Anti-EFNA4 antibody-drug conjugates
The present invention provides for anti-EFNA4 antibody-drug conjugates and methods for preparing and using the same.
1. An antibody-drug conjugate of the formula: Ab-(L-D), wherein:
(a) Ab is an antibody, or antigen-binding fragment thereof, that binds to EFNA4 and comprises a heavy chain variable region comprising three CDRs set forth as SEQ ID NOs: 15, 19, and 23 and a light chain variable region comprising three CDRs set forth as SEQ ID NOs: 29, 33, and 35; and
(b) L-D is a linker-drug moiety, wherein L is a linker and D is a drug, and wherein D is a calicheamicin or calicheamicin derivative.
2. The antibody-drug conjugate of claim 1 , wherein the Ab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 13 and a light chain variable having an amino acid sequence that is at least 90% identical to SEQ ID NO: 27.
3. The antibody-drug conjugate of claim 2 , wherein the Ab comprises a heavy chain variable region set forth as SEQ ID NO: 13 and a light chain variable region set forth as SEQ ID NO: 27.
4. The antibody-drug conjugate of claim 3 , wherein the Ab comprises a IgG1 heavy chain constant region.
5. The antibody-drug conjugate of claim 4 , wherein the Ab comprises a heavy chain set forth as SEQ ID NO: 25.
6. The antibody-drug conjugate of claim 3 , wherein the Ab comprises a kappa light chain constant region.
7. The antibody-drug conjugate of claim 6 wherein the Ab comprises a light chain set forth as SEQ ID NO: 37.
8. The antibody-drug conjugate of claim 3 , wherein the Ab comprises a heavy chain set forth as SEQ ID NO: 25 and a light chain set forth as SEQ ID NO: 37.
9. The antibody-drug conjugate of claim 1 , wherein the D is an N-acetyl derivative of calicheamicin.
10. The antibody-drug conjugate of claim 9 , wherein the D is N-acetyl-y-calicheamicin.
11. The antibody-drug conjugate of claim 10 , wherein the D is N-acetyl-γ-calicheamicin dimethyl hydrazide (CM).
12. The antibody-drug conjugate of claim 1 , wherein the linker L comprises 4-(4′acetylphenoxy)butanoic acid (AcBut).
13. The antibody-drug conjugate of claim 1 , having a DAR from 1 to 12.
14. The antibody-drug conjugate of claim 1 , wherein:
(a) the Ab comprises a heavy chain set forth as SEQ ID NO: 25 and a light chain set forth as SEQ ID NO: 37; and
(b) the L is 4-(4′acetylphenoxy)butanoic acid (AcBut), and the D is N-acetyl-γ-calicheamicin dimethyl hydrazide (CM).
15. A composition comprising a plurality of an antibody-drug conjugate of claim 1 , and optionally a pharmaceutical carrier, wherein the composition has an average DAR within a range of 1 to 12.
16. The composition of claim 15 , wherein the composition has an average DAR within the range of 3 to 5.
17. The composition of claim 16 , wherein the composition has an average DAR within the range of 3 to 4.
18. The composition of claim 16 , wherein the composition has an average DAR within the range of 4 to 5.
19. The composition of claim 15 , wherein the composition has an average DAR of about 4.
20. A composition comprising a plurality of an antibody-drug conjugate of claim 1 , and optionally a pharmaceutical carrier, wherein the composition has at least 50% antibody-drug conjugates having a DAR from 3 to 5.
21. The composition of claim 20 , wherein the composition has at least 60% antibody-drug conjugates having a DAR from 3 to 5.
22. The composition of claim 21 , wherein the composition has at least 70% antibody-drug conjugates having a DAR from 3 to 5.
23. The composition of claim 22 , wherein the composition has at least 75% antibody-drug conjugates having a DAR from 3 to 5.
24. The composition of claim 22 , wherein the composition has about 70% to 80% antibody-drug conjugates having a DAR from 3 to 5.
25. A process for producing an antibody-drug conjugate of any claim 1 comprising:
(a) linking the L to the D to provide the L-D moiety;
(b) conjugating the linker-drug moiety to the antibody; and
(c) purifying the antibody-drug conjugate.
26. A pharmaceutical composition comprising the antibody-drug conjugate of claim 1 and a pharmaceutically acceptable carrier.
27. A process for producing the composition of claim 15 comprising:
(a) preparing the antibody-drug conjugate; and
(b) purifying the composition having an average DAR within a range of 1 to 12.