Methods and compositions related to long half-life coagulation complexes

• US 10,596,232 B2
• Filed: 08/10/2016
• Issued: 03/24/2020
• Est. Priority Date: 08/12/2015
• Status: Active Grant

• Current Assignee: Cell Machines, Inc.
• Original Assignee: Cell Machines, Inc.
• Inventors: Kelly, James
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US15/751,239
• Publication Number:
  US 20180228872A1
• Time in Patent Office: 1,322 Days
• CPC Class Codes: A61K 38/36 Blood coagulation or fibrinolysis factors
• CPC Class Codes: A61K 38/37 Factors VIII
• CPC Class Codes: A61K 38/48 acting on peptide bonds (3.4)
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/643 Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C12N 9/6437 Coagulation factor VIIa (3.4.21.21)
• CPC Class Codes: C12Y 304/21021 Coagulation factor VIIa (3.4.21.21)

Abstract

Disclosed herein are methods and compositions related to coagulation factor complexes comprising a coagulation factor; a fusion protein; and a modifying molecule, wherein the modifying molecule is coupled to the coagulation factor in such a way as to allow binding by the fusion protein, thereby creating a modified coagulation factor; wherein the modified coagulation factor and the fusion protein interact in at least two independent sites.

Claims

1. A coagulation factor complex comprising:

a. a coagulation factor;

b. a fusion protein comprising a first protein fused to albumin or an albumin fragment; and

c. a modifying molecule, wherein the modifying molecule is coupled to the coagulation factor in such a way as to allow binding by the fusion protein, thereby creating a modified coagulation factor, wherein the modifying molecule is attached to the coagulation factor through at least one bond that is not a peptide bond;

wherein the modified coagulation factor and the fusion protein interact in at least two independent sites.

2. The coagulation factor complex of claim 1, wherein at least one binding site of the modified coagulation factor is a natural binding site.

3. The coagulation factor complex of claim 1, wherein at least one binding site of the fusion protein is provided by the modifying molecule.

4. The coagulation factor complex of claim 1, wherein the coagulation factor is Factor VIII.

5. The coagulation factor complex of claim 1, wherein the first protein of the fusion protein is a fragment of von Willebrand'"'"'s factor.

6. The coagulation factor complex of claim 5, wherein the fragment of von Willebrand'"'"'s factor is a D′

D3 fragment.

7. The coagulation factor complex of claim 1, wherein the modified coagulation factor and the fusion protein interact covalently.

8. The coagulation factor complex of claim 7, wherein the covalent bond is not a peptide bond.

9. The coagulation factor complex of claim 7, wherein the covalent bond is not a peptide bond produced by translation of a nucleic acid.

10. The coagulation factor complex of claim 1, wherein the modifying molecule comprises a fatty acid.

11. The coagulation factor complex of claim 1, wherein the modifying molecule is attached to the coagulation factor through a polyethylene glycol chain.

12. The coagulation factor complex of claim 11, wherein the polyethylene glycol chain is linked to a maleimide on the coagulation factor.

13. The coagulation factor complex of claim 12, wherein the polyethylene glycol chain is linked to maleimide with a structure comprising maleimide-PEG_N-X, wherein this X is methyl tetrazine.

14. The coagulation factor complex of claim 1, wherein the first protein of the fusion protein is joined together with albumin via a linker.

15. The coagulation factor complex of claim 1, wherein the coagulation factor comprises modified amino acids.

16. The coagulation factor complex of claim 1, wherein the fusion protein comprises modified amino acids.

17. The coagulation factor complex of claim 1, wherein half-life of the coagulation factor complex is at least 20% greater compared to the coagulation factor alone.

18. A kit comprising the coagulation factor complex of claim 1.

19. A method of treating a subject with a disease requiring coagulation factor infusion, the method comprising administering to the subject the coagulation factor complex of claim 1.

20. The method of claim 19, wherein the administration of the coagulation factor complex to the subject results in a blood level half-life of the coagulation factor complex which is greater than the blood level half-life obtained upon administration of the coagulation factor alone.

21. A coagulation factor complex comprising:

a. a coagulation factor;

b. a fusion protein comprising a first protein fused to albumin or an albumin fragment; and

c. a modifying molecule, wherein the modifying molecule is coupled to the coagulation factor in such a way as to allow binding by the fusion protein, thereby creating a modified coagulation factor, wherein the modifying molecule is attached to the coagulation factor through a polyethylene glycol chain;

wherein the modified coagulation factor and the fusion protein interact in at least two independent sites.

22. The coagulation factor complex of claim 21, wherein the coagulation factor is Factor VIII.

23. The coagulation factor complex of claim 21, wherein the first protein of the fusion protein is a fragment of von Willebrand'"'"'s factor.