Multispecific antibodies against CD40 and CD137

1. A multispecific antibody 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 sequence as set forth in SEQ ID NO: 1, an HCDR2 sequence comprising the amino sequence as set forth in SEQ ID NO: 2, and an HCDR3 sequence comprising the amino sequence as set forth in SEQ ID NO: 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino sequence as set forth in SEQ ID NO: 4, a LCDR2 sequence comprising the amino sequence YTS, and a LCDR3 sequence comprising the amino 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 sequence as set forth in SEQ ID NO: 64, an HCDR2 sequence comprising the amino sequence as set forth in SEQ ID NO: 65, and an HCDR3 sequence comprising the amino sequence as set forth in SEQ ID NO: 66, and a second VL region comprising a LCDR1 sequence comprising the amino sequence as set forth in SEQ ID NO: 67, a LCDR2 sequence comprising the amino sequence GAS, and a LCDR3 sequence comprising the amino 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 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; and

wherein (i) the amino acid in the position corresponding to F405 in the human IgG1 heavy chain according to EU numbering is L in the first heavy chain, and the amino acid position corresponding to K409 in the human IgG1 heavy chain according to EU numbering is R in the second heavy chain, or (ii) the amino acid in the position corresponding to K409 in the human IgG1 heavy chain according to EU numbering is R in the first heavy chain, and the amino acid position corresponding to F405 in the human IgG1 heavy chain according to EU numbering is L in the second heavy chain.

2. The multispecific antibody according to claim 1 , wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117, and the first VL region comprises the amino acid sequence as set forth in SEQ ID NO:121, and wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123, and the second VL region comprises the amino sequence as set forth in SEQ ID NO: 127.

3. The multispecific antibody according to claim 1 , wherein the amino acid in the position corresponding to K409 in the human IgG1 heavy chain according to EU numbering is R in the first heavy chain, and the amino acid position corresponding to F405 in the human IgG1 heavy chain according to EU numbering is L in the second heavy chain.

4. The multispecific antibody according to claim 1 , wherein the amino acid in the position corresponding to K409 in the human IgG1 heavy chain according to EU numbering is L in the first heavy chain, and the amino acid position corresponding to F405 in the human IgG1 heavy chain according to EU numbering is R in the second heavy chain.

5. The multispecific antibody of claim 1 , wherein the first heavy chain comprises the amino acid sequence as set forth in SEQ ID NO:119 and the first light chain comprises the amino acid sequence as set forth in SEQ ID NO:122, and wherein the second heavy chain comprises the amino acid sequence as set forth in SEQ ID NO:125 and the second light chain comprises the amino acid sequence as set forth in SEQ ID NO:128.

6. The multispecific antibody of claim 1 , wherein the first heavy chain comprises the amino acid sequence as set forth in SEQ ID NO:119 and the first light chain comprises the amino acid sequence as set forth in SEQ ID NO:122.

7. The multispecific antibody of claim 1 , wherein the second heavy chain comprises the amino acid sequence as set forth in SEQ ID NO:125 and the second light chain comprises the amino acid sequence as set forth in SEQ ID NO:128.

8. The multispecific antibody according to claim 1 , wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117.

9. The multispecific antibody according to claim 1 , wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117, and the first VL region comprises the amino acid sequence as set forth in SEQ ID NO:121.

10. The multispecific antibody according to claim 1 , wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123.

11. The multispecific antibody according to claim 1 , wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123, and the second VL region comprises the amino sequence as set forth in SEQ ID NO: 127.