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 method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a humanized or chimeric bispecific antibody comprising a first binding region and a second binding region, wherein the first binding region binds to human CD3and comprises a heavy chain variable (VH) region comprising VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable (VL) region comprising the amino acid sequence of SEQ ID NO: 10, but wherein:
(i) the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is not T;
(ii) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and wherein one or more of the amino acid positions corresponding to the positions T41, K55, and L97 in SEQ ID NO: 10 are not T, K, and L, respectively;
(iii) the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are not R and A, respectively;
(iv) the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are not F, R, A, R, D, A, D, S, I, and F, respectively;
(v) the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K; or
(vi) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K,
wherein the numbering of positions is according to the EU Index, and wherein the second binding region binds to a cancer-specific target, a target that is overexpressed in cancer, or a target associated with cancer.
2. The method of claim 1 , wherein in subpart (i), the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is selected from the group consisting of: H, I, K, L, Q, R, and V.
3. The method of claim 1 , wherein in subpart (ii), the amino acids in the positions corresponding to positions F10, T41, K55, and L97 in SEQ ID NO: 10 are not F, T, K, and L, respectively.
4. The method of claim 3 , wherein the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is selected from the group consisting of: H, I, K, L, Q, R or V, and wherein the amino acids in the positions corresponding to positions F10, K55, and L97 in SEQ ID NO: 10 are L, N, and H, respectively.
5. The method of claim 1 , wherein in subpart (iii), the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are A and P, respectively.
6. The method of claim 1 , wherein in subpart (iv), the amino acid in the position corresponding to position R23 in SEQ ID NO: 10 is selected from the group consisting of A, G, H, K, Q, S, and T, the amino acid in the position corresponding to A35 in SEQ ID NO: 10 is selected from the group consisting of I, L, M, P, V, G, F, and W, and the amino acids in the positions corresponding to positions F10, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are L, T, G, P, E, A, E, and Y, respectively.
7. The method of claim 6 , wherein the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are L, A, P, T, G, P, E, A, E, and Y, respectively.
8. The method of claim 1 , wherein in subpart (v), the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is N.
9. The method of claim 1 , wherein in subpart (vi), the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is L, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is N.
10. The method of claim 1 , wherein the VH region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6-9.
11. A method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a humanized or chimeric bispecific antibody comprising a first binding region and a second binding region, wherein the first binding region binds to human CD3 and comprises VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and VL CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NO: 4, GTN, and SEQ ID NO: 5 or 60, respectively,
wherein (i) the amino acids in the positions corresponding to positions F10, T41, K55, and L97 in the VL region of SEQ ID NO: 10 are L, K, N, and H, respectively, or (ii) the amino acid in the position corresponding to position T41 in the VL region of SEQ ID NO: 10 is K, and
wherein said antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the amino acids in the positions corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain of SEQ ID NO: 15 are F, E, and A, respectively, in both the first heavy chain and second heavy chain, and wherein the amino acid in the position corresponding to F405 in the human IgG1 heavy chain of SEQ ID NO: 15 is L in the first heavy chain, and the amino acid in the position corresponding to K409 in the human IgG1 heavy chain of SEQ ID NO: 15 is R in the second heavy chain,
wherein the numbering of positions is according to the EU Index, and wherein the second binding region binds to a cancer-specific target, a target that is overexpressed in cancer, or a target associated with cancer.
12. The method of claim 11 , wherein the VH region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6-9.
13. The method of claim 11 , wherein the cancer is breast cancer, prostate cancer, non-small cell lung cancer, bladder cancer, ovarian cancer, gastric cancer, colorectal cancer, esophageal cancer, squamous cell carcinoma of the head and neck, cervical cancer, pancreatic cancer, testis cancer, malignant melanoma, or a soft-tissue cancer.
14. The method of claim 11 , wherein the cancer is an indolent or aggressive form of B-cell lymphoma, chronic lymphatic leukemia, or acute lymphatic leukemia.
15. The method of claim 11 , wherein the second binding region binds to human epidermal growth factor receptor 2 (HER2), CD19, epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor (EGFR), CD66e, CD33, ephrin type-A receptor 2 (EphA2), or melanoma-associated chondroitin sulfate proteoglycan (MCSP).
16. The method of claim 1 , wherein the cancer is breast cancer, prostate cancer, non-small cell lung cancer, bladder cancer, ovarian cancer, gastric cancer, colorectal cancer, esophageal cancer, squamous cell carcinoma of the head and neck, cervical cancer, pancreatic cancer, testis cancer, malignant melanoma, or a soft-tissue cancer.
17. The method of claim 1 , wherein the cancer is an indolent or aggressive form of B-cell lymphoma, chronic lymphatic leukemia, or acute lymphatic leukemia.
18. The method of claim 1 , wherein the second binding region binds to human epidermal growth factor receptor 2 (HER2), CD19, epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor (EGFR), CD66e, CD33, ephrin type-A receptor 2 (EphA2), or melanoma-associated chondroitin sulfate proteoglycan (MCSP).