Cytotoxicity-inducing therapeutic agent

A multispecific antigen-binding molecule that comprises:

• (1) a domain comprising an antibody variable region having glypican 3-binding activity,
• (2) a domain comprising an antibody variable region having T-cell receptor complex-binding activity, and
• (3) a domain comprising an Fc region with reduced binding activity towards an Fcγ receptor,
• wherein the antibody variable region of (1) and the antibody variable region of (2) have a common light chain amino acid sequence;
• and wherein the multispecific antigen-binding molecule has a cytotoxic activity equivalent to or greater than the cytotoxic activity of a bispecific antibody having a glypican 3-binding domain comprising SEQ ID NOs:
• 47 and 48, and a T-cell receptor complex-binding domain comprising SEQ ID NOs:
• 49 and 50.

A bispecific antibody comprising a first and a second antibody variable domain selected from the group consisting of:

• (h1) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:424 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h2) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h3) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:299;
• (h4) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:301;
• (h5) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:302;
• (h6) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:304;
• (h7) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:306;
• (h8) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:307;
• (h9) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61n;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:103 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:309;
• (h10) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:122 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h11) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:129 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h12) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:215 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:132 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h13) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:197 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:128 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:299;
• (h14) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:197 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:128 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:310;
• (h15) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:197 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:128 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:319;
• (h16) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:197 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:128 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:53;
• (h17) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:211 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:142 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:299;
• (h18) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:211 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:142 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• (h19) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:211 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:144 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• (h20) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:206 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:144 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• (h21) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:206 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:142 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• (h22) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:206 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:164 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• (h24) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:211 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:164 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223;
• and
• (h25) a first antibody variable domain having glypican 3-binding activity that comprises an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:211 and a constant region having the amino acid sequence of SEQ ID NO:61;
• a second antibody variable domain having T-cell receptor complex-binding activity that comprises an heavy chain variable region having the amino acid sequence of SEQ ID NO:168 and a constant region having the amino acid sequence of SEQ ID NO:60 or 62;
• and a common antibody light chain variable region having the amino acid sequence of SEQ ID NO:223.

A nucleic acid that encodes a multispecific antigen-binding molecule that comprises:

• (1) a domain comprising an antibody variable region having glypican 3-binding activity,
• (2) a domain comprising an antibody variable region having T-cell receptor complex-binding activity, and
• (3) a domain comprising an Fc region with reduced binding activity towards an Fcγ receptor,
• wherein the antibody variable region of (1) and the antibody variable region of (2) have a common light chain amino acid sequence;
• and wherein the multispecific antigen-binding molecule has a cytotoxic activity equivalent to or greater than the cytotoxic activity of a bispecific antibody having a glypican 3-binding domain comprising SEQ ID NOs:
• 47 and 48, and a T-cell receptor complex-binding domain comprising SEQ ID NOs:
• 49 and 50.