Multispecific antibodies against CD40 and CD137

A multispecific antibody or a fragment thereof comprising a first heavy chain and a first light chain comprising a first antigen-binding region capable of binding to human CD40 and a second heavy chain and a second light chain comprising a second antigen-binding region capable of binding to human CD137,

• wherein the first antigen-binding region comprises a first heavy chain variable (VH) region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 1, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 2, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 4, a LCDR2 sequence comprising the amino acid sequence YTS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 5;
• wherein the second antigen-binding region comprises a second VH region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 64, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 65, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 66, and a second VL region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 67, a LCDR2 sequence comprising the amino acid sequence GAS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 68,
• wherein the positions corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain according to EU numbering are F, E, and A, respectively, in the first and second heavy chains.

A multispecific antibody or a fragment thereof comprising a first heavy chain and a first light chain comprising a first antigen-binding region capable of binding to human CD40, and a second heavy chain and a second light chain comprising a second antigen-binding region capable of binding to human CD137,

• wherein the first antigen-binding region capable of binding to human CD40 comprises a first heavy chain variable (VH) region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 1, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 2, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 4, a LCDR2 sequence comprising the amino acid sequence YTS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 5;
• wherein the second antigen-binding region capable of binding to human CD137 comprises a second VH region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 64, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 65, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 66, and a second VL region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 67, a LCDR2 sequence comprising the amino acid sequence GAS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO:
• 68;
• wherein the first and second heavy chains each comprise an amino acid substitution at one or more positions corresponding to EU index positions T366, L368, K370, D399, F405, Y407, and/or K409 in a human IgG1 heavy chain, and wherein the first and second heavy chains are not substituted in the same positions,
• wherein in at least one of said first and second heavy chains one or more amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively.