Claims for Patent: 10,370,430

Summary for Patent: 10,370,430

Title:	Recombinant factor VIII proteins
Abstract:	Provided are recombinant Factor VIII proteins, e.g., human Factor VIII proteins with heterologous moieties inserted into flexible permissive loops located in the Factor VIII A domains, while retaining the procoagulant activity of Factor VIII.
Inventor(s):	Kulman; John (Belmont, MA)
Assignee:	BIOVERATIV THERAPEUTICS INC. (Waltham, MA)
Application Number:	14/379,196
Patent Claims:	1. A recombinant FVIII protein comprising: a first polypeptide comprising Formula I: (A1)-a1-(A2)-a2-[B]; and a second polypeptide comprising Formula II: a3-(A3)-(C1); wherein the first polypeptide and the second polypeptide are fused or exist as a heterodimer; wherein, a) A1 is an A1 domain of FVIII; b) A2 is an A2 domain of FVIII; c) [B] is a B domain of FVIII, a fragment thereof, or is deleted; d) A3 is an A3 domain of FVIII; e) C1 is a C1 domain of FVIII; f) a1, a2, and a3 are acidic spacer regions; wherein the A1 domain comprises a permissive loop-1 (A1-1) region and a permissive loop-2 (A1-2) region; wherein the A2 domain comprises a permissive loop-1 (A2-1) region and a permissive loop-2 (A2-2) region; wherein the A3 domain comprises a permissive loop-1 (A3-1) region and a permissive loop-2 (A3-2) region; wherein a first half-life extending and a second half-life extending moiety are inserted within the FVIII protein immediately downstream of amino acids which correspond to amino acids in mature native human FVIII selected from the group consisting of: a amino acids 18 and 1656 of SEQ ID NO:1; b amino acids 40 and 1656 of SEQ ID NO:1; c amino acids 399 and 1656 of SEQ ID NO:1; d amino acids 403 and 1656 of SEQ ID NO:1; e amino acids 1656 and 1720 of SEQ ID NO:1; f amino acids 1656 and 1900 of SEQ ID NO:1; g amino acids 18 and 399 of SEQ ID NO:1; h amino acids 26 and 399 of SEQ ID NO:1; and i amino acids 40 and 399 of SEQ ID NO:1; wherein the first half-life extending moiety or the second half-life extending moiety is selected from: (a) albumin; (b) albumin-binding polypeptide; (c) Fc; (d) PAS; (e) the C-terminal peptide (CTP) of the .beta. subunit of human chorionic gonadotropin; (f) polyethylene glycol (PEG); (g) hydroxyethyl starch (HES); (h) albumin-binding small molecules; (i) a clearance receptor, or fragment thereof, wherein the clearance receptor blocks binding of the recombinant FVIII protein to FVIII clearance receptors; (j) a low-density lipoprotein receptor-related protein 1 (LRP1) or a FVIII-binding fragment thereof; (k) an XTEN; and (l) any combination of (a)-(j); and wherein the recombinant FVIII protein exhibits procoagulant activity. 2. A composition comprising the recombinant FVIII protein of claim 1 and a pharmaceutically acceptable excipient. 3. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 399 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII. 4. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 18 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 399 of SEQ ID NO:1, corresponding to mature native human FVIII. 5. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 26 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 399 of SEQ ID NO:1, corresponding to mature native human FVIII. 6. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 40 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 399 of SEQ ID NO:1, corresponding to mature native human FVIII. 7. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 18 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII. 8. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 40 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII. 9. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1720 of SEQ ID NO:1, corresponding to mature native human FVIII. 10. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 403 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII. 11. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety is inserted immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII; and the second half-life extending moiety is inserted immediately downstream of amino acid 1900 of SEQ ID NO:1, corresponding to mature native human FVIII. 12. A recombinant FVIII protein comprising: a first polypeptide comprising Formula I: (A1)-a1-(A2)-a2-[B]; and a second polypeptide comprising Formula II: a3-(A3)-(C1); wherein the first polypeptide and the second polypeptide are fused or exist as a heterodimer; wherein, a) A1 is an A1 domain of FVIII; b) A2 is an A2 domain of FVIII; c) [B] is a B domain of FVIII, a fragment thereof, or is deleted; d) A3 is an A3 domain of FVIII; e) C1 is a C1 domain of FVIII; f) a1, a2, and a3 are acidic spacer regions; wherein the A1 domain comprises a permissive loop-1 (A1-1) region and a permissive loop-2 (A1-2) region; wherein the A2 domain comprises a permissive loop-1 (A2-1) region and a permissive loop-2 (A2-2) region; wherein the A3 domain comprises a permissive loop-1 (A3-1) region and a permissive loop-2 (A3-2) region; wherein a first half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 26 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein a second half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 403 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein a third half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is selected from: (a) albumin; (b) albumin-binding polypeptide; (c) Fc; (d) PAS; (e) the C-terminal peptide (CTP) of the .beta. subunit of human chorionic gonadotropin; (f) polyethylene glycol (PEG); (g) hydroxyethyl starch (HES); (h) albumin-binding small molecules; (i) a clearance receptor, or fragment thereof, wherein the clearance receptor blocks binding of the recombinant FVIII protein to FVIII clearance receptors; (j) a low-density lipoprotein receptor-related protein 1 (LRP1) or a FVIII-binding fragment thereof; (k) an XTEN; and (l) any combination of (a)-(j); and wherein the recombinant FVIII protein exhibits procoagulant activity. 13. A recombinant FVIII protein comprising: a first polypeptide comprising Formula I: (A1)-a1-(A2)-a2-[B]; and a second polypeptide comprising Formula II: a3-(A3)-(C1); wherein the first polypeptide and the second polypeptide are fused or exist as a heterodimer; wherein, a) A1 is an A1 domain of FVIII; b) A2 is an A2 domain of FVIII; c) [B] is a B domain of FVIII, a fragment thereof, or is deleted; d) A3 is an A3 domain of FVIII; e) C1 is a C1 domain of FVIII; f) a1, a2, and a3 are acidic spacer regions; wherein the A1 domain comprises a permissive loop-1 (A1-1) region and a permissive loop-2 (A1-2) region; wherein the A2 domain comprises a permissive loop-1 (A2-1) region and a permissive loop-2 (A2-2) region; wherein the A3 domain comprises a permissive loop-1 (A3-1) region and a permissive loop-2 (A3-2) region; wherein a first half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 26 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein a second half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 1656 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein a third half-life extending moiety is inserted within the FVIII protein immediately downstream of amino acid 1720 of SEQ ID NO:1, corresponding to mature native human FVIII; wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is selected from: (a) albumin; (b) albumin-binding polypeptide; (c) Fc; (d) PAS; (e) the C-terminal peptide (CTP) of the .beta. subunit of human chorionic gonadotropin; (f) polyethylene glycol (PEG); (g) hydroxyethyl starch (HES); (h) albumin-binding small molecules; (i) a clearance receptor, or fragment thereof, wherein the clearance receptor blocks binding of the recombinant FVIII protein to FVIII clearance receptors; (j) a low-density lipoprotein receptor-related protein 1 (LRP1) or a FVIII-binding fragment thereof; (k) an XTEN; and (l) any combination of (a)-(j); and wherein the recombinant FVIII protein exhibits procoagulant activity. 14. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety or the second half-life extending moiety is an albumin. 15. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety or the second half-life extending moiety is an Fc. 16. The recombinant FVIII protein of claim 1, wherein the first half-life extending moiety or the second half-life extending moiety is an XTEN. 17. The recombinant FVIII protein of claim 12, wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is an albumin. 18. The recombinant FVIII protein of claim 12, wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is an Fc. 19. The recombinant FVIII protein of claim 12, wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is an XTEN. 20. The recombinant FVIII protein of claim 13, wherein the first half-life extending moiety, the second half-life extending moiety, or the third half-life extending moiety is an XTEN.

Details for Patent 10,370,430

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	January 15, 1974	10,370,430	2033-02-15
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	December 27, 1984	10,370,430	2033-02-15
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	February 15, 1985	10,370,430	2033-02-15