Highly purified epcoritamab compositions

Abstract

The present disclosure provides highly purified epcoritamab compositions, which comprise a bispecific antibody that binds CD3 and CD20. In the highly purified compositions, impurities, such as epcoritamab protein variants, process-related impurities and / or product- related impurities, have been minimized or removed. Methods of preparing the highly purified epcoritamab compositions are also provided.

Description

Abstract This disclosure provides a highly purified icoretuzumab composition comprising a bispecific antibody binding to CD3 and CD20. In the highly purified composition, impurities such as icoretuzumab protein variants, process-related impurities, and / or product-related impurities have been minimized or removed. A method for preparing the highly purified icoretuzumab composition is also provided.

Claims

CLAIMS 1. A composition comprising at least 95% epcoritamab and less than 5% of an impurity.

2. The composition of claim 1, wherein epcoritamab comprises: (i) a CD3 binding arm comprising a heavy chain sequence as shown in SEQ ID NO: 1 and a light chain sequence as shown in SEQ ID NO: 2; and (ii) a CD20 binding arm comprising a heavy chain sequence as shown in SEQ ID NO: 3 and a light chain sequence as shown in SEQ ID NO:

4.

3. The composition of claim 1, wherein the composition comprises less than 3% of the impurity.

4. The composition of any one of claims 1-3, wherein the impurity is a product- related impurity.

5. The composition of claim 4, wherein the product-related impurity is selected from the group consisting of a high molecular weight species (HMWS) of epcoritamab, a low molecular weight species (LMWS) of epcoritamab, a charge variant of epcoritamab, an acidic species of epcoritamab, a basic species of epcoritamab, a deamidated species of epcoritamab, a glycosylated species of epcoritamab, a glycated species of epcoritamab, an oxidized species of epcoritamab, an N-terminal modification of epcoritamab, an aggregate of epcoritamab, a fragment of epcoritamab, a residual signal peptide modification of epcoritamab, an Fc fragment of epcoritamab and a Fab fragment of epcoritamab.

6. The composition of claim 4, wherein the product-related impurity is a high molecular weight species (HMWS) of epcoritamab.

7. The composition of claim 6, wherein the HMWS is an epcoritamab dimer comprising a signal sequence as shown in SEQ ID NO:

9.

8. The composition of claim 4, wherein the product-related impurity is a low molecular weight species (LMWS) of epcoritamab.

9. The composition of claim 8, wherein the LWMS is an Fc + Fab fragment of epcoritamab.

10. The composition of claim 4, wherein the product-related impurity is a deamidated acidic species of epcoritamab.

11. The composition of claim 10, wherein the deamidated acidic species of epcoritamab comprises deamidation of one or more residues selected from the group consisting of a N103 residue in the heavy chain of the CD3 binding arm of epcoritamab, a N106 residue in the heavy chain of the CD3 binding arm of epcoritamab, a N103 and a N106 residue in the heavy chain of the CD3 binding arm of epcoritamab, a N333 residue in the heavy chain of the CD3 binding arm of epcoritamab, a N330 residue in the heavy chain of the CD20 binding arm chain of epcoritamab, and combinations thereof.

12. The composition of any one of claims 1-3, wherein the impurity is a process- related impurity.

13. The composition of claim 12, wherein the process-related impurity is selected from the group consisting of a reducing agent, an oxidizing agent, a manufacturing intermediate, a residual IgG1 homodimer intermediate, a residual host cell protein, a host cell nucleic acid, a media component and a chromatographic material.

14. The composition of claim 13, wherein the process-related impurity is a reducing agent.

15. The composition of claim 14, wherein the reducing agent is 2- mercaptoethylamine (2-MEA).

16. The composition of claim 15, wherein 2-MEA is present in the composition at a level at least 10-fold lower than the Permitted Daily Exposure (PDE) for 2-MEA.

17. The composition of claim 13, wherein the process-related impurity is an oxidizing agent.

18. The composition of claim 17, wherein the oxidizing agent is dehydro-L-Ascorbic Acid (dhAA).

19. The composition of claim 18, wherein dhAA is present in the composition at a level at least 100-fold lower than the Permitted Daily Exposure (PDE) for dhAA.

20. The composition of claim 13, wherein the process-related impurity is a manufacturing intermediate of epcoritamab.

21. The composition of claim 20, wherein the manufacturing intermediate is the 3005a antibody or a portion thereof.

22. The composition of claim 20, wherein the manufacturing intermediate is the 3001d antibody or a portion thereof.

23. The composition of claim 4, wherein the product-related impurity is a glycosylated species of epcoritamab.

24. The composition of claim 23, wherein the glycosylated species comprises glycosylation of one or more residues selected from the group consisting of a N106 residue in the heavy chain of the CD3 binding arm of epcoritamab, a N305 residue in theheavy chain of the CD3 binding arm of epcoritamab, a N302 residue in the heavy chain of the CD20 binding arm of epcoritamab, and combinations thereof.

25. The composition of claim 4, wherein the product-related impurity is a glycated species of epcoritamab.

26. The composition of claim 25, wherein the glycated species comprises glycation of one or more residues selected from the group consisting of a K331 residue in the heavy chain of the CD20 binding arm of epcoritamab, a K334 residue in the heavy chain of the CD3 binding arm chain of epcoritamab, and combinations thereof.

27. The composition of claim 4, wherein the product-related impurity is an oxidized species of epcoritamab.

28. The composition of claim 27, wherein the oxidized species comprises oxidation of one or more residues selected from the group consisting of a M363 residue in the heavy chain of the CD20 binding arm of epcoritamab, a M366 residue in the heavy chain of the CD3 binding arm of epcoritamab, and combinations thereof.

29. The composition of claim 13, wherein the process-related impurity is a residual IgG1 homodimer intermediate.

30. The composition of claim 29, wherein the residual IgG1 homodimer intermediate is a residual 3005a species.

31. The composition of claim 29, wherein the residual IgG1 homodimer intermediate is a residual 3001d species.

32. The composition of claim 29, wherein the residual IgG1 homodimer intermediate is a mixture of a residual 3005a species and a residual 3001d species.

33. The composition of any one of claims 29-32, wherein the composition comprises at least 95% epcoritamab and between 0%-2% of the residual IgG1 homodimer intermediate.

34. The composition of claim 13, wherein the process-related impurity is a residual HCP, and wherein the residual HCP is selected from the group consisting of a polyubiquitin, a ubiquitin, a IgG-binding protein A, a peroxiredoxin-1, a thioredoxin reductase 1 (cytoplasmic), a phospholipase B-like, and combinations thereof.

35. The composition of claim 34, wherein the residual HCP is polyubiquitin.

36. The composition of any one of claims 34-35, wherein the composition comprises at least 95% epcoritamab and between 0-100 parts per million (ppm) of the residual HCP.

37. The composition of claim 34, wherein the residual HCP is between 0-50 ppm.

38. The composition of claim 4, wherein the product-related impurity is an N- terminal modification of epcoritamab.

39. The composition of claim 38, wherein the N-terminal modification is selected from the group consisting of a glutamate to pyroglutamate conversion, a residual signal peptide attached to epcoritamab, and combinations thereof.

40. The composition of claim 39, wherein the glutamate to pyroglutamate conversion occurs in at least one of a heavy chain of the CD3 binding arm of epcoritamab, a heavychain of the CD20 binding arm of epcoritamab and a light chain of the CD20 binding arm of epcoritamab.

41. The composition of claim 40, wherein the glutamine to pyroglutamate conversion occurs in the light chain of the CD3 binding arm of epcoritamab.

42. The composition of claim 39, wherein the residual signal peptide comprises an amino acid sequence selected from the group consisting of (a) Ser-Glu-Ala and (b) Leu- Cys-Thr-Gly-Ser-Glu-Ala (SEQ ID NO: 9).

43. A composition comprising at least 95% epcoritamab and less than 5% of an acidic epcoritamab variant that is deamidated at residues N103 and N106 of CD3 heavy chain of epcoritamab.

44. A composition comprising at least 95% epcoritamab and less than 5% of a high molecular weight species (HMWS) of epcoritamab, wherein the HMWS is an epcoritamab dimer comprising a signal sequence having the amino acid sequence shown in SEQ ID NO:

9.

45. A composition comprising at least 95% epcoritamab and less than 5% of a low molecular weight species (LMWS) of epcoritamab, wherein the LMWS is an Fc + Fab fragment of epcoritamab.

46. The composition of any one of claims 1-45, wherein the composition is a pharmaceutical composition.

47. The composition of claim 46, wherein the composition is a stable liquid composition suitable for subcutaneous administration.

48. The composition of claim 47, wherein the composition is a stable liquid aqueous composition that has never been subjected to prior reconstitution.

49. The composition of claim 47, wherein the composition is a lyophilized composition which is subjected to reconstitution before subcutaneous administration 50. The composition of claim 48 or 49, wherein the composition comprises about 4 to 5 mg / mL of epcoritamab.

51. The composition of claim 48 or 49, wherein the composition comprises about 60 mg / mL of epcoritamab.

52. A method of preparing the composition of any one of claims 1-51, the method comprising: (a) combining a first binding arm of epcoritamab and a second binding arm of epcoritamab to form a combined sample; (b) exposing the combined sample to reducing conditions to form a reduced combined sample; (c) treating the reduced combined sample with an oxidizing agent to form an oxidized sample; and (d) subjecting the oxidized sample to cation exchange chromatography such that the composition is prepared.

53. The method of claim 52, wherein the combined sample is pH-adjusted to pH 7.0- 7.7 before exposure to the reducing conditions.

54. The method of claim 52 or 53, wherein ultrafiltration / diafiltration (UF / DF) is performed after each of steps (b), (c) and (d).

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