Humanized anti-epiregulin antibody, and cancer therapeutic agent comprising said antibody as active ingredient

Patent metadata

Field	Value
Publication number	US-9556264-B2
Application number	US-201214369539-A
Country	US
Kind code	B2
Filing date	Dec 28, 2012
Priority date	Dec 28, 2011
Publication date	Jan 31, 2017
Grant date	Jan 31, 2017

Abstract

The inventors successfully produced anti-Epiregulin antibodies showing cross-species reactivity between cynomolgus monkey (non-human animals) and human, anti-Epiregulin antibodies with suppressed chemical degradation, anti-Epiregulin antibodies with lowered isoelectric point, anti-Epiregulin antibodies with increased thermal denaturation midpoint temperature, and anti-Epiregulin antibodies with reduced amount of aggregate by performing appropriate amino acid residue substitutions in the variable-region sequences of the humanized EP27 antibody which inhibits growth of cancer cells by exhibiting cytotoxic activity and neutralizing activity against human Epiregulin-expressing cancer cells.

First claim

The invention claimed is:

1. An anti-Epiregulin antibody which is an antibody that binds to an epitope bound by an anti-Epiregulin antibody comprising heavy-chain variable region CDRs of SEQ ID NOs: 9, 10, and 11 and light chain variable region CDRs of SEQ ID NO: 12, 13, and 14, wherein the antibody is characterized in having a smaller ratio of the KD value for monkey Epiregulin of SEQ ID NO: 170 (cEREG KD) to the KD value for human Epiregulin of SEQ ID NO: 34 (hEREG KD) (cEREG KD/hEREG KD) than the cEREG KD/hEREG KD ratio of the anti-Epiregulin antibody comprising heavy-chain variable region CDRs of SEQ ID NOs: 9, 10, and 11 and light-chain variable region CDRs of SEQ ID NO: 12, 13, and 14, which is selected from any one of (1) to (8) below:
(1) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR 3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light- chain CDR 3 of SEQ ID NO: 14;
(2) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR 3 of SEQ ID NO: 152; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 14;
(3) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 14;
(4) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 160, and a heavy-chain CDR3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 14;
(5) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 164;
(6) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR3 of SEQ ID NO: 152; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 164;
(7) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 153, and a heavy-chain CDR3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 164; and
(8) an anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 160, and a heavy-chain CDR3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR3 of SEQ ID NO: 164.

2. The antibody of claim 1 , which comprises the heavy-chain constant region of SEQ ID NO: 26.

3. The antibody of claim 2 , wherein the heavy-chain constant region of SEQ ID NO: 26 comprises at least one substitution of amino acid at a position selected from the group consisting of 230, 240, 244, 245, 247, 262, 263, 266, 273, 275, 299, 302, 313, 323, 325, 328, and 332 as indicated by EU numbering.

4. The antibody of claim 1 , which comprises the light-chain constant region of SEQ ID NO: 27.

5. The antibody of claim 1 , which has a neutralizing activity.

6. The antibody of claim 1 , which has cytotoxicity.

7. The antibody of claim 6 , wherein the cytotoxicity is CDC and/or ADCC.

8. The antibody of claim 1 , wherein a growth inhibitor or a cytotoxic substance is linked to the antibody.

9. A pharmaceutical composition comprising the antibody of claim 1 as an active ingredient.

10. An anti-Epiregulin antibody which is an antibody that binds to an epitope bound by an anti-Epiregulin antibody comprising heavy-chain variable region CDRs of SEQ ID NOs: 9, 10, and 11 and light chain variable region CDRs of SEQ ID NO: 12, 13, and 14, wherein the antibody is characterized in having a smaller ratio of the KD value for monkey Epiregulin of SEQ ID NO: 170 (cEREG KD) to the KD value for human Epiregulin of SEQ ID NO: 34 (hEREG KD) (cEREG KD/hEREG KD) than the cEREG KD/hEREG KD ratio of the anti-Epiregulin antibody comprising heavy-chain variable region CDRs of SEQ ID NOs: 9, 10, and 11 and light-chain variable region CDRs of SEQ ID NO: 12, 13, and 14, which is selected from any one of (1) to (8) below:
(1) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 140 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 136;
(2) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 142 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 136;
(3) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 149 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 136;
(4) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 150 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 136;
(5) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 140 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 141;
(6) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 142 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 141;
(7) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: t49 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 141; and
(8) an anti-Epiregulin antibody which comprises a heavy-chain variable region corresponding to the heavy-chain variable region in SEQ ID NO: 150 and a light-chain variable region corresponding to the light-chain variable region in SEQ ID NO: 141.

11. An anti-Epiregulin antibody which comprises a heavy-chain variable region comprising a heavy-chain CDR1 of SEQ ID NO: 9, a heavy-chain CDR2 of SEQ ID NO: 160, and a heavy-chain CDR 3 of SEQ ID NO: 158; and a light-chain variable region comprising a light-chain CDR1 of SEQ ID NO: 163, a light-chain CDR2 of SEQ ID NO: 13, and a light-chain CDR 3 of SEQ ID NO: 164.

12. The antibody of claim 11 , which comprises the heavy-chain co

Assignees

• Chugai Pharmaceutical Co Ltd

Inventors

• Shiraiwa Hirotake
• Esaki Keiko
• Igawa Tomoyuki
• Kuramochi Taichi
• Maeda Atsuhiko
• Tamba Shigero
• Tsunoda Hiroyuki
• Tachibana Tatsuhiko
• Kinoshita Yasuko
• Suzuki Masami
• Kato Atsuhiko
• Takeiri Etsuko
• Hashimoto Eri
• Watanabe Yoshinori

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P35/04

  specific for metastasis · CPC title

• C07K2317/77

  Internalization into the cell · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/734

  Complement-dependent cytotoxicity [CDC] · CPC title