ANTI-CD3 ANTIBODIES, BISPECIFIC ANTIGEN-BINDING MOLECULES THAT BIND CD3 AND CD20, AND USES THEREOF

• US 20140088295A1
• Filed: 09/19/2013
• Published: 03/27/2014
• Est. Priority Date: 09/21/2012
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: PAPADOPOULOS, Nicholas J., SMITH, Eric
• Granted Patent: US 9,657,102 B2
• Time in Patent Office: Days
• US Class Current: 530/387.3
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2317/74 Inducing cell proliferation
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides antibodies that bind to CD3 and methods of using the same. According to certain embodiments, the antibodies of the invention bind human CD3 with high affinity and induce human T cell proliferation. The invention includes antibodies that bind CD3 and induce T cell-mediated killing of tumor cells. According to certain embodiments, the present invention provides bispecific antigen-binding molecules comprising a first antigen-binding domain that specifically binds human CD3, and a second antigen-binding molecule that specifically binds human CD20. In certain embodiments, the bispecific antigen-binding molecules of the present invention are capable of inhibiting the growth of B-cell tumors expressing CD20. The antibodies and bispecific antigen-binding molecules of the invention are useful for the treatment of diseases and disorders in which an upregulated or induced targeted immune response is desired and/or therapeutically beneficial. For example, the antibodies of the invention are useful for the treatment of various cancers as well as other CD20-related diseases and disorders.

Claims

1-52. -52. (canceled)

53. A fully human bispecific antibody comprising a first antigen-binding domain that specifically binds human CD3, and a second antigen-binding domain that specifically binds human CD20.

54. The fully human bispecific antibody of claim 53, wherein the fully human bispecific antibody binds to CD3-expressing human T-cells with an EC₅₀ value of between 1×

10<sup>−

12</sup> M and 1×

10<sup>−

6</sup> M.

55. The fully human bispecific antibody of claim 54, wherein the fully human bispecific antibody binds to CD3-expressing human T-cells with an EC₅₀ value of between 1×

10<sup>−

9</sup> M and 6×

10<sup>−

8</sup> M.

56. The fully human bispecific antibody of claim 53, wherein the fully human bispecific antibody binds human cells expressing human CD3 and cynomolgus monkey cells expressing cynomolgus CD3.

57. The fully human bispecific antibody of claim 53, wherein the fully human bispecific antibody induces proliferation of human and cynomolgus peripheral blood mononuclear cells (PBMCs) in vitro.

58. The fully human bispecific antibody of claim 53, wherein the fully human bispecific antibody induces IFN-gamma release and CD25 up-regulation in human whole blood.

59. The fully human bispecific antibody of claim 53, wherein the fully human bispecific antibody induces T-cell mediated cytotoxicity of human B-cells.

60. The fully human bispecific antibody of claim 59, wherein the fully human bispecific antibody induces T-cell mediated cytotoxicity of human B-cells that are resistant to or refractory to anti-CD20-mediated cytotoxicity.

61. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain that specifically binds human CD3 comprises three heavy chain complementarity determining regions (A1-HCDR1, A1-HCDR2 and A1-HCDR3) from a heavy chain variable region (HCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

1250, 1266, 1282, 1298, 1314 and 1329, and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) from a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333.

62. The fully human bispecific antibody of claim 53, wherein the second antigen-binding domain that specifically binds human CD20 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) from a heavy chain variable region (HCVR) comprising SEQ ID NO:

1242, and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) from a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333.

63. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain that specifically binds human CD3 comprises three heavy chain complementarity determining regions (A1-HCDR1, A1-HCDR2 and A1-HCDR3) and three light chain complementarity determining regions (A1-LCDR1, A1-LCDR2 and A1-LCDR3), wherein A1-HCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:

1252, 1268, 1284, 1300, 1316 and 1330;

wherein A1-HCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1254, 1270, 1286, 1302, 1318 and 1331;

wherein A1-HCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1256, 1272, 1288, 1304, 1320 and 1332, wherein A1-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A1-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A1-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336.

64. The fully human bispecific antibody of claim 53, wherein the second antigen-binding domain that specifically binds human CD20 comprises three heavy chain complementarity determining regions (A2-HCDR1, A2-HCDR2 and A2-HCDR3) and three light chain complementarity determining regions (A2-LCDR1, A2-LCDR2 and A2-LCDR3), wherein A2-HCDR1 comprises the amino acid sequence of SEQ ID NO:

1244, wherein A2-HCDR2 comprises the amino acid sequence of SEQ ID NO;

1246, wherein A2-HCDR3 comprises SEQ ID NO;

1248, wherein A2-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A2-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A2-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336.

65. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain that specifically binds human CD3 comprises three heavy chain complementarity determining regions (A1-HCDR1, A1-HCDR2 and A1-HCDR3) and three light chain complementarity determining regions (A1-LCDR1, A1-LCDR2 and A1-LCDR3), and wherein the second antigen-binding domain that specifically binds human CD20 comprises three heavy chain complementarity determining regions (A2-HCDR1, A2-HCDR2 and A2-HCDR3) and three light chain complementarity determining regions (A2-LCDR1, A2-LCDR2 and A2-LCDR3);

wherein A1-HCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1252, 1268, 1284, 1300, 1316 and 1330;

wherein A1-HCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1254, 1270, 1286, 1302, 1318 and 1331;

wherein A1-HCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1256, 1272, 1288, 1304, 1320 and 1332, wherein A1-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A1-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A1-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336; and

wherein A2-HCDR1 comprises the amino acid sequence of SEQ ID NO;

1244, wherein A2-HCDR2 comprises the amino acid sequence of SEQ ID NO;

1246, wherein A2-HCDR3 comprises SEQ ID NO;

1248, wherein A2-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A2-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A2-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336.

66. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain competes for binding to human CD3 with a reference antigen-binding protein comprising three heavy chain complementarity determining regions (A1-HCDR1, A1-HCDR2 and A1-HCDR3) and three light chain complementarity determining regions (A1-LCDR1, A1-LCDR2 and A1-LCDR3), wherein A1-HCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:

1252, 1268, 1284, 1300, 1316 and 1330;

wherein A1-HCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1254, 1270, 1286, 1302, 1318 and 1331;

wherein A1-HCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1256, 1272, 1288, 1304, 1320 and 1332, wherein A1-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A1-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A1-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336.

67. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain competes for binding to human CD3 with a reference antigen-binding protein comprising a heavy chain variable region (HCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

1250, 1266, 1282, 1298, 1314 and 1329, and a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333.

68. The fully human bispecific antibody of claim 53, wherein the second antigen-binding domain competes for binding to human CD20 with a reference antigen-binding protein comprising three heavy chain complementarity determining regions (A2-HCDR1, A2-HCDR2 and A2-HCDR3) and three light chain complementarity determining regions (A2-LCDR1, A2-LCDR2 and A2-LCDR3), wherein A2-HCDR1 comprises the amino acid sequence of SEQ ID NO:

1244, wherein A2-HCDR2 comprises the amino acid sequence of SEQ ID NO;

1246, wherein A2-HCDR3 comprises SEQ ID NO;

1248, wherein A2-LCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1260, 1276, 1292, 1308, 1324 and 1334, wherein A2-LCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1262, 1278, 1294, 1310, 1326 and 1335, and wherein A2-LCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs;

1264, 1280, 1296, 1312, 1328 and 1336.

69. The fully human bispecific antibody of claim 53, wherein the second antigen-binding domain competes for binding to human CD20 with a reference antigen-binding protein comprising a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:

1242, and a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333.

70. The fully human bispecific antibody of claim 53, wherein the first antigen-binding domain competes for binding to human CD3 with a reference antigen-binding protein comprising a heavy chain variable region (HCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

1250, 1266, 1282, 1298, 1314 and 1329, and a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333; and

wherein the second antigen-binding domain competes for binding to human CD20 with a reference antigen-binding protein comprising a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO;

1242, and a light chain variable region (LCVR) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

1258, 1274, 1290, 1306, 1322 and 1333.

71. A fully human bispecific antibody comprising a first antigen-binding domain that specifically binds human CD3, and a second antigen-binding domain that specifically binds human CD20;

wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) comprising SEQ ID NO;

1250, and a light chain variable region (LCVR) comprising SEQ ID NO;

1258; and

wherein the second antigen-binding domain comprises an HCVR comprising SEQ ID NO;

1242, and a LCVR comprising SEQ ID NO;

1258.