MULTIVALENT HETEROMULTIMER SCAFFOLD DESIGN AND CONSTRUCTS

• US 20120244577A1
• Filed: 03/02/2012
• Published: 09/27/2012
• Est. Priority Date: 03/03/2011
• Status: Active Grant

• Current Assignee: Zymeworks BC, Inc. (Zymeworks Inc,)
• Original Assignee: Zymeworks Inc,
• Inventors: Dixit, Surjit Bhimarao, D'Angelo, Igor Edmundo Paolo, Poon, David Kai Yuen
• Granted Patent: US 9,499,605 B2
• Time in Patent Office: Days
• US Class Current: 435/69.6
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4721 Lipocortins
• CPC Class Codes: C07K 14/485 Epidermal growth factor [EGF], i.e. urogastrone
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 14/76 Albumins
• CPC Class Codes: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C07K 14/79 Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C12N 9/6454 Dibasic site splicing serine proteases, e.g. kexin (3.4.21.61); furin (3.4.21.75) and other proprotein convertases
• CPC Class Codes: C12Y 304/21061 Kexin (3.4.21.61), i.e. proprotein convertase subtilisin/kexin type 9

Abstract

Provided herein are multifunctional heteromer proteins. In specific embodiments is a heteromultimer that comprises: at least two monomeric proteins, wherein each monomeric protein comprises at least one cargo polypeptide, attached to a transporter polypeptide, such that said monomeric proteins associate to form the heteromultimer. These therapeutically novel molecules comprise monomers that function as scaffolds for the conjugation or fusion of therapeutic molecular entities resulting in the creation of bispecific or multivalent molecular species.

Claims

1. A heteromultimer comprising:

at least a first monomeric protein that comprises (i) a first transporter polypeptide; and

(ii) at least one cargo polypeptide and

at least a second monomeric protein that comprises (iii) a second transporter polypeptide and (iv) at least one cargo polypeptide;

wherein each transporter polypeptide is derived from a monomeric protein by segmentation of said monomeric protein and wherein said transporter polypeptides self-assemble to form a quasi-native structure of said monomeric protein or analog thereof.

2. The heteromultimer of claim 1, wherein said heteromultimer is a heterodimer.

3. The heteromultimer of claim 1 wherein at least one transporter polypeptide is not derived from an antibody.

4. The heteromultimer of claim 1, wherein each transporter polypeptide is an albumin derivative.

5. The heteromultimer of claim 4, wherein said albumin is human serum albumin having a sequence of SEQ ID NO:

1 or analog or variant thereof.

6. The heteromultimer of claim 1, wherein said first transporter polypeptide has a sequence comprising SEQ ID NO:

2 or analog or variant thereof, and wherein said second transporter polypeptide has a sequence comprising SEQ ID NO;

3 or analog or variant thereof.

7. The heteromultimer of claim 1, wherein said first transporter polypeptide has a sequence comprising SEQ ID NO:

8 or analog or variant thereof, and wherein said second transporter polypeptide has a sequence comprising SEQ ID NO;

10 or analog or variant thereof.

8. The heteromultimer of claim 1, wherein at least one transporter polypeptide is an allo-albumin derivative.

17. The heteromultimer according to claim 1, wherein the transporter polypeptides are derived from the same protein.

18. The heteromultimer according to claim 1, wherein the transporter polypeptides are derived from different proteins.

22. The heteromultimer of claim 1, wherein at least one cargo polypeptide is an antibody, or fragment or variant thereof.

31. The heteromultimer of claim 1 wherein the at least one cargo polypeptide of said first monomeric protein binds a target antigen, and the at least one cargo polypeptide of said second monomeric protein comprises a toxin moiety.

32. The heteromultimer of claim 1, wherein said at least one cargo polypeptide binds a target antigen, and wherein said target antigen is at least one of a-chain (CD25) of IL-2R, Amyloid beta, anti-EpCAM, anti-CD3, CD 16, CD20, CD22, CD23, CD3, CD4, CD52, CD80, CTLA-4, EGFR, EpCAM, F protein of RSV, G250, glycoprotein IIB/IIIa R, HER2, HER2/neu R, HSP90, IgE antibody, IL-12, IL-23, IL-1b, IL-5, IL-6, RANKL, TNF alpha, TNFR, VEGF-A, glucagon receptor, GLP receptor, and LDL receptor.

33. The heteromultimer of claim 1 wherein at least one cargo polypeptide is an enzyme, hormone, therapeutic polypeptide, antigen, chemotoxin, radiotoxin, cytokine or variant or fragment thereof.

34. The heteromultimer of claim 1, wherein said first monomeric protein and said second monomeric protein comprise the same cargo polypeptide.

44. A host cell comprising nucleic acid encoding the heteromultimer of claim 1.

45. The host cell of claim 44, wherein the nucleic acid encoding the first monomeric protein and the nucleic acid encoding the second monomeric protein are present in a single vector.

46. The host cell of claim 44, wherein the nucleic acid encoding the first monomeric protein and the nucleic acid encoding the second monomeric protein are present in separate vectors.

9-16. -16. (canceled)

19-21. -21. (canceled)

23-30. -30. (canceled)

35-43. -43. (canceled)

47. A method of making heteromultimers from a monomeric protein by segmentation of said monomeric protein to obtain polypeptides such that said polypeptides self-assemble to form said heteromultimer.

48. The method of claim 47, wherein the monomeric protein is albumin.

51. The method of claim 47, wherein the segmentation is performed such that the heteromultimer comprises no covalent bond between the polypeptides.

52. The method of claim 47, wherein the segmentation is performed such that the heteromultimer comprises at least one covalent bond between the polypeptides.

53. A heteromultimer designed by the method of claim 47.

54. A therapeutic scaffold comprising a heteromultimer designed by the method of claim 47.

49-50. -50. (canceled)