Humanized or chimeric CD3 antibodies
The present invention relates to humanized or chimeric antibodies binding CD3. It furthermore relates to bispecific antibodies, compositions, pharmaceutical compositions, use of said antibodies in the treatment of a disease, and method of treatment.
1. A humanized antibody which binds to human CD3, wherein said antibody comprises a heavy chain variable (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 6, and a light chain variable (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 10.
2. The antibody according to claim 1 , wherein the antibody is a full-length antibody.
3. The antibody according to claim 1 , wherein the antibody comprises an Fc region comprising a first and a second immunoglobulin heavy chain.
4. The antibody according to claim 1 , wherein the first and the second heavy chains are of an isotype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.
5. The antibody according to claim 1 , wherein the antibody comprises an Fc region which has been modified so that:
(a) binding of C1q to the antibody is reduced compared to a wild-type antibody by at least 70%, wherein C1q binding is determined by ELISA;
(b) the antibody mediates reduced Fc-mediated T-cell proliferation compared to a wild-type antibody by at least 50%, wherein said T-cell proliferation is measured in a peripheral blood mononuclear cell (PBMC)-based functional assay; or
(c) the antibody reduces Fc-mediated CD69 expression by at least 50%, when compared to a wild-type antibody wherein said Fc-mediated CD69 expression is determined in a PBMC-based functional assay.
6. The antibody according to claim 1 , wherein the antibody comprises a first and a second immunoglobulin heavy chain, wherein in at least one of the first and second immunoglobulin 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, are not L, L, D, N, and P, respectively, and wherein the amino acid positions are numbered according to the EU numbering system.
7. The antibody according to claim 6 , wherein in
(a) at least one of the first and second heavy chains the amino acids in the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain, are F and E; or A and A, respectively;
(b) at least one of the first and second heavy chains the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are F, E, and A; or A, A, and A, respectively; or
(c) at least one of said first and second heavy chains the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are A, A, and A, respectively; and
(d) at least one of said first and second heavy chains the amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain, are F, E, A, Q, and S, respectively.
8. The antibody according to claim 6 , wherein in at least one of the first and second heavy chains the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are F, E, and A, respectively.
9. A bispecific antibody comprising a first binding region of an antibody according to claim 1 , and a second binding region which binds a different target than the first antigen binding region.
10. The bispecific antibody according to claim 9 , wherein each of the first and second heavy chain comprises at least a hinge region, a CH2 and CH3 region, wherein in the first heavy chain, at least one of the amino acids in the positions corresponding to a positions selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and in the second heavy chain, at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and wherein the first and said second heavy chains are not substituted in the same positions.
11. The bispecific antibody according to claim 10 , wherein the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second heavy chain, or vice versa.
12. A composition comprising the antibody of claim 1 and a carrier.
13. A kit for detecting the presence of CD3 antigen, or a cell expressing CD3, in a sample comprising:
i) the antibody of claim 1 ; and
ii) instructions for use of said kit.
14. The antibody according to claim 8 , wherein in said first and second heavy chains, the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are F, E, and A, respectively, and the amino acids in the positions corresponding to N297 and P331 in a human IgG1 heavy chain, are N and P, respectively.
15. The antibody according to claim 14 , which comprises heavy chain constant regions of SEQ ID NO: 16.
16. The antibody according to claim 14 , which comprises heavy chain constant regions of SEQ ID NO: 25.
17. The antibody according to claim 14 , which comprises heavy chain constant regions of SEQ ID NO: 26.
18. The bispecific antibody according to claim 9 , which comprises a first and a second immunoglobulin heavy chain, wherein in at least one of the first and second immunoglobulin 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, are not L, L, D, N, and P, respectively, and wherein the amino acid positions are numbered according to the EU numbering system.
19. The bispecific antibody according to claim 18 , wherein in said first and second heavy chains the amino acids in the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain, are F, E, and A, respectively, and the amino acids in the positions corresponding to N297 and P331 in a human IgG1 heavy chain, are N and P, respectively.