Anti-CEACAM5 antibodies and uses thereof
Antibodies are provided which bind human and Macaca fascicularis CEACAM5 proteins, as well as isolated nucleic acids, vectors and host cells comprising a sequence encoding the antibodies. Also provided are immunoconjugates comprising the antibodies conjugated or linked to a growth-inhibitory agent, and to pharmaceutical compositions comprising the antibodies, or immunoconjugates of the invention. The antibodies or immunoconjugates are used for the treatment of cancer or for diagnostic purposes.
1. An isolated nucleic acid comprising a sequence encoding an antibody or antigen binding fragment thereof that binds to human CEACAM5 protein, wherein said antibody or antigen binding fragment thereof comprises a CDR1-H comprising an amino acid sequence of SEQ ID NO:7, a CDR2-H comprising an amino acid sequence of SEQ ID NO:8, a CDR3-H comprising an amino acid sequence of SEQ ID NO:9, a CDR1-L comprising an amino acid sequence of SEQ ID NO:10, a CDR2-L comprising an amino acid sequence of NTR or an amino acid sequence of NTK, and a CDR3-L comprising an amino acid sequence of SEQ ID NO:12.
2. The isolated nucleic acid according to claim 1 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of sequence SEQ ID NO:33 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:34.
3. The isolated nucleic acid according to claim 1 , wherein said antibody or antigen binding fragment thereof is a chimeric or a humanized antibody.
4. The isolated nucleic acid according to claim 3 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:43 and a light chain comprising an amino acid sequence of SEQ ID NO:44.
5. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:17.
6. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:55.
7. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:23.
8. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:29.
9. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:74 and a light chain variable domain comprising an amino acid of SEQ ID NO:75.
10. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:87 and a light chain comprising an amino acid of SEQ ID NO:88.
11. The isolated nucleic acid according to claim 1 , wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:89 and a light chain comprising an amino acid of SEQ ID NO:90.
12. A host cell comprising a nucleic acid sequence encoding an antibody or antigen binding fragment thereof that binds to human CEACAM5 protein, wherein said antibody or antigen binding fragment thereof comprises a CDR1-H comprising an amino acid sequence of SEQ ID NO:7, a CDR2-H comprising an amino acid sequence of SEQ ID NO:8, a CDR3-H comprising an amino acid sequence of SEQ ID NO:9, a CDR1-L comprising an amino acid sequence of SEQ ID NO:10, a CDR2-L comprising an amino acid sequence of NTR or an amino acid sequence of NTK, and a CDR3-L comprising an amino acid sequence of SEQ ID NO:12.
13. The host cell according to claim 12 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of sequence SEQ ID NO:33 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:34.
14. The host cell according to claim 12 , wherein said antibody or antigen binding fragment thereof is a chimeric or a humanized antibody.
15. The host cell according to claim 12 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:43 and a light chain comprising an amino acid sequence of SEQ ID NO:44.
16. The host cell according to claim 12 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:17.
17. The host cell according to claim 12 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:55.
18. The host cell according to claim 12 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:23.
19. The host cell according to claim 12 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:29.
20. The host cell according to claim 12 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:74 and a light chain variable domain comprising an amino acid of SEQ ID NO:75.
21. The host cell according to claim 12 , wherein said antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:87 and a light chain comprising an amino acid of SEQ ID NO:88.
22. The host cell according to claim 12 , wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:89 and a light chain comprising an amino acid of SEQ ID NO:90.
23. A method of producing an antibody or antigen binding fragment that binds to human CEACAM5, the method comprising:
(i) culturing a host cell comprising a nucleic acid sequence encoding an antibody or antigen binding fragment thereof wherein said antibody or antigen binding fragment thereof comprises a CDR1-H comprising an amino acid sequence of SEQ ID NO:7, a CDR2-H comprising an amino acid sequence of SEQ ID NO:8, a CDR3-H comprising an amino acid sequence of SEQ ID NO:9, a CDR1-L comprising an amino acid sequence of SEQ ID NO:10, a CDR2-L comprising an amino acid sequence of NTR or an amino acid sequence of NTK, and a CDR3-L comprising an amino acid sequence of SEQ ID NO:12;
(ii) expressing said antibody or antigen binding fragment thereof; and
(iii) recovering the expressed antibody or antigen binding fragment thereof.
24. The method according to claim 23 , wherein said antibody or antigen binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of sequence SEQ ID NO:33 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:34.
25. The method according to claim 23 , wherein said antibody or antigen binding fragment thereof is a chimeric or a humanized antibody.
26. The method according to claim 23 , wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:43 and a light chain comprising an amino acid sequence of SEQ ID NO:44.
27. The method according to claim 23 , wherein the antibody or fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:17.
28. The method according to claim 23 , wherein the antibody or fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:51 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:55.
29. The method according to claim 23 , wherein the antibody or fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:23.
30. The method according to claim 23 , wherein the antibody or fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:5 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO:29.
31. The method according to claim 23 , wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO:74 and a light chain variable domain comprising an amino acid of SEQ ID NO:75.
32. The method according to claim 23 , wherein said antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:87 and a light chain comprising an amino acid of SEQ ID NO:88.
33. The method according to claim 23 , wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:89 and a light chain comprising an amino acid of SEQ ID NO:90.
34. The method according to claim 23 , further comprising producing an immunoconjugate comprising the antibody or fragment thereof by conjugating or linking at least one growth inhibitory agent to the antibody or fragment thereof using a linker, wherein the cytotoxic agent is selected from the group consisting of: a taxoid, vincas, a maytansinoid, maytansinoid analog, a small drug, a tomaymycin derivative, a pyrrolobenzodiazepine derivative, a cryptophycin derivative, a leptomycin derivative, an auristatin analog, a dolastatin analog, a prodrug, topoisomerase II inhibitors, a DNA alkylating agent, an anti-tubulin agent, a CC-1065 analog, and a CC-1065 analog.
35. The method according to claim 23 further comprising producing an immunoconjugate comprising the antibody or fragment thereof by conjugating or linking to the antibody at least one growth inhibitory agent, wherein the growth inhibitory agent is (N 2 ′-deacetyl-N 2 ′-(3-mercapto-1-oxopropyl)-maytansine) (DM1) or N 2 ′-deacetyl-N- 2 ′(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).
36. The method according to claim 35 , wherein the immunoconjugate comprises a linker selected from the group consisting of N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(Pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), and succinimidyl (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC).
37. The method according to claim 36 , wherein the growth inhibitory agent is DM4 and the linker is SPDB.
38. The method according to claim 36 , wherein the immunoconjugate is characterized by a drug-to-antibody ratio (DAR) ranging from 1 to 10.
39. A method of producing an immunoconjugate comprising:
(i) culturing a host cell which has been transformed by an isolated nucleic acid comprising a sequence encoding an antibody or antigen binding fragment thereof wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:87 and a light chain comprising an amino acid of SEQ ID NO:88;
(ii) expressing said antibody or antigen binding fragment thereof;
(iii) recovering the expressed antibody or antigen binding fragment thereof; and
(iv) conjugating or linking at least one growth inhibitory agent to the antibody or fragment thereof using a linker, wherein the growth inhibitory agent is N 2 ′-deacetyl-N- 2 ′(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) and the linker is 4-(Pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), such that an immunconjugate comprising the antibody is formed.
40. The method according to claim 38 , wherein the immunoconjugate is characterized by a drug-to-antibody ratio (DAR) ranging from 1 to 10.