Multispecific fab fusion proteins and use thereof
The present invention provides multispecific Fab fusion proteins (MSFP) that specifically bind to CD3 and EpCAM. The present invention further provides uses of the MSFPs for the preparation of pharmaceutical compositions, methods of treating cancer, and kits comprising the MSFPs. Also provided are anti-EpCAM antibodies or antigen-binding fragments thereof.
1. An anti-EpCAM antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) comprising: (1) a HVR-H1 comprising the amino acid sequence of SEQ ID NO:13; (2) a HVR-H2 comprising the amino acid sequence of SEQ ID NO:14; and (3) a HVR-H3 comprising the amino acid sequence of SEQ ID NO:15; and a light chain variable region (VL) comprising: (1) a HVR-L1 comprising the amino acid sequence of SEQ ID NO:16; (2) a HVR-L2 comprising the amino acid sequence of SEQ ID NO:17; and (3) a HVR-L3 comprising the amino acid sequence of SEQ ID NO:18.
2. The anti-EpCAM antibody or antigen-binding fragment thereof according to claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO:19; and/or wherein the VL comprises the amino acid sequence of SEQ ID NO:20.
3. The anti-EpCAM antibody of claim 1 , wherein the anti-EpCAM antibody is a multispecific antibody.
4. The antigen-binding fragment of the anti-EpCAM antibody of claim 1 , wherein the antigen-binding fragment is a single-chain FAT (scFv).
5. The antigen-binding fragment of the anti-EpCAM antibody of claim 4 , wherein the scFv comprises the amino acid sequence of SEQ ID NO:21.
6. A multispecific Fab fusion protein comprising the anti-EpCAM antigen-binding fragment of claim 1 .
7. The multispecific Fab fusion protein of claim 6 , comprising a Fab fragment that specifically binds to CD3, a first copy of the anti-EpCAM antigen-binding fragment, and a second copy of the anti-EpCAM antigen binding fragment; wherein the first copy of the anti-EpCAM antigen-binding fragment is fused to the N-terminus of the VH of the Fab fragment; and wherein the second copy of the anti-EpCAM antigen binding fragment is fused to the N-terminus of the VL of the Fab fragment.
8. The multispecific Fab fusion protein of claim 7 , wherein the Fab fragment specifically binds to the N-terminus of CD3 epsilon.
9. The multispecific Fab fusion protein of claim 8 , wherein the Fab fragment specifically binds to an epitope within amino acids 1-27 of CD3 epsilon.
10. The multispecific Fab fusion protein of claim 9 , wherein the VH of the Fab fragment comprises a HVR-H1 comprising the amino acid sequence of SEQ ID NO:1; a HVR-H2 comprising the amino acid sequence of SEQ ID NO:2; and a HVR-H3 comprising the amino acid sequence of SEQ ID NO:3, and/or the VL of the Fab fragment comprises a HVR-L1 comprising the amino acid sequence of SEQ ID NO:4; a HVR-L2 comprising the amino acid sequence of SEQ ID NO:5; and a HVR-L3 comprising the amino acid sequence of SEQ ID NO:6.
11. The multispecific Fab fusion protein of claim 10 , wherein the VH of the Fab fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:7 and 39-43, and/or wherein the VL of the Fab fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:8 and 44-47.
12. The multispecific Fab fusion protein of claim 7 , wherein the Fab fragment comprises a human immunoglobulin heavy chain constant region 1 (CH1) comprising the amino acid sequence of SEQ ID NO:9.
13. The multispecific Fab fusion protein of claim 7 , wherein the Fab fragment comprises a human lambda light chain constant region (CL) comprising the amino acid sequence of SEQ ID NO:10.
14. The multispecific Fab fusion protein of claim 7 , wherein the CH1 and the CL of the Fab fragment are connected by one or more disulfide bonds.
15. The multispecific Fab fusion protein of claim 7 , comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO:22, and a second polypeptide comprising the amino acid sequence of SEQ ID NO:23.
16. A method of treating a cancer in an individual, comprising administering to the individual an effective amount of the multispecific Fab fusion protein of claim 7 .
17. The method of claim 16 , wherein the multispecific Fab fusion protein is administered intravenously.
18. The method of claim 16 , wherein the cancer is selected from the group consisting of small intestine cancer, colorectal cancer, lung cancer, cervical cancer, liver cancer, gastric cancer, pancreatic cancer, skin cancer, renal cancer, bladder cancer, thyroid cancer, prostate cancer, ovarian cancer, endometrial cancer, breast cancer, bile duct cancer, and head and neck cancer.
19. The method of claim 18 , wherein the cancer is colorectal adenocarcinoma.
20. The method of claim 18 , wherein the cancer is lung adenocarcinoma.