Cd19 / cd38 multispecific antibodies

Abstract

Described herein are multispecific binding antibodies with reduced isoelectric focusing point.

Description

Abstract This article describes a multispecific binding antibody with a reduced isoelectric focal point.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A multispecific antibody comprising: a) a CD38 binding moiety comprising: i) a first polypeptide comprising an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1, and a heavy chain constant region wherein the heavy chain constant region lacks a C-terminal lysine residue; and ii) a second polypeptide comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 3; and b) a CD 19 binding moiety comprising: i) a third polypeptide comprising an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2, and a heavy chain constant region wherein the heavy chain constant region lacks a C-terminal lysine residue; and ii) a fourth polypeptide comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 3.

2. The multispecific antibody of claim 1, wherein the heavy chain constant region of the first polypeptide, the third polypeptide, or both the first polypeptide and the third polypeptide comprise a human IgGlor a human IgG4 constant region.

3. The multispecific antibody of claim 1, wherein the polypeptide comprising a light chain variable region further comprises a light chain constant region.

4. The multispecific antibody of any one of claims 1 to 3, wherein the second polypeptide, the fourth polypeptide or both the second polypeptide and the fourth polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 6.

5. The multispecific antibody of any one of claims 1 to 4, wherein the CD38 binding moiety comprises one or more amino acid substitutions that inhibit heavy chain homodimerization of the CD38 binding moiety.

6. The multispecific antibody of claim 5, wherein the CD38 binding moiety comprises a T366W substitution according to EU numbering or T366S / L368A / Y407V substitution to the heavy chain of the CD38 binding moiety according to EU numbering.

7. The multispecific antibody of claim 1 or 2, wherein the CD 19 binding moiety comprises one or more amino acid substitutions that inhibit heavy chain homodimerization of the CD 19 binding moiety.

8. The multispecific antibody of claim 7, wherein the CD19 binding moiety comprises a T366W substitution according to EU numbering or T366S / L368A / Y407V substitution to the heavy chain of the CD 19 binding moiety according to EU numbering.

9. The multispecific antibody of any one of claims 1 to 8, wherein the first polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 4.

10. The multispecific antibody of any one of claims 1 to 8, wherein the third polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 5.

11. A common light chain bispecific antibody comprising: an anti-CD38 heavy chain variable region and an anti-CD38 heavy chain constant region, wherein the anti-CD38 heavy chain variable region comprises: a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 7, a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 8, a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 9, and a negatively-charged amino acid at heavy chain variable region position 1 per Kabat numbering, an anti -CD 19 heavy chain variable region and an anti-CD19 heavy chain constant region, wherein the anti-human-CD19 heavy chain variable region comprises: a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 10, a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 11, a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 12, anda negatively-charged amino acid at heavy chain variable region position 1 per Kabat numbering, a common light chain variable region comprising: a light chain complementarity determining region 1 (LCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 13, a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 14 (AAS), and a light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 15, wherein: the anti-CD38 heavy chain constant region and the anti-CD19 heavy chain constant region each lack a C-terminal lysine residue (e.g., K447 per EU numbering); and the common light chain bispecific antibody comprises an experimental isoelectric point (pl) of less than 9.

12. The common light chain bispecific antibody of claim 11, wherein the common light chain bispecific antibody further comprises a Hydrophobicity (HIC) retention time of less than about 10 minutes.

13. The common light chain bispecific antibody of any one of claims 11-12, wherein the terminal lysine residue is K447 per EU numbering.

14. The common light chain bispecific antibody of any one of claims 11-13, wherein the anti- CD19 heavy chain variable region comprises a serine at position 84 and / or a leucine at position 108 according to Kabat numbering.

15. The common light chain bispecific antibody of any one of claims 11-14, wherein the common light chain variable region comprises a histidine at position 32 according to Kabat numbering.

16. The common light chain bispecific antibody of any one of claims 11-15, wherein the negatively-charged amino acid is glutamic acid.

17. The common light chain bispecific antibody of any one of claims 11-16, wherein the pl is between 8.7 and 9.

18. A pharmaceutical composition comprising the multispecific antibody of any one of claims 1 to 10 or the common light chain bispecific antibody of any one of claims 11 to 17 and a pharmaceutically acceptable excipient, diluent, or carrier.

19. A nucleic acid or plurality of nucleic acids encoding the common light chain bispecific antibody of any one of claims 1 to 10 or the common light chain bispecific antibody of any one of claims 11 to 17.

20. A method of treating a cancer in an individual in need thereof comprising administering to the individual the common light chain bispecific antibody of any one of claims 1 to 10 or the common light chain bispecific antibody of any one of claims 11 to 17, thereby treating the cancer in the individual in need thereof.

21. The method of claim 20, wherein the cancer or tumor is a solid-tissue cancer.

22. The method of claim 21, wherein the solid-tissue cancer comprises breast cancer, prostate cancer, pancreatic cancer, lung cancer, kidney cancer, stomach cancer, esophageal cancer, skin cancer, colorectal cancer, or head and neck cancer.

23. The method of claim 22, wherein the breast cancer is triple negative breast cancer, the lung cancer is non-small cell lung cancer, the head and neck cancer is head and neck squamous cell cancer, the kidney cancer is renal cell carcinoma, the brain cancer is glioblastoma multiforme, or the skin cancer is melanoma.

24. The method of claim 20, wherein the cancer or tumor is a blood cancer.

25. The method of claim 24, wherein the blood cancer is diffuse large B cell lymphoma.

26. The method of claim 24, wherein the blood cancer is myeloma.

27. The method of claim 24, wherein the blood cancer is Burkitt’s lymphoma.

28. The method of claim 24, wherein the blood cancer is B cell lymphoma.

29. The method of claim 28, wherein the B cell lymphoma comprises double hit lymphoma, double expressor lymphoma, or triple hit lymphoma.

30. The method of any one of claims 24 to 29, wherein the blood cancer is relapsed or refractory to treatment.

31. The method of any one of claims 20 to 30, wherein the cancer or tumor associated with CD 19 positive, CD38 high immunosuppressive B cells is a cancer or tumor that comprises CD 19 positive, CD38 high B cell infiltrates.

32. The method of claim 31, wherein the CD 19 positive, CD38 high immunosuppressive B cells express a B cell activation marker.

33. The method of claim 32, wherein the B cell activation marker comprises CD30.

34. The method of any one of claims 20 to 33, wherein the cancer or tumor associated with CD 19 positive, CD38 high B cells expresses PD-L1.

35. The method of any one of claims 22 to 34, wherein the cancer or tumor associated with CD 19 positive, CD38 high B cells is associated with CD20 low or CD20 negative B cells.

36. The method of claim 35, wherein the CD38 high B cells express at least about 30,000 CD38 proteins on the cell surface.

37. The method of claim 35, wherein the CD38 high B cells express at least about 35,000 CD38 proteins on the cell surface.

38. The method of claim 35, wherein the CD38 high B cells express at least about 40,000 CD38 proteins on the cell surface.

39. A method of reducing an experimental isoelectric point of an antibody, the method comprising:(a) mutating a glutamine to a negatively charge amino acid at position 1 of a heavy chain variable region per Kabat numbering; and(b) removing a lysine at the C-terminal position of a heavy chain constant region.

40. The method of claim 39, wherein the negatively charged amino acid is glutamic acid.

41. The method of any one of claims 39-40, wherein the antibody is a common light chain bispecific antibody comprising: an anti-CD38 heavy chain variable region, wherein the anti-CD38 heavy chain variable region comprises: a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 7,a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 8, and a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 9, an anti -CD 19 heavy chain variable region, wherein the anti-human-CD19 heavy chain variable region comprises: a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 10, a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 11, and a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 12, a common light chain variable region comprising: a light chain complementarity determining region 1 (LCDR1) comprising an amino acid sequence set forth in SEQ ID NO: 13, a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence set forth in SEQ ID NO: 14 (AAS), and a light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence set forth in SEQ ID NO: 15.

42. A method of making an antibody, the method comprising harvesting the bispecific antibody of any one of claims 1 to 17 from the supernatant of a cell line comprising a nucleic acid encoding the antibody and subjecting the supernatant to one or more purification steps.