High affinity antibodies to human IL-6 receptor

• US 20070280945A1
• Filed: 06/01/2007
• Published: 12/06/2007
• Est. Priority Date: 06/02/2006
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Stevens, Sean, Rafique, Ashique, Smith, Eric, Chen, Gang, Karow, Margaret, Fandl, James, Papadopoulos, Nicholas, Pobursky, Kevin, Fairhurst, Jeanette, Martin, Joel, Huang, Tammy
• Granted Patent: US 7,582,298 B2
• Time in Patent Office: Days
• US Class Current: 424/145.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

A human antibody or an antigen-binding fragment which binds human IL-6 receptor (hIL-6R) with a K_D of about 500 pM or less and blocks IL-6 activity with an IC₅₀ of 200 pM or less. In preferred embodiments, the antibody the antibody or antigen-binding fragment binds hIL-6R with an affinity at least 2-fold higher relative to its binding monkey IL-6R.

Claims

1. An antibody or antigen-binding fragment thereof, which specifically binds human interleukin-6 receptor (hIL-6R) with a K_D of about 500 pM or less, as measured by surface plasmon resonance.

2. The antibody or antigen-binding fragment of claim 1, wherein the K_D is less than 300 pM.

3. The antibody or antigen-binding fragment of claim 1 which binds hIL-6R with an affinity at least 2-fold higher relative to binding monkey IL-6R.

4. The antibody or antigen-binding fragment of claim 1, comprising a heavy chain variable region (HCVR) selected from the group consisting of SEQ ID NO:

3, 227, 19, 231, 35, 51, 67, 83, 99, 115, 131, 147, 239, 241, 163, 179, 195 and 211.

5. The antibody or antigen-binding fragment of claim 4, further comprising a light chain variable region (LCVR) selected from the group consisting of SEQ ID NO:

11, 229, 27, 233, 43, 59, 75, 91, 107, 123, 139, 155, 171, 187, 237, 203 and 219.

6. The antibody or antigen-binding fragment of claim 1, comprising HCVR/LCVR pairs selected from the group consisting of SEQ ID NO:

3/11;

227/229;

19/27;

231/233;

35/43;

51/59;

67/75;

83/91;

99/107;

115/123;

131/139;

147/155;

239/155;

241;

155;

163/171;

179/187;

235/237;

195/203 and 211/219.

11. The antibody or antigen-binding fragment of claim 1, comprising:

a heavy chain CDR3 domain selected from the group consisting of SEQ ID NO;

25, 153, 9, 185, 41, 57, 73, 89, 105, 121, 137, 169, 201 and 217; and

a light chain CDR3 domain selected from the group consisting of SEQ ID NO;

33, 161, 17, 193, 49, 65, 81, 97, 113, 129, 145, 177, 209 and 225.

12. The antibody or antigen-binding fragment of claim 11, further comprises:

a heavy chain CDR1 domain selected from the group consisting of SEQ ID NO;

21, 149, 5, 181, 37, 53, 69, 85, 101, 117, 133, 165, 197, and 213;

a heavy chain CDR2 domain selected from the group consisting of SEQ ID NO;

23, 151, 7, 183, 39, 55, 71, 87, 103, 119, 135, 167, 199 and 215;

a light chain CDR1 domain selected from the group consisting of SEQ ID NO;

29, 157, 13, 189, 45, 61, 77, 93, 109, 125, 141, 173, 205 and 221; and

a light chain CDR2 domain selected from the group consisting of SEQ ID NO;

31, 159, 15, 191, 47, 63, 79, 95, 111, 127, 143, 175, 207 and 223.

13. An isolated nucleic acid molecule encoding the antibody or antigen-binding fragment of claim 11, wherein

a heavy chain CDR3 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:

24, 152, 8, 184, 40, 56, 72, 88, 104, 120, 136, 168, 200 and 216; and

a light chain CDR3 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO;

32, 160, 16, 192, 48, 64, 80, 96, 112, 128, 144, 176, 208 and 224.

14. The isolated nucleic acid molecule of claim 12, wherein

a heavy chain CDR1 encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:

20, 148, 4, 180, 36, 52, 68, 84, 100, 116, 132, 164, 196 and 212;

a heavy chain CDR2 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO;

22, 150, 6, 182, 38, 54, 70, 86, 102, 118, 134, 166, 198 and 214;

a light chain CDR1 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO;

28, 156, 12, 188, 44, 60, 76, 92, 108, 124, 140, 172, 204 and 220; and

a light chain CDR2 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO;

30, 158, 14, 190, 46, 62, 78, 94, 110, 126, 142, 174, 206 and 222.

19. A pharmaceutical composition comprising the antibody or antigen-binding fragment of claim 1 and a pharmaceutically acceptable carrier.

20. A method for treating an interleukin-6 (IL-6) mediated disease or disorder, comprising administering a pharmaceutical effective amount of the antibody of claim 1 to a subject in need thereof.

7. An isolated nucleic acid molecule encoding an antibody or antigen-binding fragment thereof which specifically binds human interleukin-6 receptor (hIL-6R), comprising a heavy chain variable region (HCVR) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:

2, 226, 18, 230, 34, 50, 66, 82, 98, 114, 130, 146, 238, 240, 162, 178, 234, 194 and 210.

8. The nucleic acid molecule of claim 7, wherein the antibody or antigen-binding fragment further comprises a light chain variable region (LCVR) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:

10, 228, 26, 232, 42, 58, 74, 90, 106, 122, 138, 154, 170, 186, 236, 202 and 218.

15. A vector comprising the nucleotide sequence of claim 7.

9. An antibody or antigen-binding fragment which specifically binds human interleukin-6 receptor (hIL-6R), comprising a heavy chain complementary determining region 3 (CDR3) and a light chain CDR3, wherein

the heavy chain CDR3 domain comprises an amino acid sequence of the formula X¹-X²-X³-X⁴-X⁵-X⁶-X⁷-X⁸-X⁹-X¹⁰-X¹¹-X¹²-X¹³-X¹⁴-X¹⁵-X¹⁶-X¹⁷-X¹⁸-X¹⁹ (SEQ ID NO:

247) wherein X¹=Ala, X²=Lys, X³=Gly, X⁴=Arg, X⁵=Asp, X⁶=Ser or Ala, X⁷=Phe, X⁸=Asp;

X⁹=Ile, X¹⁰=Pro or absent, X¹¹=Phe or absent, X¹²=Val or absent, X¹³=Tyr or absent, X¹⁴=Tyr or absent, X¹⁵=Tyr or absent, X¹⁶=Gly or absent, X¹⁷=Met or absent, X¹⁸=Asp or absent, and X¹⁹=Val or absent; and

the light chain CDR3 domain comprises an amino acid sequence of the formula X¹-X²-X³-X⁴-X⁵-X⁶-X⁷-X⁸-X⁹ (SEQ ID NO;

250) wherein X¹=Gln, X²=Gln or His, X³=Ala, X⁴=Asn or Tyr, X⁵=Ser, X⁶=Phe, X⁷=Pro, X⁸=Pro, and X⁹=Thr.

10. The antibody or antigen-binding fragment of claim 9, further comprising

a heavy chain CDR1 domain comprising an amino acid sequence of the formula X¹-X²-X³-X⁴-X⁵-X⁶-X⁷-X⁸ (SEQ ID NO:

245) wherein X¹=Gly or Arg, X²=Phe, X³=Thr, X⁴=Phe, X⁵=Asp, X⁶=Asp, X⁷=Tyr, and X⁸=Ala;

a heavy chain CDR2 domain comprising an amino acid sequence of the formula X¹-X²-X³-X⁴-X⁵-X⁶-X⁷-X⁸ (SEQ ID NO;

246) wherein X¹=Ile or Val, X²=Ser, X³=Trp, X⁴=Asn, X⁵=Ser, X⁶=Gly, X⁷=Ser, and X⁸=lie;

light chain CDR1 domain comprising an amino acid sequence of the formula X¹-X²-X³-X⁴-X⁵-X⁶-X⁷-X⁸ (SEQ ID NO;

248), wherein X¹=Gln, X²=Gly, X³=lie, X⁴=Ser, X⁵=Ser, and X⁶=Trp; and

a light chain CDR2 domain comprising an amino acid sequence of the formula X¹-X²-X³ (SEQ ID NO;

249), wherein X¹=Gly or Ala, X²=Ala, and X³=Ser.

16. A host-vector system for the production of an antibody or antigen-binding fragment of an antibody which specifically binds human IL-6 receptor, comprising the vector of claim 16 in a suitable host cell.

17. The host-vector system of claim 16, wherein the host cell is a prokaryotic or eukaryotic cell selected from one of an E. coli or a CHO cell.

18. A method for producing an anti-IL-6R antibody or antigen-binding fragment thereof, comprising growing cells of the host-vector system of claim 16 under conditions permitting production of the antibody or fragment thereof and recovering the antibody or fragment so produced.