CLEC9A-based chimeric protein preparation

Abstract

This application provides, in part, pharmaceutical formulations of Clec9A-based chimeric protein complexes and their uses.

Claims

1. A pharmaceutical composition, (a) A C-type lectin domain family 9 member A (Clec9A)-based chimeric protein complex comprising (i) a targeting moiety that specifically binds to Clec9A, (ii) modified human IFNα2, and (iii) a modified Fc domain; (b) with at least one pharmaceutically acceptable buffer containing histidine buffer; (c) comprising at least one pharmaceutically acceptable excipient containing a surfactant, The pharmaceutical composition having a pH in the range of at least about 4.0 to at least about 6.

0.

2. The pharmaceutical composition according to claim 1, wherein the pH is in the range of at least about 4.5 to at least about 5.5, and optionally the pH is about 5.

5.

3. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is formulated for parenteral administration.

4. The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition is formulated for intravenous administration, and optionally the intravenous administration comprises intravenous infusion or bolus injection.

5. The pharmaceutical composition according to any one of the prior claims, wherein the buffering agent is histidine in a concentration of at least about 10 mM to about 25 mM or less, or contains the same.

6. The pharmaceutical composition according to claim 5, wherein the buffering agent contains histidine at a concentration of about 25 mM.

7. The pharmaceutical composition according to any one of the prior claims, wherein the buffering agent comprises L-histidine or L-histidine hydrochloride monohydrate.

8. The pharmaceutical composition according to any one of the prior claims, wherein the surfactant is a polysorbate surfactant optionally selected from polysorbate 20 (PS20) and poloxamer 188 (Pol188), or comprises the same.

9. The pharmaceutical composition according to claim 8, wherein the PS20 is present at a concentration of at least about 0.01% (w / v) to at least about 0.1% (w / v), and optionally present at a concentration of about 0.05% (w / v).

10. The pharmaceutical composition according to claim 8, wherein the Pol 188 is present at a concentration of at least about 0.005% (w / v) to at least about 0.1% (w / v), and optionally the Pol 188 is present at a concentration of about 0.01% (w / v) to at least about 0.05% (w / v).

11. The pharmaceutical composition according to any one of the prior claims, wherein the buffer further comprises an acetate buffer.

12. The pharmaceutical composition according to claim 11, wherein the acetate buffer contains acetic acid.

13. A pharmaceutical composition according to any one of the prior claims, further comprising one or more additional excipients selected from amino acids, isotonic agents, and antioxidants.

14. The pharmaceutical composition according to claim 13, wherein the amino acid excipient is arginine or comprises arginine.

15. The pharmaceutical composition according to claim 14, wherein the arginine is present at a concentration of at least about 10 mM to about 50 mM, and optionally, the arginine is present at a concentration of about 50 mM.

16. The pharmaceutical composition according to claim 13, wherein the isotonic agent is selected from sodium chloride, sucrose, mannitol, sorbitol, and trehalose.

17. The pharmaceutical composition according to claim 16, wherein the isotonic agent is sodium chloride, and optionally the sodium chloride is present at a concentration of at least about 10 mM to about 50 mM, and optionally the sodium chloride is present at a concentration of about 50 mM.

18. The pharmaceutical composition according to claim 13, wherein the antioxidant excipient is selected from methionine and ethylenediaminetetraacetic acid (EDTA).

19. The pharmaceutical composition according to claim 18, wherein the antioxidant is methionine, and optionally the methionine is present at a concentration of at least about 1 mM to about 5 mM, and optionally the methionine is present at a concentration of about 3 mM.

20. The pharmaceutical composition according to any one of the prior claims, wherein the Clec9A-based chimeric protein complex is present at a concentration of at least about 1 mg / mL.

21. The pharmaceutical composition according to any one of the prior claims, wherein the Clec9A-based chimeric protein complex is present at a concentration of at least about 1 mg / mL to at least about 10 mg / mL.

22. The pharmaceutical composition according to any one of the prior claims, wherein the Clec9A-based chimeric protein complex is substantially in the form of a heterodimer.

23. The pharmaceutical composition according to any one of the prior claims, wherein the modified human IFNα2 has an amino acid sequence that is approximately 95% identical to SEQ ID NO: 9 or 10.

24. The pharmaceutical composition according to claim 23, wherein the modified human IFNα2 has an amino acid sequence that is approximately 98% identical to SEQ ID NO: 9 or 10.

25. The pharmaceutical composition according to claim 24, wherein the modified human IFNα2 has an amino acid sequence that is approximately 99% identical to SEQ ID NO: 9 or 10.

26. The pharmaceutical composition according to any one of the prior claims, wherein the modified human IFNα2 has 1 to 3 mutations in the amino acid sequence of SEQ ID NO: 9 or 10.

27. The pharmaceutical composition according to claim 26, wherein the modified human IFNα2 contains R149A with respect to SEQ ID NO: 9 or 10, or contains one of the mutations R33A, R144A, R144I, R144L, R144S, R144T, R144Y, A145D, A145G, A145H, A145K, A145Y, M148A, and L153A, optionally containing A145G with respect to SEQ ID NO: 9 or 10.

28. The pharmaceutical composition according to any one of the prior claims, wherein the chimeric protein complex comprises a recombinant heavy chain antibody (VHH).

29. The pharmaceutical composition according to claim 28, wherein VHH has an amino acid sequence that is at least about 95% identical to one of SEQ ID NOs: 11 or 12.

30. The pharmaceutical composition according to claim 29, wherein VHH has an amino acid sequence that is at least about 98% identical to one of SEQ ID NOs: 11 or 12.

31. The pharmaceutical composition according to claim 30, wherein VHH has an amino acid sequence that is at least about 99% identical to one of SEQ ID NOs: 11 or 12.

32. The cell according to claim 31, wherein the VHH has one of the amino acid sequences of SEQ ID NOs: 11 and 12.

33. The pharmaceutical composition according to any one of the prior claims, wherein the modified Fc domain comprises one or more of the following mutations for any one of Sequence IDs 13 to 16: P329G, K322Q, K322A, P331G, or P331S.

34. The pharmaceutical composition according to claim 33, wherein the modified Fc domain contains one or more of the following mutations relative to human IgG1 FC: P329G, K322Q, K322A, P331G, or P331S.

35. The pharmaceutical composition according to claim 33 or 34, wherein the modified Fc domain has an amino acid sequence that is approximately 90% identical to sequence numbers 13 to 16.

36. The pharmaceutical composition according to claim 33 or 34, wherein the modified Fc domain has an amino acid sequence that is approximately 93% identical to sequence numbers 13 to 16.

37. The pharmaceutical composition according to claim 33 or 34, wherein the modified Fc domain has an amino acid sequence that is approximately 95% identical to sequence numbers 13 to 16.

38. The pharmaceutical composition according to any one of the prior claims, wherein the Clec9A-based chimeric protein complex comprises a polypeptide having an amino acid sequence that is at least about 95%, at least about 98%, or at least about 99% identical to one of SEQ ID NOs: 1-4 and 43.

39. The pharmaceutical composition according to any one of claims 1 to 37, comprising a Clec9A-based chimeric protein complex, an amino acid sequence selected from SEQ ID NOs: 1 to 4 and 43, and a polypeptide having fewer than 10 mutations relative to the amino acid sequence.

40. The pharmaceutical composition according to claim 39, comprising a Clec9A-based chimeric protein complex, an amino acid sequence selected from SEQ ID NOs: 1-4 and 43, and a polypeptide having fewer than five mutations relative to the amino acid sequence.

41. The pharmaceutical composition according to any one of claims 1 to 38, wherein the Clec9A-based chimeric protein complex comprises a polypeptide having an amino acid sequence selected from SEQ ID NOs: 1 to 4 and 43.

42. The pharmaceutical composition according to any one of the prior claims, wherein the Clec9A-based chimeric protein complex comprises a polypeptide having an amino acid sequence that is at least about 95% identical to any one of SEQ ID NOs. 5-8, 29-36, or 41-42.

43. The pharmaceutical composition according to claim 42, wherein the Clec9A-based chimeric protein complex comprises a polypeptide having an amino acid sequence that is at least about 98% identical to any one of SEQ ID NOs. 5-8, 29-36, or 41-42.

44. The pharmaceutical composition according to claim 43, wherein the Clec9A-based chimeric protein complex comprises a polypeptide having an amino acid sequence that is at least about 99% identical to any one of SEQ ID NOs. 5-8, 29-36, or 41-43.

45. The aforementioned Clec9A-based chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 95% identical to one of Sequence ID No. 1 or 3, and (ii) A pharmaceutical composition according to any one of claims 42 to 44, comprising a polypeptide having an amino acid sequence having at least about 95% identity with either SEQ ID NO: 5 or 6.

46. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 1 or 3, and (ii) The pharmaceutical composition according to claim 45, comprising a polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 5 or 6.

47. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 1 or 3, and (ii) The pharmaceutical composition according to claim 46, comprising a polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 5 or 6.

48. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 95% identical to either SEQ ID NO: 2 or 4, and (ii) A pharmaceutical composition according to any one of claims 42 to 44, comprising a polypeptide having an amino acid sequence having at least about 95% identity with either SEQ ID NO: 7 or 8.

49. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 2 or 4, and (ii) The pharmaceutical composition according to claim 48, comprising a polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 7 or 8.

50. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 2 or 4, and (ii) The pharmaceutical composition according to claim 48, comprising a polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 7 or 8.

51. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 95% identical to Sequence ID No. 2, and (ii) A pharmaceutical composition according to any one of claims 42 to 44, comprising a polypeptide having an amino acid sequence that is at least about 95% identical to either SEQ ID NO: 31 or 32.

52. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 98% identical to Sequence ID No. 2, and (ii) The pharmaceutical composition according to claim 51, comprising a polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 31 or 32.

53. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 99% identical to Sequence ID No. 2, and (ii) The pharmaceutical composition according to claim 52, comprising a polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 31 or 32.

54. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 95% identical to Sequence ID No. 43, and (ii) The pharmaceutical composition according to any one of claims 42 to 44, comprising a polypeptide having an amino acid sequence having at least about 95% identity with either SEQ ID NO: 41 or 42.

55. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 98% identical to Sequence ID No. 43, and (ii) The pharmaceutical composition according to claim 54, comprising a polypeptide having an amino acid sequence that is at least about 98% identical to either SEQ ID NO: 41 or 42.

56. The aforementioned chimeric protein complex (i) A polypeptide having an amino acid sequence that is at least about 99% identical to Sequence ID No. 43, and (ii) The pharmaceutical composition according to claim 55, comprising a polypeptide having an amino acid sequence that is at least about 99% identical to either SEQ ID NO: 41 or 42.

57. The pharmaceutical composition according to any one of the prior claims, wherein the pharmaceutical composition is freeze-dried.

58. The pharmaceutical composition according to any one of the prior claims, wherein the pharmaceutical composition forms a freeze-dried powder.

59. The pharmaceutical composition according to any one of the prior claims, wherein the pharmaceutical composition is contained in an injection device, and optionally the injection device is a syringe or an injection pen.

60. A kit comprising the freeze-dried powder, vial, and syringe according to claim 58.

61. A method for treating or preventing cancer, comprising administering an effective amount of the composition according to any one of claims 1 to 59 to a patient in need thereof.

62. The method according to claim 61, wherein the cancer is a solid tumor.

63. The method according to claim 61, wherein the cancer is a blood cancer.

64. The aforementioned cancers include basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system cancer, breast cancer, peritoneal cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer (including gastrointestinal cancer), glioblastoma, liver cancer, hepatocellular carcinoma, neoplasm in situ, kidney cancer or renal cancer, laryngeal cancer, leukemia, liver cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma), (and squamous cell carcinoma of the lung), melanoma, myeloma, neuroblastoma, oral cancer (lips, tongue, mouth, pharynx), ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, cancers of the respiratory system, salivary gland cancer, sarcoma (e.g., Kaposi's sarcoma), skin cancer, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, uterine or endometrial cancer, cancers of the urinary system, vulvar cancer, Hodgkin lymphoma and non-Hodgkin lymphoma, B-cell lymphoma The method according to any one of claims 61 to 63, comprising lymphomas including (low-grade / follicular non-Hodgkin lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-incisional cell NHL, giant lesion NHL, mantle cell lymphoma, AIDS-related lymphoma), and Waldenström macroglobulinemia, chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, and other carcinomas and sarcomas, post-transplant lymphoproliferative disorders (PTLD), and nevus syndromes, abnormal angiogenesis associated with edema (e.g., those associated with brain tumors), and Meigs syndrome.

65. A pharmaceutical composition, (a) a C-type lectin domain family 9 member A (Clec9A)-based chimeric protein complex comprising (i) a targeting moiety that specifically binds to Clec9A, (ii) a modified human IFNα2, and (iii) a modified Fc domain, wherein the Clec9A-based chimeric protein complex is present at a concentration of at least about 1 mg / mL; (b) A buffer containing a histidine buffer, wherein histidine is present in a concentration in the range of about 10 mM to about 25 mM; (c) an excipient comprising a surfactant selected from PS20 in a concentration ranging from approximately 0.01% to approximately 0.1%, and Pol188 in a concentration ranging from approximately 0.01% to approximately 0.1%; (d) an amino acid excipient containing arginine, wherein arginine is present in a concentration of at least about 10 mM to about 50 mM or less; (e) an antioxidant excipient containing methionine, wherein methionine is present at a concentration of at least about 1 mM to at least about 5 mM; (f) an isotonic agent containing sodium chloride, wherein sodium chloride is present at a concentration of at least about 10 mM to about 50 mM or less, comprising the isotonic agent, The pharmaceutical composition having a pH in the range of at least 4.5 to at least about 5.

5.

66. A pharmaceutical composition, (a) a C-type lectin domain family 9 member A (Clec9A)-based chimeric protein complex comprising (i) a targeting moiety that specifically binds to Clec9A, (ii) a modified human IFNα2, and (iii) a modified Fc domain, wherein the Clec9A-based chimeric protein complex is present at a concentration of at least about 1 mg / mL to at least about 10 mg / mL; (b) A buffer containing a histidine buffer, wherein histidine is present at a concentration of about 25 mM; (c) an excipient containing a surfactant, Pol 188, at a concentration of approximately 0.05%; (d) an amino acid excipient containing arginine at a concentration of approximately 50 mM; (e) With an antioxidant excipient containing methionine at a concentration of approximately 3 mM; (f) an isotonic agent containing sodium chloride at a concentration of approximately 40 mM, The pharmaceutical composition having a pH of approximately 5.5.

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