s) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:200,   t) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:201,   u) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:202,   v) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:203,   w) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:106 and VL comprising the amino acid sequence of SEQ ID NO.:198,   x) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:192 and VL comprising the amino acid sequence of SEQ TD NO.:204,   y) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:205,   z) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:206,   aa) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:197, and   bb) an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO.:191 and VL comprising the amino acid sequence of SEQ ID NO.:207.   
     
     
         19 . The method of claimed  17 , wherein said method comprises the depletion of B cells selected from the group consisting of: circulating B cells, blood B cells, splenic B cells, marginal zone B cells, follicular B cells, peritoneal B cells, and/or bone marrow B cells. 
     
     
         20 . The method of claimed  17 , wherein said method comprises the depletion of B cells selected from the group consisting of: progenitor B cells, early pro-B cells, late pro-B cells, large-pre-B cells, small pre-B cells, immature B cells, mature B cells, antigen stimulated B cells, and/or plasma cells. 
     
     
         21 . The method of  claim 17 , wherein said antibody has an enhanced ADCC activity. 
     
     
         22 . The method of  claim 21 , wherein said depletion reduces B cell levels by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100%. 
     
     
         23 . The method of  claim 21 , wherein said depletion persist for a time period selected from the group consisting of: at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months or at least 12 months.