Compositions and methods for the treatment of hemophilia A

• US 20040005670A1
• Filed: 05/22/2003
• Published: 01/08/2004
• Est. Priority Date: 05/22/2002
• Status: Active Grant

• Current Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation)
• Original Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation)
• Inventors: High, Katherine A., Camire, Rodney M.
• Granted Patent: US 7,211,558 B2
• Time in Patent Office: Days
• US Class Current: 435/69.6
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)

Abstract

Improved materials and methods for the treatment of Hemophilia A are provided.

Claims

1. An isolated nucleic acid sequence which encodes a biologically active recombinant human factor VIII (FVIII) variant, said FVIII variant being selected from the group of FVIII variants set forth in Table II.

2. The nucleic acid sequence of claim 1, comprising a first DNA segment coding for the amino acids 1 through 740 of SEQ ID NO:

2 and a second DNA segment coding for the amino acids 1690 through 2332 of SEQ ID NO;

2, said segments being interconnected by at least one linker DNA segment coding for a PACE-furin or PACE furin-like cleavage site.

3. The nucleic acid sequence of claim 2, wherein said at least one PACE-furin or PACE furin-like cleavage site is selected from the group consisting of RRRR (SEQ ID NO:

4) and RKRRKR (SEQ ID NO;

     5).

4. The nucleic acid sequence of claim 1, comprising a first DNA segment coding for the amino acids 1 through 730 of SEQ ID NO:

2 and a second DNA segment coding for the amino acids 1690 through 2332 of SEQ ID NO;

2, said segments being interconnected by at least one linker DNA segment coding for a PACE-furin or PACE furin-like cleavage site.

5. The nucleic acid sequence of claim 4, wherein said at least one PACE-furin or PACE furin-like cleavage site is selected from the group consisting of RRRR (SEQ ID NO:

4) and RKRRKR (SEQ ID NO;

     5).

6. The nucleic acid sequence of claim 1, comprising a first DNA segment coding for the amino acids 1 through 720 of SEQ ID NO:

2 and a second DNA segment coding for the amino acids 1690 through 2332 of SEQ ID NO;

2, said segments being interconnected by at least one linker DNA segment coding for a PACE-furin or PACE furin-like cleavage site.

7. The nucleic acid sequence of claim 6, wherein said at least one PACE-furin or PACE furin-like cleavage site is selected from the group consisting of RRRR (SEQ ID NO:

4) and RKRRKR (SEQ ID NO;

     5).

8. The nucleic acid sequence of claim 1 further comprising 5′

and 3′

regulatory signals for expression of said nucleic acid sequence in transformed cells.

9. A vector comprising the nucleic acid sequence of claim 1.

10. An isolated modified human FVIII protein produced by expression of the nucleic acid of claim 1.

11. A method of treating hemophilia in a patient in need thereof, comprising:

a) providing a vector comprising the nucleic acid sequence of claim 1, operably linked to expression control elements; and

b) administering said vector to said patient under conditions that result in the expression of a modified human FVIII protein at a level that provides a therapeutic effect in said patient.

12. The method of claim 11, wherein said vector is selected from the group consisting of adenoviral-based vectors, adeno-associated viral vectors, retroviral vectors, and transposon-transposase vector systems.

13. A method of treating hemophilia in a patient in need thereof, comprising:

a) providing a variant FVIII polypeptide produced by the expression of the nucleic acid sequence of claim 1 in a biologically acceptable carrier; and

b) administering an effective amount of said polypeptide to said patient under conditions that result in a beneficial therapeutic effect in said patient.

14. The method of claim 13, wherein said biologically acceptable carrier further comprises a liposome.