RECOMBINANT OR TRANSGENIC FACTOR VII COMPOUND HAVING A MAJORITY OF GLYCAN, BIANTENNARY, BISIALYLATED AND NON-FUCOSYLATED FORMS

• US 20090239788A1
• Filed: 07/31/2007
• Published: 09/24/2009
• Est. Priority Date: 08/01/2006
• Status: Abandoned Application

• Current Assignee: Lfb Biomedicaments (Agence des Participations de l’Etat)
• Original Assignee: Lfb Biomedicaments (Agence des Participations de l’Etat)
• Inventors: Nony, Emmanuel, Bihoreau, Nicolas, Chtourou, Abdessatar Sami
• Application Number:
  US12/374,269
• Publication Number:
  US 20090239788A1
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/107 Rabbit
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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: C12N 9/64 derived from animal tissue
• CPC Class Codes: C12N 9/6437 Coagulation factor VIIa (3.4.21.21)
• CPC Class Codes: C12N 9/647 Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups
• CPC Class Codes: C12Y 304/21021 Coagulation factor VIIa (3.4.21.21)

Abstract

The present invention concerns a recombinant or transgenic factor VII compound, each factor VII molecule of the compound having glycan forms linked to N-glycosylation sites, wherein among all the factor VII molecules in said compound, glycan, biantennary, bisialylated and non-fucosylated forms are in the majority. The invention also concerns such a compound for use as a medication, and a method for preparing said compound, among others.

Claims

1. A composition of recombinant or transgenic Factor VII (FVII), each molecule of Factor VII of the composition containing glycan forms bound to N-glycosylation sites, wherein among all the molecules of Factor VII of said composition the majority are biantennary, bisialylated and non fucosylated glycan forms compared to all glycan forms bound to N-glycosylation sites of Factor VII of the composition.

2. A composition according to claim 1, the rate of biantennary, bisialylated, fucosylated and non fucosylated forms is higher than 50%.

3. A composition according to claim 1, wherein among all the molecules of Factor VII of said composition, the rate of fucose is comprised between 20% and 50%.

4. A composition according to claim 1, wherein at least some of the sialic acids of Factor VII of said composition imply α

2-6-links.

5. A composition according to claim 4, wherein all sialic acids of Factor VII of said composition imply α

2-6-links.

6. A composition according to claim 4, wherein Factor VII of said composition comprises moreover sialic acids of α

2-3-links.

7. A composition according to claim 1, wherein the said FVII is activated.

8. A composition according to claim 1, for the use as medicament.

9. The use of a composition of Factor VII according to claim 1, for preparing a medicament intended for the treatment of patients suffering from haemophilia.

10. The use of a composition of Factor VII according to claim 1, for preparing a medicament intended for the treatment of multiple hemorragic traumas.

11. The use of a composition of Factor VII according to claim 1, for preparing a medicament intended for the treatment of bleedings due to an overdose of anticoagulants.

12. A pharmaceutical composition comprising a FVII as defined according to claim 1, compromising an excipient and/or a pharmaceutically acceptable carrier.

13. A process for preparing a composition of recombinant or transgenic Factor VII, each molecule of Factor VII of the composition comprises glycan forms bound to N-glycosylation sites and wherein among all molecules of Factor VII of said composition the majority are biantennary, bisialylated glycan forms, the process comprising a step of sialylation by contacting a composition of partially sialylated transgenic or recombinant Factor VII with a sialic acid donor substrate and a sialyltransferase, in a suitable reaction medium in order to allow the activity of the sialyltransferase, for a sufficient period of time and under suitable conditions in order to allow a transfer of the sialic acid from the sialic acid donor substrate to FVII and a sufficient increase in bisialylated forms so that the said bisialylated forms become majority.

14. A process according to claim 13, wherein, prior to the step of sialylation, a step of galactosylation is performed, comprising grafting a galactose on galactose-deficient forms representing the agalactosylated and monogalactosylated forms of FVII.

15. A process according to claim 13, wherein the said composition of partially sialylated FVII exhibits majority biantennary, monosialylated glycan forms.

16. A process according to claim 15, wherein among the biantennary, monosialylated glycan forms of said composition of partially sialylated FVII, the majority glycan forms are non fucosylated.

17. A process according to claim 13, wherein the said composition of partially sialylated FVII exhibits at least some of sialic acids implying α

2-6-links.

18. A process according to claim 13, comprising prior to the sialylation step, a step of production of the composition of partially sialylated transgenic FVII by transgenic female rabbits.

19. A process according to claim 13, wherein the FVII of said composition of partially sialylated FVII is activated.

20. A process according to claim 13, wherein said sialyltransferase is the α

2,6-(N)-sialyltransferase, and in that the sialic acid donor group is the cytidine-5′

-monophospho-N-acetyl-neuraminic acid.